Agroecology and sustainable food systems: Participatory research to improve food security among HIV-affected households in northern Malawi.
This article shares results from a long-term participatory agroecological research project in northern Malawi. Drawing upon a political ecology of health conceptual framework, the paper explores whether and how participatory agroecological farming can improve food security and nutrition among HIV-affected households. In-depth interviews were conducted with 27 farmers in HIV-affected households in the area near Ekwendeni Trading Centre in northern Malawi. The results show that participatory agroecological farming has a strong potential to meet the food, dietary, labour and income needs of HIV-affected households, whilst helping them to manage natural resources sustainably. As well, the findings reveal that place-based politics, especially gendered power imbalances, are imperative for understanding the human impacts of the HIV/AIDS epidemic. Overall, the study adds valuable insights into the literature on the human-environment dimensions of health. It demonstrates that the onset of disease can radically transform the social relations governing access to and control over resources (e.g., land, labour, and capital), and that these altered social relations in turn affect sustainable disease management. The conclusion highlights how the promotion of sustainable agroecology could help to partly address the socio-ecological challenges associated with HIV/AIDS.


Low Sensitivity of Abbreviated Tilt Table Testing for Diagnosing Postural Tachycardia Syndrome in Adults With ME/CFS.
Introduction: Orthostatic intolerance is common among individuals with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). In some ME/CFS case definitions, orthostatic intolerance is considered a core feature of the disorder. Some studies have employed tilt table tests lasting 2-5 min to diagnose one common form of orthostatic intolerance, postural tachycardia syndrome (POTS). We examined the diagnostic yield of abbreviated durations of tilt testing in adults meeting criteria for ME/CFS, and identified the proportion with POTS misdiagnosed using testing of <10 min. Methods: Eligible participants were consecutive individuals satisfying study criteria for ME/CFS and POTS evaluated at the Stichting CardioZorg (SCZ, Hoofddorp, NL) between November 2012 and August 2018. Individuals being treated with medications commonly used to manage orthostatic intolerance were excluded. Head-up tilt table testing involved 15 min of supine posture then 20 min at 70 degrees upright. Only the data from the first 10-min upright were used. POTS was defined as an increase in HR during a maximum of 10 min of upright tilt of at least 30 beats per minute (bpm), in the absence of either classical or delayed orthostatic hypotension. We measured the time until HR criteria for POTS were reached using survival curves, and compared survival curves between subgroups divided by age, sex, disease duration, and degree of hypocapnia during the test. Results: Of 627 individuals with ME/CFS evaluated during the study period, 155 met criteria for POTS. The median time to reaching HR criteria for POTS was 3 min. A two-minute tilt table test would miss 55% (95% CI, 48-63%) of those meeting POTS criteria over the course of 10 min upright. The median time to reaching HR criteria for POTS did not differ by sex, age, duration of ME/CFS, or hypocapnia during tilt. Conclusions: Abbreviated tilt table testing misses a substantial proportion of those ultimately diagnosed with POTS during a 10-min tilt table test, and should be abandoned for the clinical diagnosis and in epidemiologic studies designed to estimate the prevalence of POTS among those with ME/CFS.


Development of a list of high-risk operations for patients 65 years and older.
No consensus exists regarding the definition of high-risk surgery in older adults. An inclusive and precise definition of high-risk surgery may be useful for surgeons, patients, researchers, and hospitals. To develop a list of high-risk operations. Retrospective cohort study and modified Delphi procedure. The setting included all Pennsylvania acute care hospitals (Pennsylvania Health Care Cost Containment Council [PHC4] April 1, 2001, to December 31, 2007) and a nationally representative sample of US acute care hospitals (Nationwide Inpatient Sample [NIS], Healthcare Cost and Utilization Project, Agency for Healthcare Research and Quality January 1, 2001, to December 31, 2006). Patients included were those 65 years and older admitted to PHC4 hospitals and those 18 years and older admitted to NIS hospitals. We identified International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) procedure codes associated with at least 1% inpatient mortality in the PHC4. We used a modified Delphi procedure with 5 board-certified surgeons to further refine this list by excluding nonoperative procedures and operations that were unlikely to be the proximate cause of mortality and were instead a marker of critical illness (eg, tracheostomy). We then cross-validated this list of ICD-9-CM codes in the NIS. Modified Delphi procedure consensus of at least 4 of 5 panelists and proportion agreement in the NIS. Among 4,739,522 admissions of patients 65 years and older in the PHC4, a total of 2,569,589 involved a procedure, encompassing 2853 unique procedures. Of 1130 procedures associated with a crude inpatient mortality of at least 1%, 264 achieved consensus as high-risk operations by the modified Delphi procedure. The observed inpatient mortality in the NIS was at least 1% for 227 of 264 procedures (86%) in patients 65 years and older. The pooled inpatient mortality for these identified high-risk procedures performed on patients 65 years and older was double the pooled inpatient mortality for correspondingly identified high-risk operations for patients younger than 65 years (6% vs 3%). We developed a list of procedure codes to identify high-risk surgical procedures in claims data. This list of high-risk operations can be used to standardize the definition of high-risk surgery in quality and outcomes-based studies and to design targeted clinical interventions.


Comparison of EuroQol-5D and short form-6D utility scores in multiethnic Asian patients with psoriatic arthritis: a cross-sectional study.
To compare EuroQol-5D (EQ-5D) and Short Form-6D (SF-6D) utility scores in multiethnic Asian patients with psoriatic arthritis (PsA). Consecutive patients fulfilling the Classification Criteria for Psoriatic Arthritis attending a rheumatology outpatient clinic were recruited and completed the EQ-5D and SF-6D questionnaires. Comparisons were performed by score distribution, mean, median, and the Outcome Measures in Rheumatology filter: i.e., truth, discrimination, and feasibility. Eighty-six patients were enrolled (69 English-speaking and 17 Chinese-speaking; male:female ratio 0.91). The score distribution of SF-6D was normal, while that of EQ-5D was bimodal. A ceiling effect was observed in 20% of patients for EQ-5D and none for SF-6D. There were moderate correlations (Spearman's rho = 0.59, p < 0.0001) between the 2 scores, but poor agreements on scatterplot, intraclass correlation (ICC 0.43 and standardized ICC 0.21), and Bland-Altman plots. EQ-5D generated lower utility scores than SF-6D in the poorer health subgroup. SF-6D had stronger correlation with the general health status and other external measures of health; and it distinguished better between good and poor general health status, with better effect size and relative efficiency statistics. EQ-5D demonstrated higher patient acceptability. EQ-5D and SF-6D instruments generated different utility scores in PsA. SF-6D may be superior because of normal scaling distribution and the absence of ceiling and floor effects. SF-6D also had better construct validity and better discrimination of poor health status. More studies are required for cost-utility analysis in PsA.


Field use of D2 18O to measure energy expenditure of soldiers at different energy intakes.
To test the application of doubly labeled water under adverse field conditions, energy expenditures of 16 special operations soldiers were measured during a 28-day field training exercise. Subjects were matched by fat-free mass and divided equally between an ad libitum ready-to-eat meal diet and a 2,000 kcal/day lightweight ration. Subjects recorded intakes daily, and body composition was measured before and after the exercise. At the beginning of the study, subjects moved to a new northerly location and, therefore, a new water supply. To compensate for this, a group of soldiers who did not receive heavy water was followed to measure isotopic base-line changes. Energy expenditure by doubly labeled water was in agreement with intake/balance (3,400 +/- 260 vs. 3,230 +/- 520 kcal/day). The overall coefficient of variation of energy expenditure by doubly labeled water was half that of intake/balance (7.6 vs. 16.1%). The coefficient of variation of repeat measures with doubly labeled water was 7.3%. Energy expenditure of the ready-to-eat meal group, 3,540 +/- 180 kcal/day, was not significantly different from the lightweight ration group, 3,330 +/- 301 kcal/day. Doubly labeled water was valid under field conditions.


Evidence for ligand-dependent intramolecular folding of the AF-2 domain in vitamin D receptor-activated transcription and coactivator interaction.
A ligand-dependent transcriptional activation domain (AF-2) exists in region E of the nuclear receptors. This highly conserved domain may contact several coactivators that are putatively involved in nuclear receptor-mediated transcription. In this study, a panel of vitamin D receptor (VDR) AF-2 mutants was created to examine the importance of several conserved residues in VDR-activated transcription. Two AF-2 mutants (L417S and E420Q) exhibited normal ligand binding, heterodimerization with retinoid X receptor, and vitamin D-responsive element interaction, but they were transcriptionally inactive in a VDR-responsive reporter gene assay. All AF-2 mutations that abolished VDR-mediated transactivation also eliminated interactions between VDR and several putative coactivator proteins including suppressor of gal1 (SUG1), steroid hormone receptor coactivator-1 (SRC-1), or receptor interacting protein (RIP140), suggesting that coactivator interaction is important for AF-2-mediated transcription. In support of this concept, the minimal AF-2 domain [VDR(408-427)] fused to the gal4 DNA binding domain was sufficient to mediate transactivation as well as interaction with putative coactivators. Introducing the L417S and E420Q mutations into the minimal AF-2 domain abolished this autonomous transactivation and coactivator interactions. Finally, we demonstrate that the minimal AF-2 domain interacted with an AF-2 deletion mutant of the VDR in a 1,25-(OH)2D3-dependent manner, suggesting a ligand-induced intramolecular folding of the VDR AF-2 domain. The L417S mutant of this domain disrupted the interaction with VDR ligand-binding domain, while the E420Q mutant did not affect this interaction. These studies suggest that the conserved AF-2 motif may mediate transactivation through ligand-dependent intermolecular interaction with coactivators and through ligand-induced intramolecular contacts with the VDR ligand-binding domain itself.


Influence of vitamin B12 and cocultures on the growth of Dehalococcoides isolates in defined medium.
Bacteria belonging to the genus Dehalococcoides play a key role in the complete detoxification of chloroethenes as these organisms are the only microbes known to be capable of dechlorination beyond dichloroethenes to vinyl chloride (VC) and ethene. However, Dehalococcoides strains usually grow slowly with a doubling time of 1 to 2 days and have complex nutritional requirements. Here we describe the growth of Dehalococcoides ethenogenes 195 in a defined mineral salts medium, improved growth of strain 195 when the medium was amended with high concentrations of vitamin B(12), and a strategy for maintaining Dehalococcoides strains on lactate by growing them in consortia. Although strain 195 could grow in defined medium spiked with approximately 0.5 mM trichloroethene (TCE) and 0.001 mg/liter vitamin B(12), the TCE dechlorination and cellular growth rates doubled when the vitamin B(12) concentration was increased 25-fold to 0.025 mg/liter. In addition, the final ratios of ethene to VC increased when the higher vitamin concentration was used, which reflected the key role that cobalamin plays in dechlorination reactions. No further improvement in dechlorination or growth was observed when the vitamin B(12) concentration was increased to more than 0.025 mg/liter. In defined consortia containing strain 195 along with Desulfovibrio desulfuricans and/or Acetobacterium woodii and containing lactate as the electron donor, tetrachloroethene ( approximately 0.4 mM) was completely dechlorinated to VC and ethene and there was concomitant growth of Dehalococcoides cells. In the cultures that also contained D. desulfuricans and/or A. woodii, strain 195 cells grew to densities that were 1.5 times greater than the densities obtained when the isolate was grown alone. The ratio of ethene to VC was highest in the presence of A. woodii, an organism that generates cobalamin de novo during metabolism. These findings demonstrate that the growth of D. ethenogenes strain 195 in defined medium can be optimized by providing high concentrations of vitamin B(12) and that this strain can be grown to higher densities in cocultures with fermenters that convert lactate to generate the required hydrogen and acetate and that may enhance the availability of vitamin B(12).


Observational Study of the Effect of Patient Outreach on Return to Care: The Earlier the Better.
The burden of HIV remains heaviest in resource-limited settings, where problems of losses to care, silent transfers, gaps in care, and incomplete mortality ascertainment have been recognized. Patients in care at Academic Model Providing Access to Healthcare (AMPATH) clinics from 2001-2011 were included in this retrospective observational study. Patients missing an appointment were traced by trained staff; those found alive were counseled to return to care (RTC). Relative hazards of RTC were estimated among those having a true gap: missing a clinic appointment and confirmed as neither dead nor receiving care elsewhere. Sample-based multiple imputation accounted for missing vital status. Among 34,522 patients lost to clinic, 15,331 (44.4%) had a true gap per outreach, 2754 (8.0%) were deceased, and 837 (2.4%) had documented transfers. Of 15,600 (45.2%) remaining without active ascertainment, 8762 (56.2%) with later RTC were assumed to have a true gap. Adjusted cause-specific hazard ratios (aHRs) showed early outreach (a ≤8-day window, defined by grid-search approach) had twice the hazard for RTC vs. those without (aHR = 2.06; P < 0.001). HRs for RTC were lower the later the outreach effort after disengagement (aHR = 0.86 per unit increase in time; P < 0.001). Older age, female sex (vs. male), antiretroviral therapy use (vs. none), and HIV status disclosure (vs. none) were also associated with greater likelihood of RTC, and higher enrollment CD4 count with lower likelihood of RTC. Patient outreach efforts have a positive impact on patient RTC, regardless of when undertaken, but particularly soon after the patient misses an appointment.


Angoff's delta method revisited: improving DIF detection under small samples.
Most methods for detecting differential item functioning (DIF) are suitable when the sample sizes are sufficiently large to validate the null statistical distributions. There is no guarantee, however, that they will still perform adequately when there are few respondents in the focal group or in both the reference and the focal group. Angoff's delta plot is a potentially useful alternative for small-sample DIF investigation, but it suffers from an improper DIF flagging criterion. The purpose of this paper is to improve this classification rule under mild statistical assumptions. This improvement yields a modified delta plot with an adjusted DIF flagging criterion for small samples. A simulation study was conducted to compare the modified delta plot with both the classical delta plot approach and the Mantel-Haenszel method. It is concluded that the modified delta plot is consistently less conservative and more powerful than the usual delta plot, and is also less conservative and more powerful than the Mantel-Haenszel method as long as at least one group of respondents is small.


Why does a trait evolve multiple times within a clade? Repeated evolution of snakelike body form in squamate reptiles.
Why does a trait evolve repeatedly within a clade? When examining the evolution of a trait, evolutionary biologists typically focus on the selective advantages it may confer and the genetic and developmental mechanisms that allow it to vary. Although these factors may be necessary to explain why a trait evolves in a particular instance, they may not be sufficient to explain phylogenetic patterns of repeated evolution or conservatism. Instead, other factors may also be important, such as biogeography and competitive interactions. In squamate reptiles (lizards and snakes) a dramatic transition in body form has occurred repeatedly, from a fully limbed, lizardlike body form to a limb-reduced, elongate, snakelike body form. We analyze this trait in a phylogenetic and biogeographic context to address why this transition occurred so frequently. We included 261 species for which morphometric data and molecular phylogenetic information were available. Among the included species, snakelike body form has evolved about 25 times. Most lineages of snakelike squamates belong to one of two "ecomorphs," either short-tailed burrowers or long-tailed surface dwellers. The repeated origins of snakelike squamates appear to be associated with the in situ evolution of these two ecomorphs on different continental regions (including multiple origins of the burrowing morph within most continents), with very little dispersal of most limb-reduced lineages between continental regions. Overall, the number of repeated origins of snakelike morphology seems to depend on large-scale biogeographic patterns and community ecology, in addition to more traditional explanations (e.g., selection, development).


Hierarchical recognition scheme for human facial expression recognition systems.
Over the last decade, human facial expressions recognition (FER) has emerged as an important research area. Several factors make FER a challenging research problem. These include varying light conditions in training and test images; need for automatic and accurate face detection before feature extraction; and high similarity among different expressions that makes it difficult to distinguish these expressions with a high accuracy. This work implements a hierarchical linear discriminant analysis-based facial expressions recognition (HL-FER) system to tackle these problems. Unlike the previous systems, the HL-FER uses a pre-processing step to eliminate light effects, incorporates a new automatic face detection scheme, employs methods to extract both global and local features, and utilizes a HL-FER to overcome the problem of high similarity among different expressions. Unlike most of the previous works that were evaluated using a single dataset, the performance of the HL-FER is assessed using three publicly available datasets under three different experimental settings: n-fold cross validation based on subjects for each dataset separately; n-fold cross validation rule based on datasets; and, finally, a last set of experiments to assess the effectiveness of each module of the HL-FER separately. Weighted average recognition accuracy of 98.7% across three different datasets, using three classifiers, indicates the success of employing the HL-FER for human FER.


Calculating accurate proton chemical shifts of organic molecules with density functional methods and modest basis sets.
The purpose of this paper is to convince practitioners of (1)H NMR spectroscopy to consider simple quantum chemical calculations as a viable option to aid them in the assignment of their spectra. To this end, it is demonstrated, on a test set of 80 conformationally stable molecules of various kinds carrying different functional groups, that, in contrast to what is claimed in the literature, large basis sets are not needed to obtain rather accurate predictions of (1)H NMR chemical shifts by quantum chemical calculations. On the other hand, modeling the solvent by an SCRF-type calculation may improve certain predictions significantly. The best accuracy/cost ratio is provided by GIAO calculations in chloroform as a solvent with the specially parametrized WP04 functional of Cramer et al. using the cc-pVDZ or 6-31G** basis set, closely followed by similar calculations with the ubiquitious B3LYP functional (both predict (1)H chemical shifts with an average deviation of ca. 0.12 ppm, if the results are scaled linearly). A slightly higher accuracy can be attained by adding diffuse functions to the basis set, but going to the triple-zeta basis sets which have invariably been used hitherto in calculations of chemical shifts does not lead to any improvement. The popular increment schemes such as those implemented in the ChemDraw or ACD programs do not do nearly as well and are often incapable of correctly distinguishing stereoisomers.


Examining the pathogenesis of breast cancer using a novel agent-based model of mammary ductal epithelium dynamics.
The study of the pathogenesis of breast cancer is challenged by the long time-course of the disease process and the multi-factorial nature of generating oncogenic insults. The characterization of the longitudinal pathogenesis of malignant transformation from baseline normal breast duct epithelial dynamics may provide vital insight into the cascading systems failure that leads to breast cancer. To this end, extensive information on the baseline behavior of normal mammary epithelium and breast cancer oncogenesis was integrated into a computational model termed the Ductal Epithelium Agent-Based Model (DEABM). The DEABM is composed of computational agents that behave according to rules established from published cellular and molecular mechanisms concerning breast duct epithelial dynamics and oncogenesis. The DEABM implements DNA damage and repair, cell division, genetic inheritance and simulates the local tissue environment with hormone excretion and receptor signaling. Unrepaired DNA damage impacts the integrity of the genome within individual cells, including a set of eight representative oncogenes and tumor suppressors previously implicated in breast cancer, with subsequent consequences on successive generations of cells. The DEABM reproduced cellular population dynamics seen during the menstrual cycle and pregnancy, and demonstrated the oncogenic effect of known genetic factors associated with breast cancer, namely TP53 and Myc, in simulations spanning ∼40 years of simulated time. Simulations comparing normal to BRCA1-mutant breast tissue demonstrated rates of invasive cancer development similar to published epidemiologic data with respect to both cumulative incidence over time and estrogen-receptor status. Investigation of the modeling of ERα-positive (ER+) tumorigenesis led to a novel hypothesis implicating the transcription factor and tumor suppressor RUNX3. These data suggest that the DEABM can serve as a potentially valuable framework to augment the traditional investigatory workflow for future hypothesis generation and testing of the mechanisms of breast cancer oncogenesis.


A Drosophila protein-interaction map centered on cell-cycle regulators.
Maps depicting binary interactions between proteins can be powerful starting points for understanding biological systems. A proven technology for generating such maps is high-throughput yeast two-hybrid screening. In the most extensive screen to date, a Gal4-based two-hybrid system was used recently to detect over 20,000 interactions among Drosophila proteins. Although these data are a valuable resource for insights into protein networks, they cover only a fraction of the expected number of interactions. To complement the Gal4-based interaction data, we used the same set of Drosophila open reading frames to construct arrays for a LexA-based two-hybrid system. We screened the arrays using a novel pooled mating approach, initially focusing on proteins related to cell-cycle regulators. We detected 1,814 reproducible interactions among 488 proteins. The map includes a large number of novel interactions with potential biological significance. Informative regions of the map could be highlighted by searching for paralogous interactions and by clustering proteins on the basis of their interaction profiles. Surprisingly, only 28 interactions were found in common between the LexA- and Gal4-based screens, even though they had similar rates of true positives. The substantial number of new interactions discovered here supports the conclusion that previous interaction mapping studies were far from complete and that many more interactions remain to be found. Our results indicate that different two-hybrid systems and screening approaches applied to the same proteome can generate more comprehensive datasets with more cross-validated interactions. The cell-cycle map provides a guide for further defining important regulatory networks in Drosophila and other organisms.


Magnetoresistive polyaniline-silicon carbide metacomposites: plasma frequency determination and high magnetic field sensitivity.
The Drude model modified by Debye relaxation time was introduced to determine the plasma frequency (ωp) in the surface initiated polymerization (SIP) synthesized β-silicon carbide (β-SiC)/polyaniline (PANI) metacomposites. The calculated plasma frequency for these metacomposites with different loadings of β-SiC nanoparticles was ranging from 6.11 × 10(4) to 1.53 × 10(5) rad s(-1). The relationship between the negative permittivity and plasma frequency indicates the existence of switching frequency, at which the permittivity was changed from negative to positive. More interestingly, the synthesized non-magnetic metacomposites, observed to follow the 3-dimensional (3-D) Mott variable range hopping (VRH) electrical conduction mechanism, demonstrated high positive magnetoresistance (MR) values of up to 57.48% and high MR sensitivity at low magnetic field regimes.


Novel type V staphylococcal cassette chromosome mec driven by a novel cassette chromosome recombinase, ccrC.
Staphylococcal cassette chromosome mec (SCCmec) is a mobile genetic element composed of the mec gene complex, which encodes methicillin resistance, and the ccr gene complex, which encodes the recombinases responsible for its mobility. The mec gene complex has been classified into four classes, and the ccr gene complex has been classified into three allotypes. Different combinations of mec gene complex classes and ccr gene complex types have so far defined four types of SCCmec elements. Now we introduce the fifth allotype of SCCmec, which was found on the chromosome of a community-acquired methicillin-resistant Staphylococcus aureus strain (strain WIS [WBG8318]) isolated in Australia. The element shared the same chromosomal integration site with the four extant types of SCCmec and the characteristic nucleotide sequences at the chromosome-SCCmec junction regions. The novel SCCmec carried mecA bracketed by IS431 (IS431-mecA-DeltamecR1-IS431), which is designated the class C2 mec gene complex; and instead of ccrA and ccrB genes, it carried a single copy of a gene homologue that encoded cassette chromosome recombinase. Since the open reading frame (ORF) was found to encode an enzyme which catalyzes the precise excision as well as site- and orientation-specific integration of the element, we designated the ORF cassette chromosome recombinase C (ccrC), and we designated the element type V SCCmec. Type V SCCmec is a small SCCmec element (28 kb) and does not carry any antibiotic resistance genes besides mecA. Unlike the extant SCCmec types, it carries a set of foreign genes encoding a restriction-modification system that might play a role in the stabilization of the element on the chromosome.


Missing: critical and skeptical perspectives on comprehensive soldier fitness.
The author read with interest, and concern, the January 2011 issue of the American Psychologist. The Special Issue on Comprehensive Soldier Fitness addressed a hugely consequential national issue and offered a vision for psychological resilience along with an elaborate set of supporting articles, concluding with some comments on "Objections" to psychologists working with the military. While a bold vision was offered and a game plan presented, no data or evidence are yet available (it is far too early in the process), and that should concern us all. The author hopes the American Psychological Association (APA) and the American Psychologist offer skeptical and critical views in future issues on the issue of the psychological health of our military and as evidence emerges regarding the Comprehensive Soldier Fitness program.


Blind deconvolution in optical diffusion tomography.
We use blind deconvolution methods in optical diffusion tomography to reconstruct images of objects imbedded in or located behind turbid media from continuous-wave measurements of the scattered light transmitted through the media. In particular, we use a blind deconvolution imaging algorithm to determine both a deblurred image of the object and the depth of the object inside the turbid medium. Preliminary results indicate that blind deconvolution produces better reconstructions than can be obtained using backpropagation techniques. Moreover, it does so without requiring prior knowledge of the characteristics of the turbid medium or of what the blur-free target should look like: important advances over backpropagation.


Can virtual nature improve patient experiences and memories of dental treatment? A study protocol for a randomized controlled trial.
Dental anxiety and anxiety-related avoidance of dental care create significant problems for patients and the dental profession. Distraction interventions are used in daily medical practice to help patients cope with unpleasant procedures. There is evidence that exposure to natural scenery is beneficial for patients and that the use of virtual reality (VR) distraction is more effective than other distraction interventions, such as watching television. The main aim of this randomized controlled trial is to determine whether the use of VR during dental treatment can improve the overall dental experience and recollections of treatment for patients, breaking the negative cycle of memories of anxiety leading to further anxiety, and avoidance of future dental appointments. Additionally, the aim is to test whether VR benefits dental patients with all levels of dental anxiety or whether it could be especially beneficial for patients suffering from higher levels of dental anxiety. The third aim is to test whether the content of the VR distraction can make a difference for its effectiveness by comparing two types of virtual environments, a natural environment and an urban environment. The effectiveness of VR distraction will be examined in patients 18 years or older who are scheduled to undergo dental treatment for fillings and/or extractions, with a maximum length of 30 minutes. Patients will be randomly allocated into one of three groups. The first group will be exposed to a VR of a natural environment. The second group will be exposed to a VR of an urban environment. A third group consists of patients who receive standard care (control group). Primary outcomes relate to patients' memories of the dental treatment one week after treatment: (a) remembered pain, (b) intrusive thoughts and (c) vividness of memories. Other measures of interest are the dental experience, the treatment experience and the VR experience. Current Controlled Trials ISRCTN41442806.


Predictors for length of hospital stay in patients with community-acquired pneumonia: results from a Swiss multicenter study.
Length of hospital stay (LOS) in patients with community-acquired pneumonia (CAP) is variable and directly related to medical costs. Accurate estimation of LOS on admission and during follow-up may result in earlier and more efficient discharge strategies. This is a prospective multicenter study including patients in emergency departments of 6 tertiary care hospitals in Switzerland between October 2006 and March 2008. Medical history, clinical data at presentation and health care insurance class were collected. We calculated univariate and multivariate cox regression models to assess the association of different characteristics with LOS. In a split sample analysis, we created two LOS prediction rules, first including only admission data, and second including also additional inpatient information. The mean LOS in the 875 included CAP patients was 9.8 days (95%CI 9.3-10.4). Older age, respiratory rate >20 pm, nursing home residence, chronic pulmonary disease, diabetes, multilobar CAP and the pneumonia severity index class were independently associated with longer LOS in the admission prediction model. When also considering follow-up information, low albumin levels, ICU transfer and development of CAP-associated complications were additional independent risk factors for prolonged LOS. Both weighted clinical prediction rules based on these factors showed a high separation of patients in Kaplan Meier Curves (p logrank <0.001 and <0.001) and a good calibration when comparing predicted and observed results. Within this study we identified different baseline and follow-up characteristics to be strong and independent predictors for LOS. If validated in future studies, these factors may help to optimize discharge strategies and thus shorten LOS in CAP patients.


Anal screening cytology.
This issue of CytoJournal contains an article on screening for anal intraepithelial neoplasia in high-risk male patients. This accompanying Editorial focuses on current understanding of this relatively new disease entity, with insights as to the potential role of screening cytopathology in the epidemiology, pathophysiology and clinical management of this HIV and HPV related anal lesion, which predominates in male patients living long-term with AIDS. Mention is made of techniques of obtaining samples, methods of preparation, and morphologic classification. Issues of anoscopic confirmation, as well as topical and surgical management are emphasized. The similarity of initial experiences in anal screening to problems encountered early in cervical cancer screening programs several decades ago, are highlighted.


A dynamic model of pneumococcal infection in the United States: implications for prevention through vaccination.
Universal infant vaccination with the 7-valent pneumococcal conjugate vaccine (PCV7) has nearly eliminated PCV7-serotype invasive pneumococcal disease (IPD) in young U.S. children, but has been accompanied by increases in the incidence of serotype 19A IPD. Because antibiotic-non-susceptible 19A has increased more than antibiotic-susceptible 19A, antibiotic selection pressure could be contributing to this trend. We developed a dynamic compartmental transmission model of pneumococcus to better understand the causes of this rise and to estimate the impact of vaccines or changes in antibiotic use on future IPD incidence in the U.S. in <2 year-olds. The model predicted that with current practices, serotype 19A IPD incidence will plateau at about the 2007 level over the next few years. The model suggests that antibiotic usage played a major role in the rise in antibiotic-non-susceptible 19A IPD, with a lesser contribution from PCV7 vaccination. However, hypothetical large decreases in antibiotic use starting in 2008 are predicted to yield only gradual decreases in antibiotic-non-susceptible 19A IPD. On the other hand, vaccines with modest (20%) effectiveness against 19A (or 6A or PCV7-serotypes) carriage are predicted to substantially (by 80%) decrease the incidence of IPD caused by those serotypes within 10 years of implementation. Our findings highlight that vaccine effects on colonization are key to their overall benefits. In addition, serotype changes following vaccine introduction may have multifactorial origins, with antibiotic use an important factor for resistant strains such as 19A.


Treponema pallidum hemagglutination assay seroreactivity among healthy Indian donors and its association with other transfusion transmitted diseases.
The aim of the present study was to determine the prevalence of syphilis infection by Treponema pallidum hemagglutination assay (TPHA) among blood donors in Delhi and to study their correlation with other markers of transfusion transmitted infections such as hepatitis C virus (HCV), human immunodeficiency virus (HIV) and hepatitis B surface antigen (HBsAg) so as to establish the utility of TPHA over and above venereal diseases research laboratory test (VDRL), not only as a marker for testing T. pallidum infection, but also as a marker of high risk behavior. This prospective study was carried out in the Regional Blood Transfusion Centre, Lady Hardinge Medical College and associated Sucheta Kriplani Hospital, New Delhi for a period of 2 years. Donated blood was screened for TPHA seroreactivity along with screening for anti HIV I and II, anti-HCV, HBsAg by third generation enzyme-linked immunosorbent assay test. A total of 8082 serum samples of blood donors were collected from healthy blood donors in our blood bank. They were classified into two groups- test group and control group based on TPHA positivity. The co-occurrence of HBsAg, HIV and HCV infection were determined in TPHA positive blood donors (test group) in comparison with TPHA negative blood donors (control group). We found the TPHA seroreactivity to be 4.4% in Delhi's blood donors. Nearly 8.2% (663/8082) of the donated blood had serological evidence of infection by at least one pathogen (syphilis/HIV/hepatitis B virus/HCV) and 6.63% (44/663) donors with positive serology had multiple infections (two or more). Quadruple infection was seen in one donor, triple infection was seen in three donors and double infection was seen in 40 donors. Prevalence of HIV seroreactivity was found to be statistically significant and HCV seroreactivity statistically insignificant in TPHA positive group in comparison to TPHA negative group. In our study, the TPHA seropositivity correlated with higher HIV and HCV seropositivity and the same correlation has been observed by several other studies also. In view of these observations, we propose that testing for syphilis by more sensitive and specific treponemal markers like TPHA rather than VDRL, rapid plasma reagin tests; as TPHA also has the added advantage of picking up all the high risk donors, whereas, VDRL picks up only currently infected donors. Moreover, TPHA should be continued as a marker of high risk behavior especially in high prevalence areas like India where we don't have universal access to markers like nucleic acid amplification technique.


Longitudinal cohort study of depression, post-traumatic stress, and alcohol use in South African women who attend alcohol serving venues.
In South Africa, alcohol use poses a public health burden. Hazardous alcohol use often co-occurs with psychological distress (e.g., depression and post-traumatic stress). However, the majority of the research establishing the relationship between alcohol use and psychological distress has been cross-sectional, so the nature of co-occurring changes in psychological distress and alcohol use over time is not well characterized. The objective of this study is to examine the longitudinal relationship between psychological distress and alcohol use among South African women who attend alcohol serving venues. Four waves of data were collected over the course of a year from 560 women in a Cape Town township who attended drinking venues. At each assessment wave, participants reported depressive symptoms, post-traumatic stress symptoms, and alcohol use. Multilevel growth models were used to: 1) assess the patterns of alcohol use; 2) examine how depressive symptoms uniquely, post-traumatic stress symptoms uniquely, and depressive and post-traumatic stress symptoms together were associated with alcohol use; and 3) characterize the within person and between person associations of depressive symptoms and post-traumatic stress symptoms with alcohol use. Women reported high levels of alcohol use throughout the study period, which declined slightly over time. Post-traumatic stress symptoms were highly correlated with depressive symptoms. Modeled separately, both within person and between person depressive and post-traumatic stress symptoms were uniquely associated with alcohol use. When modeled together, significant between person effects indicated that women who typically have more post-traumatic stress symptoms, when controlling for depressive symptoms, are at risk for increased alcohol use; however, women with more depressive symptoms, controlling for post-traumatic stress symptoms, do not have differential risk for alcohol use. Significant within person effects indicated an interaction between depressive and post-traumatic stress symptoms; women reported more alcohol use than usual at times when they had higher post-traumatic stress symptoms, and this increase in alcohol use was further exacerbated for women who also had higher depressive symptoms than usual. These findings suggest that interventions targeting post-traumatic stress, especially when post-traumatic stress is comorbid with depression, may reduce alcohol use among South African women who drink.


Human neural stem cell-derived cultures in three-dimensional substrates form spontaneously functional neuronal networks.
Differentiated human neural stem cells were cultured in an inert three-dimensional (3D) scaffold and, unlike two-dimensional (2D) but otherwise comparable monolayer cultures, formed spontaneously active, functional neuronal networks that responded reproducibly and predictably to conventional pharmacological treatments to reveal functional, glutamatergic synapses. Immunocytochemical and electron microscopy analysis revealed a neuronal and glial population, where markers of neuronal maturity were observed in the former. Oligonucleotide microarray analysis revealed substantial differences in gene expression conferred by culturing in a 3D vs a 2D environment. Notable and numerous differences were seen in genes coding for neuronal function, the extracellular matrix and cytoskeleton. In addition to producing functional networks, differentiated human neural stem cells grown in inert scaffolds offer several significant advantages over conventional 2D monolayers. These advantages include cost savings and improved physiological relevance, which make them better suited for use in the pharmacological and toxicological assays required for development of stem cell-based treatments and the reduction of animal use in medical research. Copyright © 2015 John Wiley & Sons, Ltd.


Strong resonance and a highly compact longperiod grating in a large-mode-area photonic crystal fiber.
A strong resonance and extremely short length long-period grating (LPG) has been fabricated in a large-mode-area photonic crystal fiber (PCF) by use of a CO2 laser heat source. We believe that such a longperiod grating in pure silica PCF is the first example of a point-by-point technique. The fabrication method is simple and repeatable. The resulting LPG has been developed with eight periodic collapses within a 2.8-mmlong period of the fiber, which gives the strong resonance of core-cladding mode coupling. The lowest mode of LP01 is at a 1529.2-nm wavelength with a full width at half-maximum of ~0.7 nm and a resonance strength of -31.5 dB. The principal advantages of this LPG are that (1) it is temperature insensitive and stable, (2) the device is compact when it is packaged, and (3) it provides practical, low-cost all-fiber filters and PCF-based devices for optical fiber communications and sensing systems.


Factors influencing students' decisions to participate in a short "dissection experience" within a systemic anatomy course.
Changes in medical education have affected both curriculum design and delivery. Many medical schools now use integrated curricula and a systemic approach, with reduced hours of anatomy teaching. While learning anatomy via dissection is invaluable in educational, professional, and personal development, it is time intensive and supports a regional approach to learning anatomy; the use of prosections has replaced dissection as the main teaching method in many medical schools. In our graduate-entry medical degree, we use an integrated curriculum, with prosections to teach anatomy systemically. However, to not exclude dissection completely, and to expose students to its additional and unique benefits, we implemented a short "Dissection Experience" at the beginning of Year 2. Students attended three two-hour anatomy sessions and participated in dissection of the clinically relevant areas of the cubital fossa, femoral triangle, and infraclavicular region. This activity was voluntary and we retrospectively surveyed all students to ascertain factors influencing their decision of whether to participate in this activity, and to obtain feedback from those students who did participate. The main reasons students did not participate were previous dissection experience and time constraints. The reasons most strongly affecting students' decisions to participate related to experience (lack of previous or new) and new skill. Students' responses as to the most beneficial component of the dissection experience were based around practical skills, anatomical education, the learning process, and the body donors. We report here on the benefits and practicalities of including a short dissection experience in a systemic, prosection-based anatomy course.


Creation of a comprehensive training and career development approach to increase the number of neurosurgeons supported by National Institutes of Health funding.
To increase the number of independent National Institutes of Health (NIH)-funded neurosurgeons and to enhance neurosurgery research, the National Institute of Neurological Disorders and Stroke (NINDS) developed two national comprehensive programs (R25 [established 2009] for residents/fellows and K12 [2013] for early-career neurosurgical faculty) in consultation with neurosurgical leaders and academic departments to support in-training and early-career neurosurgeons. The authors assessed the effectiveness of these NINDS-initiated programs to increase the number of independent NIH-funded neurosurgeon-scientists and grow NIH neurosurgery research funding. NIH funding data for faculty and clinical department funding were derived from the NIH, academic departments, and Blue Ridge Institute of Medical Research databases from 2006 to 2019. Between 2009 and 2019, the NINDS R25 funded 87 neurosurgical residents. Fifty-three (61%) have completed the award and training, and 39 (74%) are in academic practice. Compared to neurosurgeons who did not receive R25 funding, R25 awardees were twice as successful (64% vs 31%) in obtaining K-series awards and received the K-series award in a significantly shorter period of time after training (25.2 ± 10.1 months vs 53.9 ± 23.0 months; p < 0.004). Between 2013 and 2019, the NINDS K12 has supported 19 neurosurgeons. Thirteen (68%) have finished their K12 support and all (100%) have applied for federal funding. Eleven (85%) have obtained major individual NIH grant support. Since the establishment of these two programs, the number of unique neurosurgeons supported by either individual (R01 or DP-series) or collaborative (U- or P-series) NIH grants increased from 36 to 82 (a 2.3-fold increase). Overall, NIH funding to clinical neurological surgery departments between 2006 and 2019 increased from $66.9 million to $157.3 million (a 2.2-fold increase). Targeted research education and career development programs initiated by the NINDS led to a rapid and dramatic increase in the number of NIH-funded neurosurgeon-scientists and total NIH neurosurgery department funding.


Acute symptomatic hypoglycaemia mimicking ischaemic stroke on imaging: a systemic review.
Acute symptomatic hypoglycaemia is a differential diagnosis in patients presenting with stroke-like neurological impairment, but few textbooks describe the full brain imaging appearances. We systematically reviewed the literature to identify how often hypoglycaemia may mimic ischaemic stroke on imaging, common patterns and relationships with hypoglycaemia severity, duration, clinical outcome and add two new cases. We searched EMBASE and Medline databases for papers reporting imaging in adults with symptomatic hypoglycaemia. We analysed the clinical presentation, outcome, brain imaging findings, duration and severity of hypoglycaemia, time course of lesion appearance, including two new cases. We found 42 papers describing computed tomography or magnetic resonance imaging in 65 patients, plus our two cases with symptomatic hypoglycaemia. Imaging abnormalities on computed tomography and magnetic resonance were uni or bilateral, cortical or sub-cortical. Thirteen (20%) mimicked cortical or lacunar stroke. Acute lesions had restricted diffusion on magnetic resonance or low attenuation on computed tomography, plus swelling; older lesions showed focal atrophy or disappeared, as with ischaemic stroke. The association between the depth or duration of hypoglycaemia, the severity or extent of neurological deficit, and the imaging abnormalities, was weak. Imaging abnormalities in patients with hypoglycaemia are uncommon but very variable, weakly associated with neurological deficit, and about a fifth mimic acute ischaemic stroke. Blood glucose testing should be routine in all patients with acute neurological impairment and hypoglycaemia should be included in the differential diagnosis of imaging appearances in patients presenting with acute stroke.


Prediction of survival time in advanced cancer: a prognostic scale for Chinese patients.
This study reports the development of a simple Chinese Prognostic Scale (ChPS) for predicting survival in advanced cancer patients. Data relating to 1,019 advanced cancer patients referred to a palliative home care service were retrospectively analyzed. The records were divided into two sets using stratified random sampling: 80% as a "training set" for developing the scale and 20% as a "testing set" for validating it. Demographic data, symptoms/signs, Karnofsky Performance Status (KPS), quality of life (QOL), and survival time were statistically analyzed to create the scale. In the training set, a total of 10 prognostic factors were determined: weight loss, nausea, dysphagia, dyspnea, edema, cachexia, dehydration, gender, KPS, and QOL. The ChPS score was calculated for each case by summing the partial scores of prognostic factors, ranging from 0 (no altered variables) to 124 (maximal altered variables). The score for a cutoff point of three months' survival was 28 (95% confidence interval: 26.6, 28.9). When scores were more than 28, survival appeared to be usually less than three months. The accuracy rate was 69.4% in the training set and 65.4% in the testing set. In conclusion, it is possible with this prognostic scale to guide physicians in predicting more accurately the likely survival time of Chinese cancer patients, and to help policy makers in establishing appropriate referral for hospice care.


An immunogenomic stratification of colorectal cancer: Implications for development of targeted immunotherapy.
Although tumor infiltrating lymphocyte (TIL) density is prognostic and predictive in colorectal cancer (CRC), the impact of tumor genetics upon colorectal immunobiology is unclear. Identification of genetic factors that influence the tumor immunophenotype is essential to improve the effectiveness of stratified immunotherapy approaches. We carried out a bioinformatics analysis of CRC data in The Cancer Genome Atlas (TCGA) involving two-dimensional hierarchical clustering to define an immune signature that we used to characterize the immune response across key patient groups. An immune signature termed The Co-ordinate Immune Response Cluster (CIRC) comprising 28 genes was coordinately regulated across the patient population. Four patient groups were delineated on the basis of cluster expression. Group A, which was heavily enriched for patients with microsatellite instability (MSI-H) and POL mutations, exhibited high CIRC expression, including the presence of several inhibitory molecules: CTLA4, PDL1, PDL2, LAG3, and TIM3. In contrast, RAS mutation was enriched in patient groups with lower CIRC expression. This work links the genetics and immunobiology of colorectal tumorigenesis, with implications for the development of stratified immunotherapeutic approaches. Microsatellite instability and POL mutations are linked with high mutational burden and high immune infiltration, but the coordinate expression of inhibitory pathways observed suggests combination checkpoint blockade therapy may be required to improve efficacy. In contrast, RAS mutant tumors predict for a relatively poor immune infiltration and low inhibitory molecule expression. In this setting, checkpoint blockade may be less efficacious, highlighting a requirement for novel strategies in this patient group.


Venous reconstruction for iliofemoral venous occlusion facilitated by temporary arteriovenous shunt. Long-term results in nine patients.
In nine patients with iliofemoral venous occlusion, venous reconstructions using a temporary arteriovenous shunt were performed by open thromboendvenectomy with autogenous vein patch angioplasty in four, expanded polytetrafluoroethylene (ePTFE) bypass grafts (including two with external ring-supported ePTFE) in four, and Palma's procedure in one patient. There was an adequate function in the reconstructed venous segments in two of four who underwent thromboendvenectomy and in all four with ePTFE bypass grafting for nine months to 13 years after surgery. In those with a temporary arteriovenous shunt, prepared to maintain patency of the reconstructed venous segments, blood flow through the shunt exceeded 100 mL/min, determined by an electromagnetic flowmeter. Postoperative shunt closure was readily facilitated, using a looping technique and a 2-0 nylon. The increased blood flow through the graft made feasible by the temporary arteriovenous shunt enhanced the patency of the reconstructed venous graft and hence there was an improvement in the affected limb.


Expertise Across Disciplines: Establishing Common Ground in Interdisciplinary Disaster Research Teams.
Hazards and disasters arise from interactions between environmental and social processes, so interdisciplinary research is crucial in understanding and effectively managing them. Despite support and encouragement from funding agencies, universities, and journals and growing interest from researchers, interdisciplinary disaster research teams face significant obstacles, such as the difficulty of establishing effective communication and understanding across disciplines. Better understanding of interdisciplinary teamwork can also have important practical benefits for operational disaster planning and response. Social studies of science distinguish different kinds of expertise and different modes of communication. Understanding these differences can help interdisciplinary research teams communicate more clearly and work together more effectively. The primary role of a researcher is in contributory expertise (the ability to make original contributions to a discipline); but interactional expertise in other disciplines (the ability to understand their literature and communicate with their practitioners) can play an important role in interdisciplinary collaborations. Developing interactional expertise requires time and effort, which can be challenging for a busy researcher, and also requires a foundation of trust and communication among team members. Three distinct aspects of communication play important roles in effective interdisciplinary communication: dialects, metaphors, and articulation. There are different ways to develop interactional expertise and effective communication, so researchers can pursue approaches that suit their circumstances. It will be important for future research on interdisciplinary disaster research to identify best practices for building trust, facilitating communication, and developing interactional expertise.


Cognitive remediation therapy in schizophrenia: cost-effectiveness analysis.
There is a lack of evidence on the cost-effectiveness of cognitive remediation therapy (CRT). Randomised controlled trial comparing usual care plus CRT with usual care alone. Participants had a diagnosis of schizophrenia and cognitive and social functioning difficulties. Health/social care and societal costs were estimated at 14 weeks (time 2) and 40 weeks (time 3) after randomisation. The outcome, proportion of participants improving their working memory since baseline, was combined with costs to explore cost-effectiveness. 85 participants were recruited. There were no differences in total health/social care or societal costs between the two groups at either time 2 or time 3. An additional 21% of participants in the CRT group improved their working memory at both follow-ups. When placing these cost and outcomes in hypothetical scenarios concerning how much policy-makers would pay for another 1% of participants improving their working memory, there was more than an 80% chance that CRT would be cost-effective compared to usual care; at time 3, the likelihood of cost-effectiveness peaked at 30% even for investments up to pound 5000. CRT can improve memory among people with schizophrenia and cognitive deficits at no additional cost. Although cost-effective in the short term, CRT may have limited potential to save costs in the medium term because it could increase take up of services. This could confer important longer term benefits for the patient group examined here, in terms of improved social functioning and less reliance on services. This can only be ascertained through longer follow-up.


Asian-Pacific clinical practice guidelines on the management of hepatitis B: a 2015 update.
Worldwide, some 240 million people have chronic hepatitis B virus (HBV), with the highest rates of infection in Africa and Asia. Our understanding of the natural history of HBV infection and the potential for therapy of the resultant disease is continuously improving. New data have become available since the previous APASL guidelines for management of HBV infection were published in 2012. The objective of this manuscript is to update the recommendations for the optimal management of chronic HBV infection. The 2015 guidelines were developed by a panel of Asian experts chosen by the APASL. The clinical practice guidelines are based on evidence from existing publications or, if evidence was unavailable, on the experts' personal experience and opinion after deliberations. Manuscripts and abstracts of important meetings published through January 2015 have been evaluated. This guideline covers the full spectrum of care of patients infected with hepatitis B, including new terminology, natural history, screening, vaccination, counseling, diagnosis, assessment of the stage of liver disease, the indications, timing, choice and duration of single or combination of antiviral drugs, screening for HCC, management in special situations like childhood, pregnancy, coinfections, renal impairment and pre- and post-liver transplant, and policy guidelines. However, areas of uncertainty still exist, and clinicians, patients, and public health authorities must therefore continue to make choices on the basis of the evolving evidence. The final clinical practice guidelines and recommendations are presented here, along with the relevant background information.


The effect of heavy social drinking on recall and event-related potentials.
It has been suggested that alcohol-related brain impairments progress on a continuum from subtle deficits in social drinkers through to end-state Korsakoff syndrome. The aim of the present study was to investigate whether heavy levels of social drinking have an adverse effect on sober cognitive functioning in young nonalcoholic adults. Event-related potentials (ERPs) were recorded from heavy ( > 200 gm/week) and light ( < 20 gm/week) male social drinkers under the effects of a pharmacological challenge (lorazepam-ATIVAN) and a placebo. The heavy (n = 14) and light (n = 14) social drinkers completed a free-recall task with a rare word probability of .30. Analysis of the P300 component of the ERP to recalled and not-recalled words found that P300 amplitude in heavy social drinkers (HSDs) was reduced following placebo compared to the light social drinkers (LSDs). Lorazepam produced a distinctive pattern of anterograde memory deficits in both groups and reduced P300 amplitude to rare words in the LSDs compared to both the placebo treatment and the HSDs. The differences in central nervous system functioning evident between HSDs and LSDs were reflected in ERP deviations in both the presence and absence of lorazepam. Even though no definite statements can be made because of the small sample size, the results appear to indicate that HSDs have an impairment in their information-processing ability in the absence of an acute inhibitory agent (e.g., alcohol or a benzodiazepine) as well as a differential response to the depressant effects of lorazepam.


Electroanatomical mapping based on discrimination of electrograms clusters for localization of critical sites in atrial fibrillation.
Locating critical sites on the atrial surface during AF to guide the ablation procedures is an open problem. Electrogram-guided approaches have been proposed. However, electrograms (EGM) are complex and not well-described type of signals and anatomically-based pulmonary vein isolation remains been recommended as the cornerstone procedure. We introduce a method that builds an electroanatomical map to visualize the distribution of different morphological patterns of the EGM signals over the atrial surface. The proposed scheme uses EGM signals recorded with a commercial cardiac mapping. Likewise, two morphological and two non-linear features are computed from each single EGM. Patterns are discriminated using a semi-supervised clustering approach that does not need a priory definition of EGM morphologies or classes. The method was tested under two scenarios: a set of EGM signals recorded in AF patients and a set of signals obtained from 2D simulations of atrial conduction sustained by rotors. Our method was able to locate the clusters in a map of the atrial surface of each patient. These locations allow the specialist to study the distribution of critical AF sites. The method was able to locate the pivot point of the rotors in the 2D models. Our results suggest that the proposed method is a potential assisting tool for guided ablation procedures. Further clinical studies are needed to establish the relationship between clusters and arrhythmogenic substrates in AF, and to validate the usefulness of the method to locate critical conduction sites in patients.


A novel PEGylated liposome-encapsulated SANT75 suppresses tumor growth through inhibiting hedgehog signaling pathway.
The Hedgehog (Hh) pathway inhibitors have shown great promise in cancer therapeutics. SANT75, a novel compound we previously designed to specially inhibit the Smoothened (SMO) protein in the Hh pathway, has greater inhibitory potency than many of commonly used Hh inhibitors. However, preclinical studies of SANT75 revealed water insolubility and acute toxicity. To overcome these limitations, we developed a liposomal formulation of SANT75 and investigated its antitumor efficacy in vitro and in vivo. We encapsulated SANT75 into PEGylated liposome and the mean particle size distribution and zeta-potential (ZP) of liposomes were optimized. Using the Shh-light2 cell and Gli-GFP or Flk-GFP transgenic reporter zebrafish, we confirmed that liposome-encapsulated SANT75 inhibited Hh activity with similar potency as the original SANT75. SANT75 encapsulated into liposome exerted strong tumor growth-inhibiting effects in vitro and in vivo. In addition, the liposomal SANT75 therapy efficiently improved the survival time of tumor-bearing mice without obvious systemic toxicity. The pathological morphology and immunohistochemistry staining revealed that liposomal SANT75 induced tumor cell apoptosis, inhibited tumor angiogenesis as assessed by CD31 and down-regulated the expression of Hh target protein Gli-1 in tumor tissues. Our findings suggest that liposomal formulated SANT75 has improved solubility and bioavailability and should be further developed as a drug candidate for treating tumors with abnormally high Hh activity.


Effects of antibacterials on the unborn child: what is known and how should this influence prescribing.
Antibacterials are among the most commonly prescribed drugs worldwide. In general, infections occur in pregnant women at much the same rate as in the general population. However, as a result of physiological changes brought about by pregnancy, some infections, such as those of the urinary tract, may have an increased incidence. It is important to remember that almost every drug crosses the placenta, ensuring that the unborn fetus is also exposed. When prescribing an antibacterial agent to a pregnant woman, it is important that the mother is treated appropriately while at the same time protecting the unborn child. Certain factors need to be addressed, such as the possible teratogenic risk, changes in pharmacokinetics and the potential toxicity of the drug. In this paper we have reviewed various classes of antibacterials which are commonly used during pregnancy, including penicillins, beta-lactam inhibitors, cephalosporins, macrolides, aminoglycosides, tetracyclines, lincosamides, fluoroquinolones, sulfonamides, nitrofurans, and anti-tubercular agents. Some of these drugs have been on the market for many years, whereas others are relatively new and increasingly popular, despite the fact that the older drugs remain very effective. After reviewing the evidence-based information from epidemiological studies, it appears that most antibacterial agents can be used relatively safely during pregnancy. Women who are pregnant should not be denied appropriate antibacterial therapy because of a lack of information. It is possible to treat the mother, while protecting the unborn child, by prescribing an agent that the causative bacteria is sensitive to, rather than a perceived 'safer' option that may not effectively treat the infection and which may also add to the growing problem of bacterial resistance.


Global, regional, and national burden of tuberculosis, 1990-2016: results from the Global Burden of Diseases, Injuries, and Risk Factors 2016 Study.
Although a preventable and treatable disease, tuberculosis causes more than a million deaths each year. As countries work towards achieving the Sustainable Development Goal (SDG) target to end the tuberculosis epidemic by 2030, robust assessments of the levels and trends of the burden of tuberculosis are crucial to inform policy and programme decision making. We assessed the levels and trends in the fatal and non-fatal burden of tuberculosis by drug resistance and HIV status for 195 countries and territories from 1990 to 2016. We analysed 15 943 site-years of vital registration data, 1710 site-years of verbal autopsy data, 764 site-years of sample-based vital registration data, and 361 site-years of mortality surveillance data to estimate mortality due to tuberculosis using the Cause of Death Ensemble model. We analysed all available data sources, including annual case notifications, prevalence surveys, population-based tuberculin surveys, and estimated tuberculosis cause-specific mortality to generate internally consistent estimates of incidence, prevalence, and mortality using DisMod-MR 2.1, a Bayesian meta-regression tool. We assessed how the burden of tuberculosis differed from the burden predicted by the Socio-demographic Index (SDI), a composite indicator of income per capita, average years of schooling, and total fertility rate. Globally in 2016, among HIV-negative individuals, the number of incident cases of tuberculosis was 9·02 million (95% uncertainty interval [UI] 8·05-10·16) and the number of tuberculosis deaths was 1·21 million (1·16-1·27). Among HIV-positive individuals, the number of incident cases was 1·40 million (1·01-1·89) and the number of tuberculosis deaths was 0·24 million (0·16-0·31). Globally, among HIV-negative individuals the age-standardised incidence of tuberculosis decreased annually at a slower rate (-1·3% [-1·5 to -1·2]) than mortality did (-4·5% [-5·0 to -4·1]) from 2006 to 2016. Among HIV-positive individuals during the same period, the rate of change in annualised age-standardised incidence was -4·0% (-4·5 to -3·7) and mortality was -8·9% (-9·5 to -8·4). Several regions had higher rates of age-standardised incidence and mortality than expected on the basis of their SDI levels in 2016. For drug-susceptible tuberculosis, the highest observed-to-expected ratios were in southern sub-Saharan Africa (13·7 for incidence and 14·9 for mortality), and the lowest ratios were in high-income North America (0·4 for incidence) and Oceania (0·3 for mortality). For multidrug-resistant tuberculosis, eastern Europe had the highest observed-to-expected ratios (67·3 for incidence and 73·0 for mortality), and high-income North America had the lowest ratios (0·4 for incidence and 0·5 for mortality). If current trends in tuberculosis incidence continue, few countries are likely to meet the SDG target to end the tuberculosis epidemic by 2030. Progress needs to be accelerated by improving the quality of and access to tuberculosis diagnosis and care, by developing new tools, scaling up interventions to prevent risk factors for tuberculosis, and integrating control programmes for tuberculosis and HIV. Bill & Melinda Gates Foundation.


THz-Frequency Modulation of the Hubbard U in an Organic Mott Insulator.
We use midinfrared pulses with stable carrier-envelope phase offset to drive molecular vibrations in the charge transfer salt ET-F_{2}TCNQ, a prototypical one-dimensional Mott insulator. We find that the Mott gap, which is probed resonantly with 10 fs laser pulses, oscillates with the pump field. This observation reveals that molecular excitations can coherently perturb the electronic on-site interactions (Hubbard U) by changing the local orbital wave function. The gap oscillates at twice the frequency of the vibrational mode, indicating that the molecular distortions couple quadratically to the local charge density.


Impacts of Watershed Characteristics and Crop Rotations on Winter Cover Crop Nitrate-Nitrogen Uptake Capacity within Agricultural Watersheds in the Chesapeake Bay Region.
The adoption rate of winter cover crops (WCCs) as an effective conservation management practice to help reduce agricultural nutrient loads in the Chesapeake Bay (CB) is increasing. However, the WCC potential for water quality improvement has not been fully realized at the watershed scale. This study was conducted to evaluate the long-term impact of WCCs on hydrology and NO3-N loads in two adjacent watersheds and to identify key management factors that affect the effectiveness of WCCs using the Soil and Water Assessment Tool (SWAT) and statistical methods. Simulation results indicated that WCCs are effective for reducing NO3-N loads and their performance varied based on planting date, species, soil characteristics, and crop rotations. Early-planted WCCs outperformed late-planted WCCs on the reduction of NO3-N loads and early-planted rye (RE) reduced NO3-N loads by ~49.3% compared to the baseline (no WCC). The WCCs were more effective in a watershed dominated by well-drained soils with increased reductions in NO3-N fluxes of ~2.5 kg N·ha-1 delivered to streams and ~10.1 kg N·ha-1 leached into groundwater compared to poorly-drained soils. Well-drained agricultural lands had higher transport of NO3-N in the soil profile and groundwater due to increased N leaching. Poorly-drained agricultural lands had lower NO3-N due to extensive drainage ditches and anaerobic soil conditions promoting denitrification. The performance of WCCs varied by crop rotations (i.e., continuous corn and corn-soybean), with increased N uptake following soybean crops due to the increased soil mineral N availability by mineralization of soybean residue compared to corn residue. The WCCs can reduce N leaching where baseline NO3-N loads are high in well-drained soils and/or when residual and mineralized N availability is high due to the cropping practices. The findings suggested that WCC implementation plans should be established in watersheds according to local edaphic and agronomic characteristics for reducing N leaching.


Processing of cholecystokinin by isolated liver cells.
Although hepatic uptake of cholecystokinin (CCK) has been demonstrated, the liver cell involved and the mechanism of uptake remain unclear. We have used dispersed rat hepatocytes, Kupffer cells, and hepatic endothelial cells to characterize uptake and metabolism of radiolabeled CCK peptides. Only rat hepatocytes showed significant uptake of 125I-labeled cholecystokinin octapeptide (125I-CCK-8). Peptide specificity of uptake by hepatocytes was similar to that seen in the isolated perfused rat liver, with extraction of 125I-CCK-8 being sevenfold greater than that of 125I-CCK-33. Uptake was saturable, as 10(-4) M CCK-4 inhibited uptake of 125I-CCK-8 by 85%. Uptake was rapid, temperature dependent, and extensive and was decreased by metabolic inhibition, a proteolytic enzyme (trypsin), organic anions (sulfobromophthalein and taurocholic acid), and an inhibitor of anion transport 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. In addition, uptake was dependent on extracellular anions but not on extracellular sodium, calcium, or magnesium. After uptake, hepatocytes released radiolabel in a time- and temperature-dependent manner, predominantly in metabolized forms. Thus the hepatocyte is the liver cell that extracts CCK by an active, anion-dependent process. The characteristics of the uptake process resemble those described for organic anions and small, cyclic peptides and suggest that small, linear peptides may undergo hepatocyte extraction by a similar mechanism.


Characterization and assessment of nanoencapsulated sanguinarine chloride as a potential treatment for melanoma.
Sanguinarine has a history of use in both folk medicine and early dermatology for the treatment of cutaneous neoplasms. Applied indiscriminately, bloodroot is an escharotic agent with potential to cause extensive tissue necrosis. However, when used in a controlled fashion, sanguinarine imparts selective cytotoxic/anti-proliferative activity through multiple mechanisms against human/ murine melanoma. To exploit sanguinarine's observed activity against melanoma, a targeted delivery system is required. We present a sol-gel based nanoparticulate platform for encapsulating sanguinarine chloride(sang-np)-a targeted therapeutic capable of steady, reliable delivery of predictable quantities of drug over a sustained time period with minimal undesirable effects. Size and release kinetics of sang-np were characterized using dynamic light scattering and ultraviolet-visible spectroscopy respectively. In vitro efficacy of sang-np was assessed. At both 2 and 24 hours, free sanguinarine killed > 90% of B16 melanoma cells, assessed via MTT assay. At 2 hours, sang-np killed a portion of melanoma cells, increasing to percentages comparable to free sanguinarine by 24 hours. Control(empty) nanoparticles exerted minimal toxicity to melanoma cells at both time points. TUNEL assay revealed that treatment with both sanguinarine and sang-np induces apoptosis in B16 melanoma cells, suggesting that both treatments act via the same mechanism of action. These data confirm controlled release of sanguinarine from sang-np, as well as comparable efficacy and mechanism of action to sanguinarine alone. This suggests that nanoparticle delivery of sanguinarine may be a unique approach to capitalize on this potent agent's inherent anti-tumor activity and overcome many of the limitations with its current formulation.


Interleukin-6-induced Twist and N-cadherin enhance melanoma cell metastasis.
Melanoma patients frequently have elevated serum levels of interleukin-6 (IL-6), which is correlated with a poor prognosis. IL-6 activates STAT3 phosphorylation, inducing the transcription of genes that regulate tumor cell proliferation and antiapoptosis. In addition, recent evidence suggests that IL-6 induces the epithelial-to-mesenchymal transition and enhances the invasiveness of tumor cells of epithelial origin. However, it is unknown whether IL-6 affects mesenchymal tumor cells. In this study, we examined the effects of IL-6 on melanoma cells and found that IL-6 can enhance their metastatic potential by regulating the expression of Twist and N-cadherin. First, we confirmed that human melanoma tissues express IL-6 (especially at the lesion site), the IL-6 receptor, N-cadherin, and nuclear Twist. Next, we found that IL-6 induces STAT3 phosphorylation in WM-266-4 human melanoma cells, resulting in transient upregulation of Twist, which is a key regulator of metastasis. Importantly, the expression of N-cadherin, a protein downstream of Twist, was also increased on the cell surface after treatment with IL-6. These cells showed enhanced invasiveness, assessed using an invasion assay, and formed more metastatic nodules in the lungs of NOD-SCID mice after an intravenous injection. Importantly, melanoma cells with knocked-down N-cadherin formed less lung nodules compared with control in the NOD-SCID mouse model. Our data suggest that increased serum IL-6 in cancer patients could increase the invasiveness of melanoma cells and accelerate metastasis. Blocking IL-6 in the melanoma microenvironment may therefore inhibit disease progression.


EBV-positive diffuse large B-cell lymphoma of the elderly: 2016 update on diagnosis, risk-stratification, and management.
Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) of the elderly is a provisional entity included in the 2008 WHO classification of lymphoid neoplasms. It is a disease typically seen in the elderly and thought to be associated with chronic EBV infection and severe immunosuppression with a component of immunosenescence. Recent research, however, has suggested that EBV-positive DLBCL can be seen in younger, immunocompetent patients. The diagnosis of EBV-positive DLBCL of the elderly is made through a careful pathological evaluation. The differential diagnosis includes infectious mononucleosis (specifically in younger patients), lymphomatoid granulomatosis, Hodgkin lymphoma, and gray zone lymphoma, among others. Detection of EBV-encoded RNA (EBER) is considered standard for diagnosis; however, a clear cutoff for positivity has not been defined. The International Prognostic Index (IPI), and the Oyama score can be used for risk-stratification. The Oyama score includes age >70 years and presence of B symptoms. The expression of CD30 is emerging as a potential adverse, and targetable, prognostic factor. Patients with EBV-positive DLBCL should be staged and managed following similar guidelines than patients with EBV-negative DLBCL. It has been suggested, however, that EBV-positive patients have a worse prognosis than EBV-negative counterparts in the era of chemoimmunotherapy. There is an opportunity to study and develop targeted therapy in the management of patients with EBV-positive DLBCL.


Norms of German adolescents for the Harvard Group Scale of Hypnotic Susceptibility, Form A.
The Harvard Group Scale of Hypnotic Susceptibility, Form A (HGSHS:A) has not been explicitly tested on an adolescent population. In this study, the German version of the HGSHS:A was administered to 99 German adolescents aged 15 to 19. In contrast to other studies, the gender distribution was relatively balanced: 57% female and 43% male. Results were comparable to 14 earlier studies with regard to distribution, mean, and standard deviation. Some peculiarities in contrast to the 14 previous studies are pointed out. It is concluded that the HGSHS:A can be used as a valid and reliable instrument to measure hypnotic suggestibility in adolescent samples.


Use of e-Health services between 1999 and 2002: a growing digital divide.
To evaluate the patterns of e-Health use over a four-year period and the characteristics of users. Longitudinal, population-based study (1999-2002) of members of a prepaid integrated delivery system. Available e-Health services included ordering prescription drug refills, scheduling appointments, and asking medical questions. Rates of known access to e-Health services, and of e-Health use each quarter. The number of members with known e-Health access increased from 51,336 (1.6%) in 1999 to 324,522 (9.3%), in 2002. The percentage of households in which at least one person in the household had access increased from 2.7% to 14.1%. Among the subjects with known access, the percentage of subjects that used e-Health at least once increased from 25.7% in 1999 to 36.2% in 2002. In the multivariate analysis, subjects who had a low expected clinical need, were nonwhite, or lived in low socioeconomic status (SES) neighborhoods were less likely to have used e-Health services in 2002. Disparities by race/ethnicity and SES persisted after controlling for access to e-Health and widened over time. Access to and use of e-Health services are growing rapidly. Use of these services appears to be greatest among persons with more medical need. The majority of subjects, however, do not use any e-Health services. More research is needed to determine potential reasons for disparities in e-Health use by race/ethnicity and SES as well as the implications of these disparities on clinical outcomes.


Surgery in Jehovah's Witnesses - our experience.
Surgeons face a special challenge in treating Jehovah's Witnesses who refuse blood transfusion. To present our surgical experience with this group of patients operated on in our department. A retrospective study of 16 unselected Jehovah's Witnesses patients was conducted between October 2004 and February 2012. We analysed gender, age, haemogram before and after surgery, types of surgery, postoperative complications and the need for blood transfusion, and/or other drugs stimulating erythrogenesis. Eighty-one percent of patients were women; the average age of all patients was 57.3 years. Mean haemoglobin level, preoperative, postoperative, and on the day of discharge from hospital, was 12.5 g/dl, 9.7 g/dl, and 9.29 g/dl, respectively. Over the same time period, mean red blood cell count was 4.53 mln/µl, 3.58 mln/µl, and 3.37 mln/µl, respectively. Two out of 16 patients agreed to have blood transfusion. Drugs used for erythropoiesis stimulation included rEPO, ferrum, and folic acid. No surgical death was noted. We found that abdominal surgery was safe in our small group of Jehovah's Witness patients. However, all Jehovah's Witness patients should be fully informed about the type of procedure and possible consequences of blood transfusion refusal. Two of our patients agreed to blood transfusion in the face of risk of death.


Three synthetic vitamin D analogues induce prostate-specific acid phosphatase and prostate-specific antigen while inhibiting the growth of human prostate cancer cells in a vitamin D receptor-dependent fashion.
Numerous studies have indicated that the secosteroid hormone 1alpha, 25-dihydroxyvitamin D3 protects against the development of clinical prostate cancer (PC). Whether this hormone also has therapeutic potential for patients with advanced PC has not yet been evaluated. Several synthetic vitamin D analogues are now available that have reduced hypercalcemic effects and yet effectively induce differentiation in some cell types. For these reasons, these analogues may be safer and more effective for cancer therapy than the natural hormone. In the current study, 13 such analogues were screened for their abilities to inhibit the growth of PC cell lines. Three of the most consistently effective analogues (Ro 23-7553, Ro 24-5531, and Ro 25-6760) were then chosen for further analysis. Growth studies using clones of the JCA-1 cell line that were transfected with the vitamin D receptor cDNA indicate that the antiproliferative effects of these analogues require vitamin D receptor expression. Furthermore, these three analogues induce the secretion of prostate-specific acid phosphatase and prostate-specific antigen (two markers of the differentiated prostatic phenotype) in the cell line LNCaP. These in vitro studies suggest that Ro 23-7553, Ro 24-5531, and Ro 25-6760 should be further evaluated as therapeutic agents for the treatment of PC.


Biosecurity and the management of emergency animal disease among commercial beef producers in New South Wales and Queensland (Australia).
Australia places great importance on the prevention and management of emergency animal diseases (EAD), with strict quarantine measures offshore and at the border. Livestock producers are crucial for disease control onshore; however, limited information is available on commercial livestock producers' practices in relation to the management of disease risks. The aims of this paper are to investigate how commercial beef producers in Australia's Northern and Southern beef zones manage EADs and to identify drivers for effective biosecurity and EAD prevention. This paper forms part of a broader mixed methods research project involving an analysis of literature and current policies, qualitative semi-structured interviews with government and industry stakeholders and a cross-sectional study among beef producers. The cross-sectional study used a postal survey (n=182) and face-to-face interviews (n=34) to gather data on beef producers' knowledge and practices on biosecurity and EADs and their communication networks. Findings indicate that producers are uncertain about the roles and responsibilities of stakeholders involved in biosecurity and EAD management. This uncertainty may create confusion about EAD management and impact upon producers' willingness to report animal disease, with over 20% reporting the last veterinary contact more than five years ago and an additional 8.5% who had never contacted a veterinarian. Producers had a generally high awareness of the key sources of animal disease risk and they prioritise herd health planning as part of their everyday practices. Over 40% of producers had limited knowledge of the meaning of EAD; and EAD and biosecurity planning was given a low priority, primarily due to the perceived limited likelihood of an EAD event in Australia and the belief that EAD prevention is primarily the role of government. Only a moderate implementation of biosecurity practices, such as isolating incoming animals, having a single property entry point or keeping visitors' records was reported. If faced with an unusual disease event, most producers would contact their private or government veterinarian; however, in some instances most would also treat themselves and over a third would do nothing. Findings from this study suggest that there is a need for better coordination between stakeholders to encourage a shared biosecurity and EAD understanding and to communicate a consistent message to all stakeholders including producers. Further, there is a need for improving producer awareness of the importance of EAD prevention and biosecurity practices as well as the stakeholders' roles within the broader animal health system.


National Centre of Rural Medicine in Norway: a bridge from rural practice to the academy.
Rural medical practice in Norway has an honourable 400 year history, but this has diminished since the end of World War II. Despite official intention to support a decentralised population, rural and remote populations have continuously reduced in Norway over the last 10 years. A consequence of the accompanying reduction in rural and remote GP services has been a distinct reduction in opportunities for medical student and intern placements. In 1999 the University of Tromso implemented some projects to stimulate rural medical practice, funded by the government. This culminated in the 2007 foundation of the Norwegian National Centre of Rural Medicine (NCRM) in Tromso. A key challenge of the NCRM is to identify factors that influence young doctors to choose rural careers. This is reflected in the three concurrent aims or perspectives of the NCRM: (1) to bridge the gap between the academy and rural medical practice (the principal perspective); (2) to promote research, education and networking among rural health professionals (the operational perspective); and (3) to contribute to the recruitment, stability and quality of rural health care (the political perspective). The NCRM has had a number of achievements that include a publication that provides a narrative perspective on rural practice, the role of the rural doctor, and how rural culture and context influence proper clinical decision-making. Another achievement is a professional development and research program that has been successful in fostering a number of major studies, and led to the formation of a supportive PhD research group. The NCRM has also facilitated networking between rural practitioners and academics, at conferences and via its rural doctor website, and promoted cooperative international activities. In these ways the NCRM has fostered the transformation of rural doctors' experience into theory to enhance medical knowledge, begun to redress the balance between community- and hospital-based services, and so made a favourable start to building a bridge between rural practice and the medical academy in Norway.


Partition of circulating lead between serum and red cells is different for internal and external sources of lead.
Serum lead, whole blood lead, and lead in both tibia and calcaneus were measured in each of 49 active lead workers. Serum lead correlated more strongly with both in vivo bone lead measurements than did whole blood lead. The ratio of serum lead to whole blood lead varied from 0.8% to 2.5% and showed a positive correlation with tibia, and an even stronger correlation with calcaneus lead. This implies that lead released from bone (endogenous exposure) results in a higher proportion of whole blood lead being in serum than is the case for exogenous exposure. This observation needs to be confirmed, and the relationships amongst the parameters must be studied further, particularly in former or retired lead workers. If confirmed, since at least a portion of lead in serum is readily diffusible and thus toxicologically more immediately significant than lead bound to red cells, the health implications of endogenous exposure may have to be reassessed.


Two Hsp70 family members expressed in atherosclerotic lesions.
Gene expression profiling was carried out comparing Con A elicited peritoneal macrophages from C57BL6 and FVBN wild-type and apolipoprotein (apo)E knockout mice. An EST, was expressed at higher levels in C57BL6 compared with FVBN mice. mapped to an atherosclerosis susceptibility locus on chromosome 19 revealed in an intercross between atherosclerosis-susceptible C57BL6 and atherosclerosis-resistant FVBN apoE knockout mice. A combination of database search and Northern analysis confirmed that corresponded to 3'-UTR of a hitherto predicted gene, named HspA12A. Blasting the National Center for Biotechnology Information database revealed a closely related homologue, HspA12B. HspA12A and -B have very close human homologues. TaqMan analysis confirmed the increased HspA12A expression (2.6-fold) in elicited peritoneal macrophages from C57BL6 compared with FVBN mice. TaqMan analysis also revealed increased HspA12A and HspA12B expression (87- and 6-fold, respectively) in lesional versus nonlesional portions of the thoracic aorta from C57BL6 apoE knockout mice on a chow diet. In situ hybridization confirmed that both genes were expressed within lesions but not within nonlesional aortic tissue. Blasting of HspA12A and HspA12B against the National Center for Biotechnology Information database (NR) revealed a hit with the Conserved Domain database for Hsp70 (pfam00012.5, Hsp70). Both genes appear to contain an atypical Hsp70 ATPase domain. The BLAST search also revealed that both genes were more similar to primitive eukaryote and prokaryote than mammalian Hsp70s, making these two genes distant members of the mammalian Hsp70 family. In summary, we describe two genes that code for a subfamily of Hsp70 proteins that may be involved in atherosclerosis susceptibility.


Ligation of B7-1/B7-2 by human CD4+ T cells triggers indoleamine 2,3-dioxygenase activity in dendritic cells.
Human monocyte-derived dendritic cells (DCs) are capable of expressing the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO), which allows them to suppress Ag-driven proliferation of T cells in vitro. In DCs that express IDO, the activity of the enzyme is tightly regulated, with the protein being constitutively expressed, but functional activity requiring an additional set of triggering signals supplied during Ag presentation. We now show that triggering of functional IDO obligately requires ligation of B7-1/B7-2 molecules on the DCs by CTLA4/CD28 expressed on T cells. When this interaction was disrupted, IDO remained in the inactive state, and the DCs were unable to inhibit T cell proliferation. Inhibition could be fully restored by direct Ab-mediated cross-linking of B7-1/B7-2. Although both CD4(+) and CD8(+) T cells were susceptible to inhibition once IDO was induced, the ability to trigger functionally active IDO was strictly confined to the CD4(+) subset. Thus, the ability of CD4(+) T cells to induce IDO activity in DCs allowed the CD4(+) population to dominantly inhibit proliferation of the CD8(+) population via the bridge of a conditioned DC. We hypothesize that IDO activation via engagement of B7-1/B7-2 molecules on DCs, specifically, engagement by CTLA4 expressed on regulatory CD4(+) T cells, may function as a physiologic regulator of T cell responses in vivo.


Assessing genetic gain, inbreeding, and bias attributable to different flock genetic means in alternative sheep sire referencing schemes.
Flocks participating in sire referencing schemes can achieve greater genetic gains than those achievable by within-flock selection. However, requirements for joining these schemes can be prohibitive to some producers. The objectives of this study were to determine whether less restrictive schemes or schemes of shorter duration could achieve rates of gain and reduce inbreeding as efficiently as continuous sire referencing schemes (SRS) and to investigate whether bias from different genetic means could be reduced by these alternative schemes. Pedigree and performance data for a single trait with a within-flock heritability of 0.25 were simulated (50 replications) for 15 flocks with 40 to 140 ewes per flock. Founder genetic means for each flock were sampled from a normal distribution with mean 0 and SD equal to the trait's genetic SD. After 10 yr of random mating, flocks had the opportunity to join an SRS and begin selection for the simulated trait. Yearling rams were chosen as reference sires randomly from the top one-sixth of the population ranked on BLUP EBV. Every year, in each flock, 3 reference sires were mated to 10 ewes. Six sire referencing scenarios were considered, in which all flocks participated in a SRS for 1) 15 yr; 2) 5 yr before discontinuing the scheme; 3) 10 yr before discontinuing the scheme; 4) 2 out of every 3 yr; 5) 15 yr with reference sire mating by natural service; and 6) no years (no use of SRS). Ewes not mated to reference sires were mated either to their own home-bred sires exclusively or to a mixture of homebred and unrelated purchased rams of unknown merit. Genetic gain was equivalent whether the SRS used AI or natural service matings, although inbreeding was lower with natural service. Across all scenarios, genetic gain and inbreeding were greater when excess ewes were mated exclusively to homebred sires. Genetic gains without SRS were 80 to 82% lower than when the scheme operated for 15 yr, whereas inbreeding was considerably greater. Other scenarios were intermediate in both gain and inbreeding levels. In all SRS scenarios, bias in EBV attributable to differing flock genetic means rapidly decreased in the first 5 yr of sire referencing. Levels of bias did not substantially increase when flocks discontinued SRS after 5 or 10 yr, suggesting that further participation in an SRS may not be necessary to manage risk. Natural service and noncontinuous SRS are viable options to continuous AI SRS in terms of genetic gain, inbreeding, and bias reduction.


[Risk factors for permanent kidney damage in children with urinary tract infection].
In children, urinary tract infection (UTI) is a very common disease, and can cause permanent kidney damage. To determine risk factors for permanent kidney damage, in children with UTI. In 337 children with UTI (237 female, mean age 4.2 years) a static renal scintigraphy was performed to assess the presence of permanent kidney damage. The history of vesicoureteral reflux and number of episodes of UTI was obtained. One hundred three children had a history of one episode of infection and the rest had recurrent infections. Permanent kidney damage was observed in 161 children (48%). This damage was observed in 39% of children of less than one year of age, in 43% of children aged 1 to 5 years of age and in 58% of children older than 5 years (p = 0.02). Sixty three percent of 122 children with vesicoureteral reflux had permanent kidney damage, compared with 27% of children without this condition (p < 0.001). Likewise, damage was observed in 36% of children with one episode of infection and 47% of children with recurrent infections (p < 0.01). No gender differences were observed. Vesicoureteral reflux, recurrence of UTI and age are associated with permanent renal damage in children with UTI.