Development of immunoassays for determination of circulating venom antigens during envenomations by coral snakes (Micrurus species).
A reverse agglutination assay and two capture enzyme-linked immunoassays (ELISAs) for the quantitative determination of Micrurus nigrocinctus nigrocinctus venom antigens in fluids were developed using affinity-purified polyclonal antibodies and a cocktail of three monoclonal antibodies. The lower detection limit was 0.3 mg/ml for the reverse agglutination assay and 4 ng/ml for the capture ELISAs. The optical densities of both ELISAs correlated very well with venom concentrations in the range 4-333 ng/ml (r = 0.99). The ability of these assays to detect venoms of several medically important Micrurus species was studied. Besides detecting homologous venom, both ELISAs were also useful to quantitate venom from M. fulvius, M. dumerilii carinicauda and M. alleni. Using biotinylated polyclonal antibodies, M. n. nigrocinctus venom antigens were detected in sera or plasma from rabbits and mice during experimental envenomations with lethal and sublethal venom doses. The assays described in this work are promising tests to estimate the severity of poisoning in envenomations by the most important coral snakes of North and Central America.


Efficient Removal of Ultrafine Particles from Diesel Exhaust by Selected Tree Species: Implications for Roadside Planting for Improving the Quality of Urban Air.
Human exposure to airborne ultrafine (≪1 μm) particulate pollution may pose substantial hazards to human health, particularly in urban roadside environments where very large numbers of people are frequently exposed to vehicle-derived ultrafine particles (UFPs). For mitigation purposes, it is timely and important to quantify the deposition of traffic-derived UFPs onto leaves of selected plant species, with particularly efficient particle capture (high deposition velocity), which can be installed curbside, proximal to the emitting vehicular sources. Here, we quantify the size-resolved capture efficiency of UFPs from diesel vehicle exhaust by nine temperate-zone plant species, in wind tunnel experiments. The results show that silver birch (79% UFP removal), yew (71%), and elder (70.5%) have very high capabilities for capture of airborne UFPs. Metal concentrations and metal enrichment ratios in leaf leachates were also highest for the postexposure silver birch leaves; scanning electron microscopy showed that UFPs were concentrated along the hairs of these leaves. For all but two species, magnetic measurements demonstrated substantial increases in the concentration of magnetic particles deposited on the leaves after exposure to the exhaust particulates. Together, these new data show that leaf-deposition of UFPs is chiefly responsible for the substantial reductions in particle numbers measured downwind of the vegetation. It is critical to recognize that the deposition velocity of airborne particulate matter (PM) to leaves is species-specific and often substantially higher (∼10 to 50 times higher) than the "standard" Vd values (e.g., 0.1-0.64 cm s-1 for PM2.5) used in most modeling studies. The use of such low Vd values in models results in a major under-estimation of PM removal by roadside vegetation and thus misrepresents the efficacy of selected vegetation species in the substantial (≫20%) removal of PM. Given the potential hazard to health posed by UFPs and the removal efficiencies shown here (and by previous roadside measurements), roadside planting (maintained at or below head height) of selected species at PM "hotspots" can contribute substantially and quickly to improve in urban air quality and reductions in human exposure. These findings can contribute to the development and implementation of mitigation policies of traffic-derived PM on an international scale.


Underoil Superhydrophilic Metal Felt Fabricated by Modifying Ultrathin Fumed Silica Coatings for the Separation of Water-in-Oil Emulsions.
Although various superhydrophobic/superoleophilic porous materials have been developed and successfully applied to separate water-in-oil emulsions through the size-sieving mechanism, the separation performance is restricted by their nanoscale pore size severely. In this study, the wettability of underoil water on fumed silica was experimentally observed, and the underlying mechanism was investigated by carrying out theoretical analysis and molecular dynamic (MD) simulations. Further, we present a novel, facile, and an inexpensive technique to fabricate an underoil superhydrophilic metal felt with microscale pores for the separation of water-in-oil emulsions using SiO2 nanoparticles (NPs) as building blocks. The as-prepared underoil superhydrophilic coating is closed-packed and ultrathin (the thickness is approximately hundreds of nanometers), as well as capable of being coated on a metal felt with complex structures without blocking its pores. The as-prepared metal felt could adsorb water droplets directly from oil, which endowed it with the ability to separate both surfactant-free and surfactant-stabilized water-in-oil emulsions with high separation efficiency up to 99.7% even though its pore size is larger than that of the emulsified droplet. The filtration flux for the separation of span 80-stabilized emulsion is up to ∼4000 L·m-2·h-1. Its separation performance is better than most of the other traditional membranes and superwettable materials used for the separation of water-in-oil emulsions. Moreover, the as-prepared metal felt retained outstanding separation performance even after 30 cycles of use, which demonstrated its excellent reusability and durability. Additionally, the distinctive wettability of underoil superhydrophilicity endued coated metal felt with superior antifouling properties toward crude oil. Overall, this study not only provides a new perspective on separating water-in-oil emulsions but also gives a universal approach to develop unique wettability surfaces.


Effect of circumcision on urinary tract infection after successful antireflux surgery.
To evaluate whether circumcision during antireflux surgery can reduce the incidence of urinary tract infection (UTI) after successful ureteric reimplantation in patients with primary vesico-ureteric reflux (VUR). Children who had undergone antireflux surgery for primary VUR were divided into group 1 (27, circumcised at the time of antireflux surgery at the parents' request) and group 2 (50, those not circumcised). All antireflux operations were by the Cohen method. Regular urine samples were cultured to detect UTI, which was defined as a single species with >10(5) colony-forming units/mL in a midstream voided specimen. Numbers of UTI episodes before and after surgery were compared between the groups, with (99m)Tc-dimercaptosuccinic acid (DMSA) renal scans also taken in all patients. Each scan was blindly reviewed in terms of the size, number and zonal location of cortical defects, based on morphology. Interval changes were categorised as improved, no change, progressed, and new scar formation, and compared between the groups. Prophylactic antibiotics were maintained until the follow-up studies at 4-6 months after surgery. There was no significant difference between the groups in age at the time of operation (mean 42.4 vs 47.4 months), the age at the first documented UTI (mean 26.5 vs 29.3 months), reflux grade, or number of UTI episodes and renal parenchymal scarring on DMSA before surgery. There was no significant difference between the groups in the number of UTI episodes at a mean (range) follow-up of 151.3 (114-207) months after antireflux surgery. Also there was no significant morphological change on follow-up renal scans and no difference between the groups. These findings suggest that circumcision during antireflux surgery has no effect on the incidence of postoperative UTI.


[The comparative analysis of subcellular fractionation methods for revealing of alpha-actinin 1 and alpha-actinin 4 in A431 cells].
Alpha-actinin 1 and alpha-actinin 4 belong to a family of actin-binding proteins with shared structural function and regulation of several processes in a cell. Based on previous data on different distribution of these proteins in the nucleus and cytoplasm, we have explored in detail the distribution of alpha-actinin 1 and alpha-actinin 4 in subcellular fractions in A431 cells spread on fibronectin. Several methods of subcellular fractionation were used. Complex approach allowed resuming that revealing of alpha-actinin isoforms in fractions depended on the composition of lysis buffer and preliminary low-temperature freezing of the cells. We have drawn a conclusion that alpha-actinin 4 can be found in all cytoplasmic and nuclear subfractions, while alpha-actinin 1 is characterized by cytoplasmic and membrane localization with specificity of its distribution tightly to the nuclear membrane.


Classification of genera of Pasteurellaceae using conserved predicted protein sequences.
The aim of the investigation was to investigate the phylogeny of the 49 type strains of species of Pasteurellaceae and three genomospecies, which are available with whole genomic sequences. The genomes were downloaded from National Center for Biotechnological Information and for three species of Avibacterium sequenced in the present investigation. From the predicted protein sequences of proteins, which were conserved in all genomes, 31 proteins were randomly selected for the study. The protein sequences were concatenated for each taxon, and a multiple alignment reconstructed for the 52 taxa. Phylogenetic analysis was performed by using the maximum-likelihood and neighbour-joining methods and confirmed the classification of the genera, which have been classified based on phylogenetic analysis of 16S rRNA gene sequences. The comparison linked [Haemophilus]parainfluezae and [Haemophilus] pittmania with Haemophilus influenzae (type species of genus) although at a much lower level than observed for Haemophilus aegyptius, H. influenzae and Haemophilus haemolyticus. The comparison documented that three, three and nine species of Actinobacillus, Pasteurella and Haemophilus, respectively, are not properly classified at genus level. Similar conclusions have been drawn by 16S rRNA gene sequence comparisons. The highest inter genus pairwise similarity was 88 % based on the comparison of the 31 concatenated protein sequences of the species included in the comparison. The level of intra genus pairwise similarity was also 88 %.


Can this patient read and understand written health information?
Patients with limited literacy are at higher risk for poor health outcomes; however, physicians' perceptions are inaccurate for identifying these patients. To systematically review the accuracy of brief instruments for identifying patients with limited literacy. Search of the English-language literature from 1969 through February 2010 using PubMed, Psychinfo, and bibliographies of selected manuscripts for articles on health literacy, numeracy, reading ability, and reading skill. Prospective studies including adult patients 18 years or older that evaluated a brief instrument for identifying limited literacy in a health care setting compared with an accepted literacy reference standard. Studies were evaluated independently by 2 reviewers who each abstracted information and assigned an overall quality rating. Disagreements were adjudicated by a third reviewer. Ten studies using 6 different instruments met inclusion criteria. Among multi-item measures, the Newest Vital Sign (English) performed moderately well for identifying limited literacy based on 3 studies. Among the single-item questions, asking about a patient's use of a surrogate reader, confidence filling out medical forms, and self-rated reading ability performed moderately well in identifying patients with inadequate or marginal literacy. Asking a patient, "How confident are you in filling out medical forms by yourself?" is associated with a summary likelihood ratio (LR) for limited literacy of 5.0 (95% confidence interval [CI], 3.8-6.4) for an answer of "a little confident" or "not at all confident"; a summary LR of 2.2 (95% CI, 1.5-3.3) for "somewhat confident"; and a summary LR of 0.44 (95% CI, 0.24-0.82) for "quite a bit" or "extremely confident." Several single-item questions, including use of a surrogate reader and confidence with medical forms, were moderately effective for quickly identifying patients with limited literacy.


Incorporating Geriatric Assessment into a Nephrology Clinic: Preliminary Data from Two Models of Care.
Older adults with advanced chronic kidney disease (CKD) experience functional impairment that can complicate CKD management. Failure to recognize functional impairment may put these individuals at risk of further functional decline, nursing home placement, and missed opportunities for timely goals-of-care conversations. Routine geriatric assessment could be a useful tool for identifying older adults with CKD who are at risk of functional decline and provide contextual information to guide clinical decision-making. Two innovative programs were implemented in the Veterans Health Administration that incorporate geriatric assessment into a nephrology visit. In one program, a geriatrician embedded in a nephrology clinic used standardized geriatric assessment tools with individuals with CKD aged 70 and older (Comprehensive Geriatric Assessment for CKD) (CGA-4-CKD). In the second program, a nephrology clinic used comprehensive appointments for individuals aged 75 and older to conduct geriatric assessments and CKD care (Renal Silver). Data on 68 veterans who had geriatric assessments through these programs between November 2013 and May 2015 are reported. In CGA-4-CKD, difficulty with one or more activities of daily living (ADLs), history of falls, and cognitive impairment were each found in 27.3% of participants. ADL difficulty was found in 65.7%, falls in 28.6%, and cognitive impairment in 51.6% of participants in Renal Silver. Geriatric assessment guided care processes in 45.4% (n = 15) of veterans in the CGA-4-CKD program and 37.1% (n = 13) of those in Renal Silver. Findings suggest there is a significant burden of functional impairment in older adults with CKD. Knowledge of this impairment is applicable to CKD management.


Assessment for water quality by artificial neural network in Daya Bay, South China Sea.
In this study, artificial neural network such as a self-organizing map (SOM) was used to assess for the effects caused by climate change and human activities on the water quality in Daya Bay, South China Sea. SOM has identified the anthropogenic effects and seasonal characters of water quality. SOM grouped the four seasons as four groups (winter, spring, summer and autumn). The Southeast Asian monsoons, northeasterly from October to the next April and southwesterly from May to September have also an important influence on the water quality in Daya Bay. Spatial pattern is mainly related to anthropogenic activities and hydrodynamics conditions. In spatial characteristics, the water quality in Daya Bay was divided into two groups by chemometrics. The monitoring stations (S3, S8, S10 and S11) were in these area (Dapeng Ao, Aotou Harbor) and northeast parts of Daya Bay, which are areas of human activity. The thermal pollution has been observed near water body in Daya Bay Nuclear Power Plant (S5). The rest of the monitoring sites were in the south, central and eastern parts of Daya Bay, which are areas that experience water exchanges from South China Sea. The results of this study may provide information on the spatial and temporal patterns in Daya Bay. Further research will be carry out more research concerning functional changes in the bay ecology with respect to changes in climatic factor, human activities and bay morphology in Daya Bay.


Invasion precedes tumor mass formation in a malignant brain tumor model of genetically modified neural stem cells.
Invasiveness, cellular atypia, and proliferation are hallmarks of malignant gliomas. To effectively target each of these characteristics, it is important to understand their sequence during tumorigenesis. However, because most gliomas are diagnosed at an advanced stage, the chronology of gliomagenesis milestones is not well understood. The aim of the present study was to determine the onset of these characteristics during tumor development. Brain tumor-initiating cells (BTICs) were established by overexpressing H-Ras(V12) in normal neural stem/progenitor cells isolated from the subventricular zone of adult mice harboring a homozygous deletion of the Ink4a/Arf locus. High-grade malignant brain tumors were then created by orthotopic implantation of 10(5) BTICs into the forebrain of 6-week-old wild-type mice. Micewere killed every week for 5 weeks, and tumors were assessed for cellular atypia, proliferation, hemorrhage, necrosis, and invasion. All mice developed highly invasive, hypervascular glioblastoma-like tumors. A 100% penetrance rate and a 4-week median survival were achieved. Tumor cell migration along fiber tracts started within days after implantation and was followed by perivascular infiltration of tumor cells with marked recruitment of reactive host cells. Next, cellular atypia became prominent. Finally, mass proliferation and necrosis were observed in the last stage of the disease. Video monitoring of BTICs in live brain slices confirmed the early onset of migration, as well as the main cell migration patterns. Our results showed that perivascular and intraparenchymal tumor cell migration precede tumor mass formation in the adult brain, suggesting the need for an early and sustained anti-invasion therapy.


Contribution of Leydig and Sertoli cells to testosterone production in mouse fetal testes.
Testosterone is a final product of androgenic hormone biosynthesis, and Leydig cells are known to be the primary source of androgens. In the mammalian testis, two distinct populations of Leydig cells, the fetal and the adult Leydig cells, develop sequentially, and these two cell types differ both morphologically and functionally. It is well known that the adult Leydig cells maintain male reproductive function by producing testosterone. However, it has been controversial whether fetal Leydig cells can produce testosterone, and the synthetic pathway of testosterone in the fetal testis is not fully understood. In the present study, we generated transgenic mice in which enhanced green fluorescence protein was expressed under the control of a fetal Leydig cell-specific enhancer of the Ad4BP/SF-1 (Nr5a1) gene. The transgene construct was prepared by mutating the LIM homeodomain transcription factor (LHX9)-binding sequence in the promoter, which abolished promoter activity in the undifferentiated testicular cells. These transgenic mice were used to collect highly pure fetal Leydig cells. Gene expression and steroidogenic enzyme activities in the fetal Leydig cells as well as in the fetal Sertoli cells and adult Leydig cells were analyzed. Our results revealed that the fetal Leydig cells synthesize only androstenedione because they lack expression of Hsd17b3, and fetal Sertoli cells convert androstenedione to testosterone, whereas adult Leydig cells synthesize testosterone by themselves. The current study demonstrated that both Leydig and Sertoli cells are required for testosterone synthesis in the mouse fetal testis.


[Social actors in HIV/AIDS prevention: opposition and interests in educational policy in Mexico, 1994-2000].
Studies and recommendations by health agencies have emphasized the importance of education in HIV-AIDS prevention. Mexico has included topics on sexuality and HIV-AIDS in school programs, triggering resistance by some social actors. The current study seeks to clarify the various positions and interests and their influence on the textbook content. A literature search was conducted on the period during which the last educational reform was implemented in Mexico. The discourse analysis focused on the ethnography of communication, which identified: the various actors' positions, arguments, actions, economic and political power, and relations to others. The results show that those who oppose the inclusion of these themes in the school curriculum base their position on tradition, contrary to modernization and secularization of social life, and that their positions range from refusal to raising conditions. Networks have been formed that provide such groups with significant economic and political power. Government has given in to some demands by partially modifying the textbook contents. The current analysis proposes to reflect on the potential repercussions of such actions on the control of the epidemic.


The Hydrophobic Patch Directs Cyclin B to Centrosomes to Promote Global CDK Phosphorylation at Mitosis.
The cyclin-dependent kinases (CDKs) are the major cell-cycle regulators that phosphorylate hundreds of substrates, controlling the onset of S phase and M phase [1-3]. However, the patterns of substrate phosphorylation increase are not uniform, as different substrates become phosphorylated at different times as cells proceed through the cell cycle [4, 5]. In fission yeast, the correct ordering of CDK substrate phosphorylation can be established by the activity of a single mitotic cyclin-CDK complex [6, 7]. Here, we investigate the substrate-docking region, the hydrophobic patch, on the fission yeast mitotic cyclin Cdc13 as a potential mechanism to correctly order CDK substrate phosphorylation. We show that the hydrophobic patch targets Cdc13 to the yeast centrosome equivalent, the spindle pole body (SPB), and disruption of this motif prevents both centrosomal localization of Cdc13 and the onset of mitosis but does not prevent S phase. CDK phosphorylation in mitosis is compromised for approximately half of all mitotic CDK substrates, with substrates affected generally being those that require the highest levels of CDK activity to become phosphorylated and those that are located at the SPB. Our experiments suggest that the hydrophobic patch of mitotic cyclins contributes to CDK substrate selection by directing the localization of Cdc13-CDK to centrosomes and that this localization of CDK contributes to the CDK substrate phosphorylation necessary to ensure proper entry into mitosis. Finally, we show that mutation of the hydrophobic patch prevents cyclin B1 localization to centrosomes in human cells, suggesting that this mechanism of cyclin-CDK spatial regulation may be conserved across eukaryotes.


Changes in the effect of testosterone on hypothalamic nitric oxide synthetase during sexual maturation. Its relationship with GnRH release.
To determine the effect of testosterone administration to prepubertal (15 days old) and peripubertal rats (30 days old) on hypothalamic nitric oxide synthetase (NOS), and GnRH release. Hypothalamic samples containing the anterior preoptic and medial basal areas (APO-MBH) were incubated for 30 minutes in 500 l of Earle's medium with glucose (1 mg/ml) and bacitracin (20 mM). GnRH was determined by RIA in the medium and NOS activity was determined in APO-MBH after 10 min of incubation by the conversion of (14) C arginine to (14) C citrulline. Treatment with testosterone propionate, significantly decreased NOS hypothalamic activity in prepubertal male rats. ( 58.41 +/- 0.85; Testosterone: 25.61 +/- 1.40, p<0.001) and had no effect in peripubertal male rats (CONTROL 49.28 +/- 1.50; Testosterone 51.48 +/- 5.2 pmoles NO/10 min/hypothalamus). On the other hand, in prepubertal rats the treatment decreased Gn-RH release ( 3.62 +/- 0.23; Testosterone: 1.38 +/- 0.11 (pg/ml medium, p<0.001) and had no effect on Gn-RH release in 30 days old rats ( 3.65 +/- 0.33;Testosterone: 4.15 +/- 0.36 pg/ ml, medium). These results clearly demonstrated that testosterone has an inhibitory effect on hypothalamic NOS activity in prepubertal rats while it did not affect the concentration of this neurotransmitter system in peripubertal rats. This pattern is similar to that observed with GnRH hypothalamic release since testosterone has an inhibitory effect in prepubertal rats and did not modify the GnRH release in peripubertal rats. Taking into account the well known stimulatory effect of NO on GnRH and the decrease in the sensitivity of GnRH-gonadotrophin axis to the inhibitory feedback effect of testosterone during sexual maturation and the onset of puberty, it is proposed that the changes here described are connected with maturational modifications in the sexual hormones on-GnRH axis connected with the onset of puberty.


First Report of Watermelon chlorotic stunt virus in Cucurbits in Lebanon.
In August 2009 in the Marjyoun region in South Lebanon, severe yellowing symptoms on melon (Cucumis melo) and pronounced dwarfing and mosaics on watermelon (Citrullus lanatus) led to significant yield losses. Watermelon chlorotic stunt virus (WmCSV), genus Begomovirus, family Geminiviridae, was suspected. Symptomatic samples were collected close to the end of the growing season from several fields. The small scale CTAB protocol was followed for nucleic acid extraction. Samples were tested by PCR for WmCSV and Squash leaf curl virus (SLCV) using specific primers for SLCV (2) and newly designed WmCSV specific primers: (WMAR1: 5' TTTTCCGACACGATGAGTGAT 3'; WMAF3: 5' ACTGGACTTAGCGCTTTGTAT 3'; amplicon size 1,061 bp). Of 39 symptomatic samples, 90% were infected with WmCSV, 13/14 (93%) melon samples and 22/25 (88%) watermelon samples, while 64% were infected with SLCV, indicating a high incidence of mixed infections. In November 2009, no cucurbits were found in Marjyoun since farmers refrained from planting late crops after devastating losses in the previous year. Therefore, 92 samples were collected from other southern regions and 114 samples from northern regions. All squash samples had leaf curl symptoms, while 75 to 85% of cucumber and melon had yellowing symptoms. No WmCSV was detected in North Lebanon, even though 100% of squash samples and 79% of other cucurbit samples were positive for SLCV. However, in South Lebanon, WmCSV was detected 9/20 (45%) in melon, 12/32 (38%) in cucumber (Cucumis sativus), and 6/40 (15%) in squash; while the incidence of SLCV was high particularly on squash (39/40, 98%) and cucumber (30/32, 94%) followed by melon (7/20, 35%). The survey was repeated in 2010, and the previous year's results were confirmed: no WmCSV was detected in North Lebanon, while 39/40 (98%) melon samples tested in November were positive for SLCV. In southern Lebanon, WmCSV was not detected in melon or watermelon samples collected in June; however, in November it was detected in 11/23 (48%) squash and 9/33 (27%) melon. WmCSV genome was amplified by rolling circle amplification (RCA) using the TempliPhi Amplification Kit (GE Healthcare) The RCA product was sequenced using mostly locally designed primers, and the sequences were submitted to GenBank: WmCSV DNA A: HM368371.1; WmCSV DNA B: HM368372. Phylogenetic analysis showed that WmCSV DNA A was most closely related to isolates from Israel (EF201809.1) and Jordan (EU561237.1), sharing 99% nt identities with both isolates; WmCSV DNA B was found to be most closely related to an isolate from Israel (EF201810.1), with 98% nt identity. WmCSV was first detected in Yemen (4) but was detected quite recently in Israel and Jordan (1). Within a short period, Lebanon experienced the introduction of two new whitefly transmitted begomoviruses. WmCSV seems so far to be restricted only to South Lebanon, while SLCV is widespread. The synergistic interaction between a mixed infection by SLCV and WmCSV in melon resulted in significant symptom enhancement, plant shortening, and up to 54% yield reduction in summer (3). Hence, the development of resistant varieties coupled with the implementation of adapted integrated pest management strategies would be essential for successful production of cucurbit crops. References: (1) Al-Musa et al. J. Phytopathol. 156:311, 2008. (2) Sobh et al. Plant Dis. 26:1231, 2012. (3) Sufrin-Ringwald and Lapidot. Phytopathology 101:281, 2011. (4) Walkey et al. Tropical Pest Management 36:195, 1990.


Quantitative Evaluation of Adult Subglottic Stenosis Using Intraoperative Long-range Optical Coherence Tomography.
To determine the feasibility of long-range optical coherence tomography (LR-OCT) as a tool to intraoperatively image and measure the subglottis and trachea during suspension microlaryngoscopy before and after endoscopic treatment of subglottic stenosis (SGS). Long-range optical coherence tomography of the adult subglottis and trachea was performed during suspension microlaryngoscopy before and after endoscopic treatment for SGS. The anteroposterior and transverse diameters, cross-sectional area (CSA), distance from the vocal cords, and length of the SGS were measured using a MATLAB software. Pre-intervention and postintervention airway dimensions were compared. Three-dimensional volumetric airway reconstructions were generated using medical image processing software (MIMICS). Intraoperative LR-OCT imaging was performed in 3 patients undergoing endoscopic management of SGS. Statistically significant differences in mean anteroposterior diameter (P < .01), transverse diameter (P < .001), and CSA (P < .001) were noted between pre-intervention and postintervention data. Three-dimensional airway models were viewed in cross-sectional format and via virtual "fly through" bronchoscopy. This is the first report of intraoperative LR-OCT of the subglottic and tracheal airway before and after surgical management of SGS in humans. Long-range optical coherence tomography offers a practical means to measure the dimensions of SGS and acquire objective data on the response to endoscopic treatment of SGS.


Development of hematopoietic cancers after implantation of total joint replacement.
The evidence from the 4 epidemiologic studies published before mid 1995, which have investigated the incidence of cancers in patients who have undergone joint implants, is conflicting. The results of the 2 earlier studies suggested a sustained increase in the risk of lymphoma and leukemia after total hip arthroplasty. The results of the 2 more recent studies have not confirmed this, although in 1 study an increased risk was observed in the first year after implantation. The heterogeneity may be statistical in origin, but could also have a biologic explanation in the greater proportion of metal on metal prostheses used before 1973. All 4 studies used national data as the comparison. Here, are presented the results of 2 matched cohort studies and a case control study set in North America and Scotland, and an overview of the 4 previous studies. Neither the results of the matched studies of patients operated on after 1973 nor the results of the latter 2 published studies suggest an increased risk of lymphoma or leukemia. If metal on metal articulations are reintroduced, careful surveillance is essential.


Electronic transfer of prescription-related information: comparing views of patients, general practitioners, and pharmacists.
The National Health Service (NHS) intends to introduce a system of electronic transfer of prescription-related information between general practitioners (GPs) and community pharmacies. The NHS Plan describes how this will be achieved. To gather opinions of patients, GPs, and community pharmacists on the development of a system of electronic transfer of prescription-related information between GPs and community pharmacies. Survey combining interviews, focus groups, and postal questionnaires. General practitioners, opinion leaders, computing experts, pharmacists, and patients. Eight hundred members of the public, 200 GPs, and 200 community pharmacists, all living in Scotland. Content-setting interviews and focus groups were conducted with purposive samples of relevant groups. Postal questionnaires were developed and sent to random samples of members of the public selected from the electoral roll, GPs, and community pharmacists. The corrected postal response rates were: 69% (patients); 74% (GPs); and 74% (community pharmacists). All three groups were generally supportive of electronic transfer of prescription-related information. Different aspects appealed to each group: patients anticipated improved convenience; GPs, better repeat prescribing; and pharmacists, an enhanced professional role. Security of patient-identifiable information was the main concern. All groups acknowledged potential benefits of a full primary care information system, but GPs and patients had reservations about allowing community pharmacists to access parts of the medical record that did not concern medication. Electronic transfer of prescription-related information is likely to be acceptable to all users, but concerns about patient confidentiality and an extended role for pharmacists in prescription management need to be addressed.


Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts.
Two-dimensional carbon-based nanomaterials, including graphene oxide and graphene, are potential candidates for biomedical applications such as sensors, cell labeling, bacterial inhibition, and drug delivery. Herein, we explore the biocompatibility of graphene-related materials with controlled physical and chemical properties. The size and extent of exfoliation of graphene oxide sheets was varied by sonication intensity and time. Graphene sheets were obtained from graphene oxide by a simple (hydrazine-free) hydrothermal route. The particle size, morphology, exfoliation extent, oxygen content, and surface charge of graphene oxide and graphene were characterized by wide-angle powder X-ray diffraction, atomic force microscopy, X-ray photoelectron spectroscopy, dynamic light scattering, and zeta-potential. One method of toxicity assessment was based on measurement of the efflux of hemoglobin from suspended red blood cells. At the smallest size, graphene oxide showed the greatest hemolytic activity, whereas aggregated graphene sheets exhibited the lowest hemolytic activity. Coating graphene oxide with chitosan nearly eliminated hemolytic activity. Together, these results demonstrate that particle size, particulate state, and oxygen content/surface charge of graphene have a strong impact on biological/toxicological responses to red blood cells. In addition, the cytotoxicity of graphene oxide and graphene sheets was investigated by measuring mitochondrial activity in adherent human skin fibroblasts using two assays. The methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, a typical nanotoxicity assay, fails to predict the toxicity of graphene oxide and graphene toxicity because of the spontaneous reduction of MTT by graphene and graphene oxide, resulting in a false positive signal. However, appropriate alternate assessments, using the water-soluble tetrazolium salt (WST-8), trypan blue exclusion, and reactive oxygen species assay reveal that the compacted graphene sheets are more damaging to mammalian fibroblasts than the less densely packed graphene oxide. Clearly, the toxicity of graphene and graphene oxide depends on the exposure environment (i.e., whether or not aggregation occurs) and mode of interaction with cells (i.e., suspension versus adherent cell types).


Neurotransmitter synthesis in poststroke cortical neurogenesis in adult rats.
Neurogenesis occurs in the cerebral cortex of adult rats after focal cerebral ischemia. Whether or not the newborn neurons could synthesize neurotransmitters is unknown. To elucidate such a possibility, a photothrombotic ring stroke model with spontaneous reperfusion was induced in adult male Wistar rats. The DNA duplication marker BrdU was repeatedly injected, and the rats were sacrificed at various times after stroke. To detect BrdU nuclear incorporation and various neurotransmitters, brain sections were processed for single/double immunocytochemistry and single/double/triple immunofluorescence. Stereological cell counting was performed to assess the final cell populations. At 48 h, 5 days, 7 days, 30 days, 60 days and 90 days after stroke, numerous cells were BrdU-immunolabeled in the penumbral cortex. Some of these were doubly immunopositive to the cholinergic neuron-specific marker ChAT or GABAergic neuron-specific marker GAD. As analyzed by 3-D confocal microscopy, the neurotransmitters acetylcholine and GABA were colocalized with BrdU in the same cortical cells. In addition, GABA was colocalized with the neuron-specific marker Neu N in the BrdU triple-immunolabeled cortical cells. This study suggests that the newborn neurons are capable of synthesizing the neurotransmitters acetylcholine and GABA in the penumbral cortex, which is one of the fundamental requisites for these neurons to function in the poststroke recovery.


Development and validation of Skindex-Teen, a quality-of-life instrument for adolescents with skin disease.
To develop and validate a quality-of-life index for adolescents with skin disease. Prospective, longitudinal cohort study taking place from July 15, 2005, through February 29, 2008. Academic pediatric dermatology practice. A total of 205 patients between the ages of 12 and 17 years with a skin condition. Participants completed the Skindex-Teen questionnaire, reported sociodemographic data, and answered clinical questions at enrollment. Four weeks later, they completed Skindex-Teen and responded to clinical questions again. Forty patients also completed Skindex-Teen 48 hours after baseline. Of 205 patients, 200 (97.6%) completed all enrollment materials; 125 (62.5%) of these returned follow-up materials, of which 115 (57.5%) were complete. Twenty-three of 40 participants (57.5%) completed the 48-hour-after-initial-answers materials. The mean (SD) age of the patients was 15.3 (1.4) years (range, 12.9-18.0 years). Approximately half reported their skin condition as poor (26 [13.0%]) or fair (67 [33.5%]); most rated their general health as very good (62 [31.0%]) or excellent (80 [40.0%]). The dimensionality of the items was evaluated using factor analysis; results suggested 2 factors, termed physical symptoms (5 items) and psychosocial functioning (16 items). Both scales demonstrated excellent internal consistency. All item-scale reliabilities were greater than 0.4. Fit of items to the measurement model was supported by Rasch analysis. Test-retest reliability was supported by intraclass correlation coefficients (0.82-0.94) for total scores, physical symptoms, and psychosocial functioning scales. At 4 weeks, 16 (13.9%) reported worsening of their skin condition, 57 (19.6%) reported it had remained unchanged, and 42 (36.5%) reported improvement. Significant mean differences were present between the improved and worsened groups for the psychosocial functioning scale and total scores. These data provide evidence of construct, content, and face validity for Skindex-Teen, a new quality-of-life instrument for adolescents with skin disease. Test-retest reliability was verified, and responsiveness at 4 weeks was shown for total scores and the psychosocial functioning scale scores.


The gut microbiota metabolism of pomegranate or walnut ellagitannins yields two urolithin-metabotypes that correlate with cardiometabolic risk biomarkers: Comparison between normoweight, overweight-obesity and metabolic syndrome.
Urolithins are microbial metabolites produced after consumption of ellagitannin-containing foods such as pomegranates and walnuts. Parallel to isoflavone-metabolizing phenotypes, ellagitannin-metabolizing phenotypes (urolithin metabotypes A, B and 0; UM-A, UM-B and UM-0, respectively) can vary among individuals depending on their body mass index (BMI), but correlations between urolithin metabotypes (UMs) and cardiometabolic risk (CMR) factors are unexplored. We investigated the association between UMs and CMR factors in individuals with different BMI and health status. UM was identified using UPLC-ESI-qToF-MS in individuals consuming pomegranate or nuts. The associations between basal CMR factors and the urine urolithin metabolomic signature were explored in 20 healthy normoweight individuals consuming walnuts (30 g/d), 49 healthy overweight-obese individuals ingesting pomegranate extract (450 mg/d) and 25 metabolic syndrome (MetS) patients consuming nuts (15 g-walnuts, 7.5 g-hazelnuts and 7.5 g-almonds/d). Correlations between CMR factors and urolithins were found in overweight-obese individuals. Urolithin-A (mostly present in UM-A) was positively correlated with apolipoprotein A-I (P ≤ 0.05) and intermediate-HDL-cholesterol (P ≤ 0.05) while urolithin-B and isourolithin-A (characteristic from UM-B) were positively correlated with total-cholesterol, LDL-cholesterol (P ≤ 0.001), apolipoprotein B (P ≤ 0.01), VLDL-cholesterol, IDL-cholesterol, oxidized-LDL and apolipoprotein B:apolipoprotein A-I ratio (P ≤ 0.05). In MetS patients, urolithin-A only correlated inversely with glucose (P ≤ 0.05). Statin-treated MetS patients with UM-A showed a lipid profile similar to that of healthy normoweight individuals while a poor response to lipid-lowering therapy was observed in MB patients. UMs are potential CMR biomarkers. Overweight-obese individuals with UM-B are at increased risk of cardiometabolic disease, whereas urolithin-A production could protect against CMR factors. Further research is warranted to explore these associations in larger cohorts and whether the effect of lipid-lowering drugs or ellagitannin-consumption on CMR biomarkers depends on individuals' UM. NCT01916239 (https://clinicaltrials.gov/ct2/show/NCT01916239) and ISRCTN36468613 (http://www.isrctn.com/ISRCTN36468613).


Fetal microchimerism in women with breast cancer.
Fetal microchimerism (FMc) describes long-term persistence of small numbers of fetal-derived allogeneic cells in the mother. Although FMc has been implicated as a mechanism of autoimmune disease, it may confer a beneficial effect with immune surveillance of malignant cells. We hypothesized that allogeneic FMc imparts a protective effect against breast cancer. Two observations provided a rationale for the study hypothesis. First, allogeneic cells convey risk reduction for recurrent malignancy in hematopoietic cell transplantation. Second, reduced risk of breast cancer is well recognized among parous compared with nulliparous women. As an initial test of the hypothesis, we investigated 82 women, 35 with breast cancer and 47 who were healthy, for male DNA in peripheral blood, presumed from a prior pregnancy with a male fetus. The prevalence and levels of male DNA were determined by real-time quantitative PCR for the Y chromosome-specific gene DYS14 in DNA extracted from peripheral blood mononuclear cells. FMc was found significantly more often in healthy women than women with breast cancer (43% versus 14%, respectively). Considering the absence of FMc as a risk factor, the odds ratio was 4.4 [95% confidence intervals (95% CI), 1.34-16.99; P = 0.006]. Restricting analysis to women known to had given birth to a son, the odds ratio was 5.9 (95% CI, 1.26-6.69; P = 0.01). Our findings indicate that allogeneic FMc may contribute to reduction in risk of breast cancer. Further studies are indicated and, if confirmed, extended studies to examine whether allogeneic immune surveillance from FMc is deficient in women with breast cancer.


Epigenetic regulation of hematopoietic differentiation by Gfi-1 and Gfi-1b is mediated by the cofactors CoREST and LSD1.
Gfi-1 and Gfi-1b are homologous transcriptional repressors involved in diverse developmental contexts, including hematopoiesis and oncogenesis. Transcriptional repression by Gfi proteins requires the conserved SNAG domain. To elucidate the function of Gfi proteins, we purified Gfi-1b complexes and identified interacting proteins. Prominent among these is the corepressor CoREST, the histone demethylase LSD1, and HDACs 1 and 2. CoREST and LSD1 associate with Gfi-1/1b via the SNAG repression domain. Gfi-1b further recruits these cofactors to the majority of target gene promoters in vivo. Inhibition of CoREST and LSD1 perturbs differentiation of erythroid, megakaryocytic, and granulocytic cells as well as primary erythroid progenitors. LSD1 depletion derepresses Gfi targets in lineage-specific patterns, accompanied by enhanced histone 3 lysine 4 methylation at the respective promoters. Overall, we show that chromatin regulatory proteins CoREST and LSD1 mediate transcriptional repression by Gfi proteins. Lineage-restricted deployment of these cofactors through interaction with Gfi proteins controls hematopoietic differentiation.


Comparative analysis of dose volume histogram reduction algorithms for normal tissue complication probability calculations.
A model for estimating radiotherapy treatment outcome through the probability of damage to normal tissue and the probability of tumour control is a useful tool for treatment plan optimization, dose escalation strategies and other currently used procedures in radiation oncology. Normal tissue complication estimation (NTCP) is here analysed from the point of view of the reliability and internal consistency of the most popular model. Five different dose volume histogram (DVH) reduction algorithms, applied to the Lyman model for NTCP calculation. were analysed and compared. The study was carried out for sets of parameters corresponding to quite different expected dose-response relationships. In particular, we discussed the dependence of the models on the parameters and on the dose bin size in the DVH. The sensitivity of the different reduction schemes to dose inhomogeneities was analysed, using a set of simple DVHs representing typical situations of radiation therapy routine. Significant differences were substantiated between the various reduction methods regarding the sensitivity to the degree of irradiation homogeneity, to the model parameters and to the dose bin size. Structural aspects of the reduction formalism allowed an explanation for these differences. This work shows that DVH reduction for NTCP calculation has still to be considered as a very delicate field and used with extreme care, especially for clinical applications, at least until the actual formulations are tuned against strong clinical data.


Exposure to High Doses of Lipopolysaccharide during Ovalbumin Sensitization Prevents the Development of Allergic Th2 Responses to a Dietary Antigen.
Food allergies are driven by aberrant T helper (Th) 2 cells. Lipopolysaccharide (LPS) influences the development of Th2-mediated diseases, but its role in food allergy and tolerance remains unclear. To address this issue, we established mouse models presenting allergic or tolerant responses to ovalbumin (OVA). Mice sensitized with crude OVA developed Th2 responses including acute diarrhea, increases in serum OVA-specific IgE, dominant production of serum OVA-specific IgG1, increases in Th2-type cytokines and proliferation of mast cells in duodenal and colonic tissues. Sensitization of mice with crude OVA and LPS abrogated Th2-type responses observed in allergic mice. The level of OVA-specific proliferation in mesenteric lymph node CD4(+) T cells was comparable in allergic and tolerant mice, indicating that the tolerance is not caused by anergy and apoptosis of antigen-primed T cells. Expression of Th1- and Th2-type cytokines was suppressed in whole spleen cells and/or purified spleen CD4(+) T cells of tolerant mice, indicating that the tolerance was not caused by the shift from Th2 to Th1. On the other hand, interleukin (IL)-10, a regulatory cytokine produced by regulatory T cells, was upregulated in whole spleen cells and purified spleen CD4(+) T cells of tolerant mice. Furthermore, spleen CD4(+) T cells from tolerant mice suppressed the growth of CD4(+) T cells from DO11.10 mice in co-culture. These results indicate that tolerance is induced in allergic mice by simultaneous exposure to LPS during sensitization with OVA and that a population of T cells producing IL-10 plays an important role in the tolerance induction.


Transcriptional analysis of different stress response genes in Escherichia coli strains subjected to sodium chloride and lactic acid stress.
Survival of Escherichia coli in food depends on its ability to adapt against encountered stress typically involving induction of stress response genes. In this study, the transcriptional induction of selected acid (cadA, speF) and salt (kdpA, proP, proW, otsA, betA) stress response genes was investigated among five E. coli strains, including three Shiga toxin-producing strains, exposed to sodium chloride or lactic acid stress. Transcriptional induction upon lactic acid stress exposure was similar in all but one E. coli strain, which lacked the lysine decarboxylase gene cadA. In response to sodium chloride stress exposure, proW and otsA were similarly induced, while significant differences were observed between the E. coli strains in induction of kdpA, proP and betA. The kdpA and betA genes were significantly induced in four and three strains, respectively, whereas one strain did not induce these genes. The proP gene was only induced in two E. coli strains. Interestingly, transcriptional induction differences in response to sodium chloride stress exposure were associated with survival phenotypes observed for the E. coli strains in cheese as the E. coli strain lacking significant induction in three salt stress response genes investigated also survived poorly compared to the other E. coli strains in cheese.


Effect of posture on plasma immunoreactive atrial natriuretic peptide concentrations in man.
The effect of changes of posture on plasma atrial natriuretic peptide concentrations and renal function was studied in normal human volunteers. Plasma atrial natriuretic peptide concentrations increased in the supine posture, reached a maximum value after 30-60 min, remained elevated for 4 h and decreased to baseline values on return to the upright posture. Inflation of antishock trousers, which apply positive pressure to the legs and lower abdomen, attenuated the fall in plasma atrial natriuretic peptide concentration in the upright position. In the supine posture there were increases in urine flow rate, sodium, lithium, fractional sodium and fractional lithium clearances. Fractional distal water and sodium excretion, and total distal water and sodium reabsorption, which were estimated by the lithium clearance technique, also increased. Heart rate and systolic and diastolic blood pressures decreased in the supine and increased on return to the upright posture. Inflation of antishock trousers prevented the increase in heart rate in the upright posture. The contribution of haemodynamic factors to the increase in plasma atrial natriuretic peptide concentrations in the supine position and the relationship between this increase and the associated changes in renal function are discussed. However, the contribution of atrial natriuretic peptide to these changes is uncertain.


Antitumor efficacy of capecitabine and celecoxib in irradiated and lead-shielded, contralateral human BxPC-3 pancreatic cancer xenografts: clinical implications of abscopal effects.
X-ray therapy (XRT) remains one of the major modalities used to treat patients diagnosed with locally advanced pancreatic adenocarcinoma. However, the effect of XRT on metastatic tumors outside the field of irradiation (abscopal effect) remains largely unknown. In the current study, we examined the effect of XRT alone and in combination with capecitabine and/or celecoxib in both irradiated and lead-shielded contralateral BxPC-3 pancreatic cancer xenografts. This chemoradiation regimen was chosen based on our molecular analysis of pancreatic adenocarcinoma. Athymic mice were injected bilaterally with BxPC-3 cells and treatment was initiated 28 days postimplant. During XRT (2 Gy for 5 consecutive days, administered on days 0 and 24), one flank was irradiated whereas the rest of the body (including the contralateral tumor) was lead shielded. Capecitabine (350 mg/kg) was administered on days 0 to 13 and 24 to 37. Celecoxib was initiated in the diet at 100 ppm (equivalent to 20 mg/kg/d p.o.) and administered throughout the study. In irradiated xenografts, capecitabine and XRT showed synergistic anitiumor efficacy (P=0.008), which was further improved with the addition of celecoxib (P<0.001). In contralateral shielded xenografts, abscopal effects were observed. Whereas monotherapy with XRT showed significant reduction in tumor area in irradiated xenografts, growth was promoted by 23% (P<0.001) in contralateral lead-shielded tumors in the same animals relative to untreated tumors. Interestingly, synergistic antiproliferative efficacy occurred in these contralateral tumors when capecitabine was administered (P<0.001), despite being outside the irradiated field. The addition of celecoxib further inhibited tumor growth (P<0.001). This trimodal combination most effectively stabilized disease in both shielded and irradiated tumors; however, tumor eradication was not observed. There were no significant changes in thymidine phosphorylase, dihydropyrimidine dehydrogenase, or cyclooxygenase-2 mRNA levels in irradiated or lead-shielded tumors, suggesting that efficacy cannot be predicted solely from these previously identified indicators of response. Immunohistochemistry examining the proliferation marker Ki-67 showed concordance with tumor response in both irradiated and contralateral shielded xenografts. These results have implications in the rational design of treatment paradigms for pancreatic cancer where metastatic disease remains the primary cause of patient morbidity and abscopal effects in tumors outside the field of irradiation may affect tumor response.


Involvement of luxS in Biofilm Formation by Capnocytophaga ochracea.
Capnocytophaga ochracea is present in the dental plaque biofilm of patients with periodontitis. Biofilm cells change their phenotype through quorum sensing in response to fluctuations in cell-population density. Quorum sensing is mediated by auto-inducers (AIs). AI-2 is involved in intercellular signaling, and production of its distant precursor is catalyzed by LuxS, an enzyme involved in the activated methyl cycle. Our aim was to clarify the role of LuxS in biofilm formation by C. ochracea. Two luxS-deficient mutants, TmAI2 and LKT7, were constructed from C. ochracea ATCC 27872 by homologous recombination. The mutants produced significantly less AI-2 than the wild type. The growth rates of these mutants were similar to that of the wild-type in both undiluted Tryptic soy broth and 0.5 × Tryptic soy broth. However, according to crystal violet staining, they produced significantly less biofilm than the wild type. Confocal laser scanning microscopy and scanning electron microscopy showed that the biofilm of the TmAI2 strain had a rougher structure than that of the wild type. Complementation of TmAI-2 with extrinsic AI-2 from the culture supernatant of wild-type strain did not restore biofilm formation by the TmAI2 strain, but complementation of LKT7 strain with luxS partially restored biofilm formation. These results indicate that LuxS is involved in biofilm formation by C. ochracea, and that the attenuation of biofilm formation by the mutants is likely caused by a defect in the activated methyl cycle rather than by a loss of AI-2.


Surgery for atrial fibrillation.
The observations made under controlled experimental conditions provided us with a clearer understanding of the mechanisms involved in the generation and perpetuation of atrial fibrillation. The magnitude and rapidity of change that occurs in the activation patterns during atrial fibrillation were not appreciated, however, until the arrhythmia was studied in similar detail in humans. These studies provided the scientific basis for devising a surgical treatment for atrial fibrillation that has been successful in three patients. By converting the atrial fibrillation to normal sinus rhythm, all three of the detrimental sequelae of atrial fibrillation have been alleviated in each of these patients. Despite the fact that these clinical results are preliminary at this point, our experience documents that atrial fibrillation can be cured by surgical means. In the absence of other effective forms of therapy and in view of the devastating complications of the arrhythmia, surgical intervention should be considered a viable option for the treatment of atrial fibrillation.


The impact of planning intervention on business development: Evidence from the Netherlands.
There has been a growing research interest in measuring the impact of planning and land-use regulations on housing market outcomes, but parallel development of the evidence base for the business sector has yet to occur. This article examines the impact of planning intervention on the amount of building investment taking place at sites allocated for industrial and business development. Measures that capture different dimensions of planning intervention are incorporated into models of industrial building investment. The models are estimated using a novel micro dataset on permit activity that covers a sample of industrial and business sites in the Netherlands. The results provide evidence of some of the expected negative effects of the regulatory role of planning intervention, but also show that proactive, targeted planning policies exert a significant and positive influence on investment activity. Specifically, policy-induced improvements to the physical environment will stimulate both new construction and refurbishment activity.


Cell wall teichoic acid glycosylation in Listeria monocytogenes serotype 4b requires gtcA, a novel, serogroup-specific gene.
We have identified a novel gene, gtcA, involved in the decoration of cell wall teichoic acid of Listeria monocytogenes serotype 4b with galactose and glucose. Insertional inactivation of gtcA brought about loss of reactivity with the serotype 4b-specific monoclonal antibody c74.22 and was accompanied by a complete lack of galactose and a marked reduction in the amounts of glucose on teichoic acid. Interestingly, the composition of membrane-associated lipoteichoic acid was not affected. Complementation of the mutants with the cloned gtcA in trans restored galactose and glucose on teichoic acid to wild-type levels. The complemented strains also recovered reactivity with c74.22. Within L. monocytogenes, sequences homologous to gtcA were found in all serogroup 4 isolates but not in strains of any other serotypes. In serotype 4b, gtcA appears to be the first member of a bicistronic operon which includes a gene with homology to Bacillus subtilis rpmE, encoding ribosomal protein L31. In contrast to gtcA, the latter gene appears conserved among all screened serotypes of L. monocytogenes.


Cost-Effectiveness of Cabozantinib in the Second-Line Treatment of Advanced Hepatocellular Carcinoma.
Treatment options are limited for patients with advanced hepatocellular carcinoma (HCC) that progresses after treatment with sorafenib. Cabozantinib, an oral small molecule inhibitor of multiple tyrosine kinase receptors, recently showed improved overall survival (OS) compared with placebo in sorafenib-pretreated patients with advanced HCC in the CELESTIAL trial. This study assessed the cost-effectiveness of cabozantinib for second-line treatment of patients with advanced HCC from a US healthcare system perspective. Cost and utility data were extracted from the CELESTIAL trial and used to determine the cost-effectiveness of cabozantinib compared with placebo plus best supportive care. The main outcome of this study was the incremental cost-effectiveness ratio (ICER), expressed as cost per quality-adjusted life year (QALY) gained by using cabozantinib compared with placebo plus best supportive care in sorafenib-pretreated HCC. In the base-case analysis using data from the CELESTIAL trial, the incremental QALY and ICER were 0.067 and $1,040,675 for cabozantinib compared with placebo and best supportive care. OS reported in the CELESTIAL trial (hazard ratio, 0.76; 95% CI, 0.63-0.92) had the strongest association with the ICER. In one-way sensitivity analyses, there were no scenarios in which cabozantinib was cost-effective. In a cost-threshold analysis, cabozantinib would have to be priced at least $50 per pill to be cost-effective considering a willingness to pay of $100,000 per QALY. Although the CELESTIAL trial demonstrated that cabozantinib improves OS compared with placebo in patients with HCC that progresses after treatment with sorafenib, our analysis shows that cabozantinib is not a cost-effective therapy in this scenario. At current costs, cabozantinib is not cost-effective for second-line therapy of HCC in the United States.


Multivoxel patterns reveal functionally differentiated networks underlying auditory feedback processing of speech.
The everyday act of speaking involves the complex processes of speech motor control. An important component of control is monitoring, detection, and processing of errors when auditory feedback does not correspond to the intended motor gesture. Here we show, using fMRI and converging operations within a multivoxel pattern analysis framework, that this sensorimotor process is supported by functionally differentiated brain networks. During scanning, a real-time speech-tracking system was used to deliver two acoustically different types of distorted auditory feedback or unaltered feedback while human participants were vocalizing monosyllabic words, and to present the same auditory stimuli while participants were passively listening. Whole-brain analysis of neural-pattern similarity revealed three functional networks that were differentially sensitive to distorted auditory feedback during vocalization, compared with during passive listening. One network of regions appears to encode an "error signal" regardless of acoustic features of the error: this network, including right angular gyrus, right supplementary motor area, and bilateral cerebellum, yielded consistent neural patterns across acoustically different, distorted feedback types, only during articulation (not during passive listening). In contrast, a frontotemporal network appears sensitive to the speech features of auditory stimuli during passive listening; this preference for speech features was diminished when the same stimuli were presented as auditory concomitants of vocalization. A third network, showing a distinct functional pattern from the other two, appears to capture aspects of both neural response profiles. Together, our findings suggest that auditory feedback processing during speech motor control may rely on multiple, interactive, functionally differentiated neural systems.


DNA diversity in Hawaiian endemic plant Schiedea globosa.
This is the first report of a study devoted to the population genetics of speciation in the endemic Hawaiian plant genus Schiedea (Caryophyllaceae). Here, we report the estimates of DNA sequence diversity and divergence in a newly isolated nuclear gene from Maui and Oahu Schiedea globosa populations. Overall, the species-wide average heterozygosity per silent site is pi = 0.3%. The silent DNA diversity on the older island of Oahu (pi = 0.24%) is almost twice as high as on the younger Maui (pi = 0.14%). Consistent with this, the haplotype phylogeny suggests a more recent origin of the Maui populations. There is no significant isolation between the two Maui populations (F(st)=0.027), while isolation between the two islands is high (F(st)=0.57, P<0.0001). Pairwise mismatch distributions suggest population growth approximately 660 and 310 thousand generations ago for the Oahu and the Maui populations, respectively, which may be the minimal age for these populations. This is consistent with a fairly neutral frequency spectrum (Tajima's D is 0.34 and -0.94 for the Oahu and the Maui populations, respectively), suggesting that both populations are sufficiently old to have recovered from any initial founder effects. Relatively high nuclear DNA diversity in the S. globosa populations illustrates the usefulness of a DNA sequence-based approach to the population genetics of island plant populations.


Social and environmental malaria risk factors in urban areas of Ouagadougou, Burkina Faso.
Despite low endemicity, malaria remains a major health problem in urban areas where a high proportion of fevers are presumptively treated using anti-malarial drugs. Low acquired malaria immunity, behaviour of city-dwellers, access to health care and preventive interventions, and heterogenic suitability of urban ecosystems for malaria transmission contribute to the complexity of the malaria epidemiology in urban areas. The study was designed to identify the determinants of malaria transmission estimated by the prevalence of anti-circumsporozoite (CSP) antibodies, the prevalence and density of Plasmodium falciparum infection, and the prevalence of malarial disease in areas of Ouagadougou, Burkina-Faso. Thick blood smears, dried blood spots and clinical status have been collected from 3,354 randomly chosen children aged 6 months to 12 years using two cross-sectional surveys (during the dry and rainy seasons) in eight areas from four ecological strata defined according to building density and land tenure (regular versus irregular). Demographic characteristics, socio-economic information, and sanitary and environmental data concerning the children or their households were simultaneously collected. Dependent variables were analysed using mixed multivariable models with random effects, taking into account the clustering of participants within compounds and areas. Overall prevalences of CSP-antibodies and P. falciparum infections were 7.7% and 16.6% during the dry season, and 12.4% and 26.1% during the rainy season, respectively, with significant differences according to ecological strata. Malaria risk was significantly higher among children who i) lived in households with lower economic or education levels, iii) near the hydrographic network, iv) in sparsely built-up areas, v) in irregularly built areas, vi) who did not use a bed net, vii) were sampled during the rainy season or ii) had traveled outside of Ouagadougou. Malaria control should be focused in areas which are irregularly or sparsely built-up or near the hydrographic network. Furthermore, urban children would benefit from preventive interventions (e.g. anti-vectorial devices or chemoprophylaxis) aimed at reducing malaria risk during and after travel in rural areas.


Editorial: The continuing contributions of multimodal treatment of attention over nearly two decades to initial attention-deficit hyperactivity disorder pharmacotherapy and long-term clinical course.
The initial results of the Multimodal Treatment of Attention Deficit Hyperactivity Disorder (MTA) trial had profound effects on the way that Attention-Deficit Hyperactivity Disorder (ADHD) is managed clinically. Children from the original MTA cohort as well as a control group have been followed longitudinally for well over a decade and are beginning to provide important data regarding the long-term clinical course, treatment and consequences of ADHD into adulthood. Two articles in this issue of JCPP highlight important contributions from the MTA cohort. Swanson et al. highlights the potential long-term effects of stimulants on height whereas Sibley et al. highlights the importance of using multiple informants in assessing adulthood ADHD symptoms similar to children and suggest that current DSM criteria for ADHD may be overly stringent.


The use of organotypic hippocampal slice cultures to evaluate protection by non-competitive NMDA receptor antagonists against excitotoxicity.
There is great interest in testing neuroprotectants which inhibit the neurodegeneration that results from excessive activation of N-methyl-D-aspartate (NMDA) receptors. As an alternative to in vivo testing in animal models, we demonstrate here the use of a complex in vitro model to compare the efficacy and toxicity of NMDA receptor inhibitors. Organotypic hippocampal slice cultures were used to compare the effectiveness of the Alzheimer's disease drug, memantine, the Parkinson's disease drug, procyclidine, and the novel neuroprotectant, gacyclidine (GK11), against NMDA-induced toxicity. All three drugs are non-competitive NMDA receptor open-channel blockers that inhibit excitotoxic injury, and their neuroprotective capacities have been extensively investigated in vivo in animal models. They have also been evaluated as potential countermeasure agents against organophosphate poisoning. Quantitative densitometric image analysis of propidium iodide uptake in hippocampal regions CA1, CA3 and DG, showed that, after exposure to 10microM NMDA for 24 hours, GK11 was the most potent of the three drugs, with an IC50 of about 50nM and complete protection at 250nM. When applied at high doses, GK11 was still the more potent neuroprotectant, and also the least cytotoxic. These findings are consistent with those from in vivo tests in rodents. We conclude that the slice culture model provides valuable pre-clinical data, and that applying the model to the screening of neuroprotectants might significantly limit the use of in vivo tests in animals.


Is the metabolic syndrome caused by a high fructose, and relatively low fat, low cholesterol diet?
The metabolic syndrome (MetS) is manifested by a lipid triad which includes elevated serum triglycerides, small LDL particles, and low high-density lipoprotein (HDL) cholesterol, by central obesity (central adiposity), insulin resistance, glucose intolerance and elevated blood pressure, and it is associated with an increased risk of type 2 diabetes and coronary heart disease. We have developed a new hypothesis regarding MetS as a consequence of a high intake in carbohydrates and food with a high glycemic index, particularly fructose, and relatively low intake of cholesterol and saturated fat. We support our arguments through animal studies which have shown that exposure of the liver to increased quantities of fructose leads to rapid stimulation of lipogenesis and accumulation of triglycerides. The adipocytes store triglycerides in lipid droplets, leading to adipocyte hypertrophy. Adipocyte hypertrophy is associated with macrophage accumulation in adipose tissue. An important modulator of obesity-associated macrophage responses in white adipose tissue is the death of adipocytes. Excess exposure to fructose intake determines the liver to metabolize high doses of fructose, producing increased levels of fructose end products, like glyceraldehyde and dihydroxyacetone phosphate, that can converge with the glycolytic pathway. Fructose also leads to increased levels of advanced glycation end products. The macrophages exposed to advanced glycation end products become dysfunctional and, on entry into the artery wall, contribute to plaque formation and thrombosis.


Recommendations for the use of transcranial Doppler ultrasonography to determine the existence of cerebral circulatory arrest as diagnostic support for brain death.
The clinical criteria for brain death consist of the demonstration of the absence of any clinical sign of encephalic activity. Confirmatory testing is usually not required for the diagnosis of brain death, except in some special situations that Spanish law details. In these situations demonstrating cerebral circulatory arrest (CCA) by cerebral flow studies is necessary to support the diagnosis of brain death. To review the use of transcranial Doppler ultrasonography (TCD) for confirming brain death and to establish uniform criteria for the routine use of TCD as a confirmatory test. Based on literature analysis, the authors developed guidelines for performance and interpretation of TCD in clinically brain-dead patients, in order to confirm the diagnosis. The active members of the Spanish Neurosonology Society (SONES) reviewed an initial draft, until a consensus was reached. In a clinically brain-dead patient, specific intracranial flow patterns indicating CCA can be visualized by TCD. The specific flow patterns are the presence of reverberating flow and/or systolic spikes, and should be detected in both middle cerebral arteries and also in the basilar artery. We recommend repeating the examination within 30 minutes to confirm the findings. TCD is a useful method for detecting CCA and therefore can be used to confirm brain death in a clinically brain-dead patient. The presence of reverberating flow, systolic spikes or absence of flow in the basilar and both middle cerebral arteries observed in two examinations is highly specific for the prediction of CCA and brain death in all patients.


Antitumor activity of the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor gefitinib (ZD1839, Iressa) in non-small cell lung cancer cell lines correlates with gene copy number and EGFR mutations but not EGFR protein levels.
Recognition that the epidermal growth factor receptor (EGFR) was a therapeutic target in non-small cell lung cancer (NSCLC) and other cancers led to development of the small-molecule receptor tyrosine kinase inhibitors gefitinib and erlotinib. Clinical trials established that EGFR tyrosine kinase inhibitors produced objective responses in a minority of NSCLC patients. We examined the sensitivity of 23 NSCLC lines with wild-type or mutated EGFR to gefitinib to determine genes/proteins related to sensitivity, including EGFR and HER2 cell surface expression, phosphorylated EGFR expression, EGFR gene copy number, and EGFR mutational status. Downstream cell cycle and signaling events were compared with growth-inhibitory effects. We determined gefitinib sensitivity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, EGFR expression by fluorescence-activated cell sorting and immunohistochemistry, phosphorylated EGFR by Western blotting, EGFR gene copy number by fluorescence in situ hybridization, and EGFR mutation by sequencing. The cellular effects of gefitinib on cell cycle were determined by flow cytometry and the molecular effects of gefitinib EGFR inhibition on downstream signal proteins by Western blotting. Gefitinib in vivo effects were evaluated in athymic nude mice bearing sensitive and resistant NSCLC xenografts. There was a significant correlation between EGFR gene copy number, EGFR gene mutations, and gefitinib sensitivity. EGFR protein was necessary but not sufficient for predicting sensitivity. Gefitinib-sensitive lines showed a G(1) cell cycle arrest and inactivation of downstream signaling proteins; resistant cell lines had no changes. The in vivo effects mirrored the in vitro effects. This panel of NSCLC lines characterized for gefitinib response was used to identify predictive molecular markers of response to gefitinib. Several of these have subsequently been shown to identify NSCLC patients likely to benefit from gefitinib therapy.


Treponema parvum sp. nov., a small, glucoronic or galacturonic acid-dependent oral spirochaete from lesions of human periodontitis and acute necrotizing ulcerative gingivitis.
Small oral spirochaetes with a strict dependence on either glucuronic acid (GluA) or galacturonic acid (GalA) were isolated from European patients with periodontitis and from Chinese patients with either gingivitis or acute necrotizing ulcerative gingivitis (ANUG). Thirteen such isolates were similar phenotypically to Treponema pectinovorum ATCC 33768T and this classification was confirmed by 16S rRNA sequencing. However, four isolates differed from T. pectinovorum by their small cell size, by a prominent beta-glucuronidase activity, by a distinct protein and antigen profile, by an inability to grow on pectin as sole source of carbohydrate and by a markedly enhanced growth rate when supplied with a second carbohydrate (L-arabinose, D-galactose, D-glucose, D-fructose, D-mannitol, D-mannose, pectin, D-ribose or D-xylose) in addition to the essential GluA/GalA. By 16S rRNA sequencing these four isolates clustered in the recently described phylotype 'Smibert-2'. T. pectinovorum (14 strains) and 'Smibert-2' (four isolates with beta-glucuronidase activity) could each be subdivided into two serotypes based on immunoblot reactivity with two mAbs. Representatives of the two groups, including T. pectinovorum ATCC 33768T, showed a 1:2:1-type periplasmic flagellar arrangement. 'Smibert-2' is described as a novel species, Treponema parvum sp. nov., with isolate OMZ 833T (= ATCC 700770T) proposed as the type strain and OMZ 842 (= ATCC 700773) as reference strain for a second serotype.


A microstructurally driven model for pulmonary artery tissue.
A new constitutive model for elastic, proximal pulmonary artery tissue is presented here, called the total crimped fiber model. This model is based on the material and microstructural properties of the two main, passive, load-bearing components of the artery wall, elastin, and collagen. Elastin matrix proteins are modeled with an orthotropic neo-Hookean material. High stretch behavior is governed by an orthotropic crimped fiber material modeled as a planar sinusoidal linear elastic beam, which represents collagen fiber deformations. Collagen-dependent artery orthotropy is defined by a structure tensor representing the effective orientation distribution of collagen fiber bundles. Therefore, every parameter of the total crimped fiber model is correlated with either a physiologic structure or geometry or is a mechanically measured material property of the composite tissue. Further, by incorporating elastin orthotropy, this model better represents the mechanics of arterial tissue deformation. These advancements result in a microstructural total crimped fiber model of pulmonary artery tissue mechanics, which demonstrates good quality of fit and flexibility for modeling varied mechanical behaviors encountered in disease states.


Non-vitamin K antagonist oral anticoagulants (NOACs) post-percutaneous coronary intervention: a network meta-analysis.
Clinicians must balance the risks of bleeding and thrombosis after percutaneous coronary intervention (PCI) in people with an indication for anticoagulation. The potential of non-vitamin K antagonists (NOACs) to prevent bleeding complications is promising, but evidence remains limited. To review the evidence from randomised controlled trials assessing the efficacy and safety of non-vitamin K antagonist oral anticoagulants (NOACs) compared to vitamin K antagonists post-percutaneous coronary intervention (PCI) in people with an indication for anticoagulation. We identified studies by searching CENTRAL, MEDLINE, Embase, the Conference Proceedings Citation Index - Science and two clinical trials registers in February 2019. We checked bibliographies of identified studies and applied no language restrictions. We searched for randomised controlled trials (RCT) that compared NOACs and vitamin K antagonists for people with an indication for anticoagulation who underwent PCI. Two review authors independently checked the results of searches to identify relevant studies, assessed each included study, and extracted study data. We conducted random-effects, pairwise analyses using Review Manager 5 and network meta-analyses (NMA) using the R package 'netmeta'. We ranked competing treatments by P scores, which are derived from the P values of all pairwise comparisons, and allow ranking of treatments on a continuous 0 to 1 scale. We identified nine RCTs that met the inclusion criteria, but four were ongoing trials, and were not included in this analysis. We included five RCTs, with 8373 participants, in the NMA (two RCTs compared apixaban to a vitamin K antagonist, two RCTs compared rivaroxaban to a vitamin K antagonist, and one RCT compared dabigatran to a vitamin K antagonist). Very low- to moderate-certainty evidence suggests little or no difference between NOACs and vitamin K antagonists in death from cardiovascular causes (not reported in the dabigatran trial), myocardial infarction, stroke, death from any cause, and stent thrombosis. Apixaban (RR 0.85, 95% CI 0.77 to 0.95), high dose rivaroxaban (RR 0.86, 95% CI 0.74 to 1.00), and low dose rivaroxaban (RR 0.80, 95% CI 0.68 to 0.92) probably reduce the risk of recurrent hospitalisation compared with vitamin K antagonists. No studies looked at health-related quality of life. Very low- to moderate-certainty evidence suggests that NOACs may be safer than vitamin K antagonists in terms of bleeding. Both high dose dabigatran (RR 0.53, 95% CI 0.29 to 0.97), and low dose dabigatran (RR 0.38, 95% CI 0.21 to 0.70) may reduce major bleeding more than vitamin K antagonists. High dose dabigatran (RR 0.83, 95% CI 0.72 to 0.96), low dose dabigatran (RR 0.66, 95% CI 0.58 to 0.75), apixaban (RR 0,67 , 95% Cl 0.51 to 0.88), high dose rivaroxaban (RR 0.66, 95% CI 0.52 to 0.83), and low dose rivaroxaban (RR 0.71, 95% CI 0.57 to 0.88) probably reduce non-major bleeding more than vitamin K antagonists. The results from the NMA were inconclusive between the different NOACs for all primary and secondary outcomes. Very low- to moderate-certainty evidence suggests no meaningful difference in efficacy outcomes between non-vitamin K antagonist oral anticoagulants (NOAC) and vitamin K antagonists following percutaneous coronary interventions (PCI) in people with non-valvular atrial fibrillation. NOACs probably reduce the risk of recurrent hospitalisation for adverse events compared with vitamin K antagonists. Low- to moderate-certainty evidence suggests that dabigatran may reduce the rates of major and non-major bleeding, and apixaban and rivaroxaban probably reduce the rates of non-major bleeding compared with vitamin K antagonists. Our network meta-analysis did not show superiority of one NOAC over another for any of the outcomes. Head to head trials, directly comparing NOACs against each other, are required to provide more certain evidence.


Subtypes of progressive aphasia: application of the International Consensus Criteria and validation using β-amyloid imaging.
Primary progressive aphasia comprises a heterogeneous group of neurodegenerative conditions with diverse clinical profiles and underlying pathological substrates. A major development has been the publication of the recent International Consensus Criteria for the three major variants namely: semantic, non-fluent/agrammatic and logopenic. The logopenic variant is assumed to represent an atypical presentation of Alzheimer pathology although evidence for this is, at present, limited. The semantic and non-fluent/agrammatic variants are largely associated with frontotemporal lobar degeneration with TDP-43 and tau pathology, respectively. The applicability of the International Consensus Criteria to an unselected clinical sample is unknown and no agreed clinical evaluation scale on which to derive the diagnosis exists. We assessed 47 consecutive cases of primary progressive aphasic seen over a 3-year period in a specialist centre, using a newly developed progressive aphasia language scale. A subgroup of 30 cases underwent (11)C-labelled Pittsburgh Compound B positron emission tomography imaging, a putative biomarker of Alzheimer's disease that detects β-amyloid accumulation, and they were compared with an age-matched group (n = 10) with typical, predominately amnestic Alzheimer's disease. The application of an algorithm based on four key speech and language variables (motor speech disorders, agrammatism, single-word comprehension and sentence repetition) classified 45 of 47 (96%) of patients and showed high concordance with the gold standard expert clinical diagnosis based on the International Consensus Criteria. The level of neocortical β-amyloid burden varied considerably across aphasic variants. Of 13 logopenic patients, 12 (92%) had positive β-amyloid uptake. In contrast, one of nine (11%) semantic variant and two of eight (25%) non-fluent/agrammatic cases were positive. The distribution of β-amyloid across cortical regions of interest was identical in cases with the logopenic variant to that of patients with typical Alzheimer's disease although the total load was lower in the aphasic cases. Impairments of sentence repetition and sentence comprehension were positively correlated with neocortical burden of β-amyloid, whereas impaired single-word comprehension showed a negative correlation. The International Consensus Criteria can be applied to the majority of cases with primary progressive aphasic using a simple speech and language assessment scale based upon four key variables. β-amyloid imaging confirms the higher rate of Alzheimer pathology in the logopenic variant and, in turn, the low rates in the other two variants. The study offers insight into the biological basis of clinical manifestations of Alzheimer's disease, which appear topographically independent of β-amyloid load.


Regime shifts, thresholds and multiple stable states in freshwater ecosystems; a critical appraisal of the evidence.
The concepts of ecosystem regime shifts, thresholds and alternative or multiple stable states are used extensively in the ecological and environmental management literature. When applied to aquatic ecosystems, these terms are used inconsistently reflecting differing levels of supporting evidence among ecosystem types. Although many aquatic ecosystems around the world have become degraded, the magnitude and causes of changes, relative to the range of historical variability, are poorly known. A working group supported by the Australian Centre for Ecological Analysis and Synthesis (ACEAS) reviewed 135 papers on freshwater ecosystems to assess the evidence for pressure-induced non-linear changes in freshwater ecosystems; these papers used terms indicating sudden and non-linear change in their titles and key words, and so was a positively biased sample. We scrutinized papers for study context and methods, ecosystem characteristics and focus, types of pressures and ecological responses considered, and the type of change reported (i.e., gradual, non-linear, hysteretic or irreversible change). There was little empirical evidence for regime shifts and changes between multiple or alternative stable states in these studies although some shifts between turbid phytoplankton-dominated states and clear-water, macrophyte-dominated states were reported in shallow lakes in temperate climates. We found limited understanding of the subtleties of the relevant theoretical concepts and encountered few mechanistic studies that investigated or identified cause-and-effect relationships between ecological responses and nominal pressures. Our results mirror those of reviews for estuarine, nearshore and marine aquatic ecosystems, demonstrating that although the concepts of regime shifts and alternative stable states have become prominent in the scientific and management literature, their empirical underpinning is weak outside of a specific environmental setting. The application of these concepts in future research and management applications should include evidence on the mechanistic links between pressures and consequent ecological change. Explicit consideration should also be given to whether observed temporal dynamics represent variation along a continuum rather than categorically different states.


Repeated exposure to vicarious pain alters electrocortical processing of pain expressions.
Repeated exposure to others in pain has been shown to bias vicarious pain perception, but the neural correlates of this effect are currently not known. The current study therefore aimed at measuring electrocortical responses to facial expressions of pain following exposure to expressions of pain. To this end, a between-subject design was adopted. Participants in the Exposure group were exposed to facial expressions of intense pain, while the participants in the Control group were exposed to neutral expressions before performing the same pain detection task. As in previous studies, participants in the Exposure group showed a significantly more conservative bias when judging facial expressions pain, meaning that they were less inclined to judge moderate pain expressions as painful compared to participants in the Control group. Event-related potential analyses in response to pain or neutral expressions indicated that this effect was related to a relative decrease in the central late positive potential responses to pain expressions. Furthermore, while the early N170 response was not influenced by repeated exposure to pain expressions, the P100 component showed an adaptation effect in the Control group only. These results suggest that repeated exposure to vicarious pain do not influence early event-related potential responses to pain expressions but decreases the late central positive potential. These results are discussed in terms of changes in the perceived saliency of pain expressions following repeated exposure.


Pancreaticoduodenectomy with or without extended retroperitoneal lymphadenectomy for periampullary adenocarcinoma: comparison of morbidity and mortality and short-term outcome.
This prospective, randomized, single-institution trial was designed to evaluate the end points of mortality, morbidity, and survival in patients undergoing standard versus radical (extended) pancreaticoduodenectomy (including distal gastrectomy and retroperitoneal lymphadenectomy). Numerous retrospective reports and one prospective randomized trial have suggested that the performance of an extended lymphadenectomy in association with a pancreaticoduodenal resection may improve long-term survival for some patients with pancreatic and other periampullary adenocarcinomas. Many of these previously published studies can be criticized for their retrospective and nonrandomized designs, for the inclusion of nonconcurrent control groups, and for their small numbers. Between April 1996 and December 1997, 114 patients with periampullary adenocarcinoma were enrolled in an ongoing, prospective, randomized trial at The Johns Hopkins Hospital. After intraoperative verification of completely resected periampullary adenocarcinoma, the patients were randomized to receive either a standard pancreaticoduodenectomy (removing only the peripancreatic lymph nodes en bloc with the specimen) or a radical pancreaticoduodenectomy (standard resection plus distal gastrectomy and retroperitoneal lymphadenectomy). All pathology specimens were reviewed and categorized. The postoperative morbidity, mortality, and short-term outcomes were examined. Of the 114 patients randomized, 56 underwent a standard pancreaticoduodenectomy and 58 a radical pancreaticoduodenectomy. The two groups were statistically similar with regard to age and gender, but there was a higher percentage of white patients in the radical group. All the patients in the radical group underwent distal gastric resection, whereas 86% of the patients in the standard group underwent pylorus preservation. The mean operative time in the radical group was 6.8 hours, compared with 6.2 hours in the standard group. There were no significant differences between the two groups with respect to the intraoperative blood loss, transfusion requirements, location of primary tumor, mean tumor size, positive lymph node status, or positive margin status. There were three deaths in the standard group and two in the radical group. The complication rates were 34% for the standard group and 40% for the radical group. Patients undergoing radical resection had a higher incidence of early delayed gastric emptying but had similar rates of other complications, such as pancreatic fistula, wound infection, intraabdominal abscess, and need for reoperation. The mean total number of lymph nodes resected was higher in the radical group. Of the 58 patients in the radical group, only 10% had metastatic carcinoma in the resected retroperitoneal lymph nodes, and none of those patients had the retroperitoneal nodes as the only site of lymph node involvement. The 1-year actuarial survival rate for patients surviving the immediate postoperative periods was 77% for the standard resection group and 83% for the radical resection group. These data demonstrate that radical pancreaticoduodenectomy (with the addition of a distal gastrectomy and extended retroperitoneal lymphadenectomy to a standard pancreaticoduodenectomy) can be performed with similar morbidity and mortality to standard pancreaticoduodenectomy. However, the survival data are not sufficiently mature and the numbers of patients enrolled are not adequate to allow firm conclusions to be drawn regarding survival benefit.


To use or not to use. What influences why women veterans choose VA health care.
Effects of advances in Department of Veterans Affairs (VA) women's health care on women veterans' health care decision making are unknown. Our objective was to determine why women veterans use or do not use VA health care. Cross-sectional survey of 2,174 women veteran VA users and VA-eligible nonusers throughout southern California and southern Nevada. VA utilization, attitudes toward care, and socio-demographics. Reasons cited for VA use included affordability (67.9%); women's health clinic (WHC) availability (58.8%); quality of care (54.8%); and convenience (47.9%). Reasons for choosing health care in non-VA settings included having insurance (71.0%); greater convenience of non-VA care (66.9%); lack of knowledge of VA eligibility and services (48.5%); and perceived better non-VA quality (34.5%). After adjustment for socio-demographics, health characteristics, and VA priority group, knowledge deficits about VA eligibility and services and perceived worse VA care quality predicted outside health care use. VA users were less likely than non-VA users to have after-hours access to nonemergency care, but more likely to receive both general and gender-related care from the same clinic or provider, to use a WHC for gender-related care, and to consider WHC availability very important. Lack of information about VA, perceptions of VA quality, and inconvenience of VA care, are deterrents to VA use for many women veterans. VA WHCs may foster VA use. Educational campaigns are needed to fill the knowledge gap regarding women veterans' VA eligibility and advances in VA quality of care, while VA managers consider solutions to after-hours access barriers.


Capitalize on dimensionality increasing techniques for improving Face Recognition Grand Challenge performance.
This paper presents a novel pattern recognition framework by capitalizing on dimensionality increasing techniques. In particular, the framework integrates Gabor image representation, a novel multiclass Kernel Fisher Analysis (KFA) method, and fractional power polynomial models for improving pattern recognition performance. Gabor image representation, which increases dimensionality by incorporating Gabor filters with different scales and orientations, is characterized by spatial frequency, spatial locality, and orientational selectivity for coping with image variabilities such as illumination variations. The KFA method first performs nonlinear mapping from the input space to a high-dimensional feature space, and then implements the multiclass Fisher discriminant analysis in the feature space. The significance of the nonlinear mapping is that it increases the discriminating power of the KFA method, which is linear in the feature space but nonlinear in the input space. The novelty of the KFA method comes from the fact that 1) it extends the two-class kernel Fisher methods by addressing multiclass pattern classification problems and 2) it improves upon the traditional Generalized Discriminant Analysis (GDA) method by deriving a unique solution (compared to the GDA solution, which is not unique). The fractional power polynomial models further improve performance of the proposed pattern recognition framework. Experiments on face recognition using both the FERET database and the FRGC (Face Recognition Grand Challenge) databases show the feasibility of the proposed framework. In particular, experimental results using the FERET database show that the KFA method performs better than the GDA method and the fractional power polynomial models help both the KFA method and the GDA method improve their face recognition performance. Experimental results using the FRGC databases show that the proposed pattern recognition framework improves face recognition performance upon the BEE baseline algorithm and the LDA-based baseline algorithm by large margins.


Unbiased analysis by high throughput sequencing of the viral diversity in fetal bovine serum and trypsin used in cell culture.
Fetal bovine serum (FBS) and trypsin are reagents used in cell culture and have been the source of viral contamination of pharmaceutical products. We performed high throughput sequencing (HTS) of two pools of commercial batches of FBS and three commercial batches of trypsin. Taxonomies were assigned by comparing sequences of contigs and singletons to the entire NCBI nucleic acid and protein databases. The same major viral species were evidenced between batches of a given reagent but the proportion of viral reads among total reads varied markedly between samples (from 0.002% to 22.7%). In FBS, the sequences found were mainly from bovine viral diarrhea virus (BVDV) 1 to 3 and bovine parvovirus 3 (BPV3). The BVDV sequences derived from FBS showed only minor discrepancies with primers generally used for the screening of BVDV. Viral sequences in trypsin were mainly from porcine circovirus type 2. Other known viral sequences at lower read counts and potential new viral species (bovine parvovirus and bovine pegivirus) were evidenced. The load of some known and new viruses detected by HTS could be quantified by qPCR. Results of HTS provide a framework for evaluating the pertinence of control measures including the design of PCRs, bioassays and inactivation procedures.