Intention of parents to have male children vaccinated with the human papillomavirus vaccine.
Although already approved for use in males in some jurisdictions, there is little information about parental attitudes toward having their sons receive the human papillomavirus (HPV) vaccine. The goal of this study was to ascertain parental intentions to vaccinate their sons with an HPV vaccine and to determine factors that predict this intention. Parents of children aged 8-18 years were recruited from across Canada through random digit dialling. Participants were asked to respond to a series of questions in the context of a Grade 6 (age 11/12 years old), publicly funded school-based HPV vaccine programme, including their intention to vaccinate their sons with the HPV vaccine. Parents were also asked about a series of characteristics thought to predict intention to vaccinate as well as demographic characteristics. Backwards logistic regression was conducted to calculate adjusted odds ratios (AOR) to identify the factors that are predictive of parents' intention to vaccinate their son(s) against HPV. Of the 1381 respondents with male children, 67.8% (95% CI 65.3 to 70.3) intend to vaccinate their son(s) against HPV. Parents who had positive attitudes toward vaccines and the HPV vaccine in particular (AOR 41.5, 95% CI 9.5 to 181.7), parents who were influenced by subjective norms (AOR 7.8, 95% CI 5.8 to 10.5), parents who felt that the vaccine had limited influence on sexual behaviour (AOR 2.3, 95% CI 1.6 to 3.3) and parents who were aware of HPV (AOR 1.4, 95% CI 1.1 to 2.0) were significantly more likely to report an intention to vaccinate boys against HPV. In contrast, residence in British Columbia compared to Atlantic Canada (AOR 0.4, 95% CI 0.2 to 0.8) and higher education (AOR 0.7, 95% CI 0.5 to 0.9) were negatively associated with intention to vaccinate. Parents who reported an intention to vaccinate their daughters were also highly likely to report an intention to vaccinate their sons (kappa = 0.9, p<0.001). The majority of Canadian parents would intend to have their male children receive the HPV vaccine in the context of a publicly funded school-based immunisation programme. Overall attitudes toward vaccine, recommendations from health professionals and impact of the vaccine on sexual practices are important predictors of intention to have a male child receive the HPV vaccine.


The Voicemail Elicitation Task: Functional Workplace Language Assessment for Persons With Traumatic Brain Injury.
Purpose Politeness markers (PMs) are words that enhance cooperativity in dialogue and are an essential component of professional/work communication. Persons with moderate/severe traumatic brain injury (TBI) underuse PMs in connected speech and have employment stability issues. The voicemail elicitation task (VET) is a standardized computerized language sampling procedure measuring PM rate in role-play tasks. Our purpose is to provide preliminary data establishing the potential of a screening assessment tool for professional/work communication. Method We measured VET performance using spoken PMs per minute (PMpM). We present data from 63 persons. Forty-three participants with TBI (22-65 years old, ≥ 1-year postinjury) worked in midlevel jobs before their injury and attempted work return after injury at the same job level. Twenty participants with TBI did not maintain work > 1 year (unstably employed), and 23 did maintain work for ≥ 1 year (stably employed). Twenty controls without history of neurological impairment working at the same job level also completed the VET protocol. We analyzed the data using between-group comparison with 1-way analysis of variance and post hoc analysis. We used receiver operating characteristic curve analysis to calculate sensitivity and specificity, as well as an optimal cutoff value for a screening measure. Results Group differences, F(2, 60) = 19.59, p = .0001, η2 = .376, were identified between unstably employed persons with TBI performing with lower PMpM scores than the stably employed TBI group and the control group. Receiver operating characteristic curve analysis indicated a cutoff score of 11.55 PMpM. There was acceptable specificity (0.700, 95% CI [0.499, 0.901]) and sensitivity (0.696, 95% CI [0.508, 0.883]) for a screening tool indicating further assessment of social communication. Conclusion The VET holds promise as a clinical screening tool to identify persons at risk for social communication-related job instability after TBI and the need for a more comprehensive social communication assessment.


Global warming will bring new fungal diseases for mammals.
Fungi are major pathogens of plants, other fungi, rotifers, insects, and amphibians, but relatively few cause disease in mammals. Fungi became important human pathogens only in the late 20th century, primarily in hosts with impaired immunity as a consequence of medical interventions or HIV infection. The relatively high resistance of mammals has been attributed to a combination of a complex immune system and endothermy. Mammals maintain high body temperatures relative to environmental temperatures, creating a thermally restrictive ambient for the majority of fungi. According to this view, protection given by endothermy requires a temperature gradient between those of mammals and the environment. We hypothesize that global warming will increase the prevalence of fungal diseases in mammals by two mechanisms: (i) increasing the geographic range of currently pathogenic species and (ii) selecting for adaptive thermotolerance for species with significant pathogenic potential but currently not pathogenic by virtue of being restricted by mammalian temperatures.


Public policy implications of tobacco industry smuggling through Native American reservations into Canada.
From 1980 to 1994, the Canadian government enacted major tax increases on tobacco products. These actions initiated significant tobacco smuggling from the United States into Canada through a few U.S. Native American reservations to undercut the price of Canadian tobacco products. The tobacco industry blamed rampant smuggling on excessive taxation; however, research shows that the tobacco industry had actually promoted smuggling schemes to both increase profits and provide an argument for tobacco taxation reduction. Although the smuggling has resulted in numerous U.S. and Canadian criminal convictions of tobacco industry officials and partners, significant smuggling continues throughout the world. For the few Native Americans involved, the smuggling was lucrative and they were able to avoid criminal prosecution through tribal sovereignty. Industry-supported tobacco smuggling has had a profoundly negative effect on Canadian public health that must be brought to light to prevent future similar occurrences.


The impact of the new biology on radiation risks in space.
Radiation is considered to be one of three or four major hazards for personnel in space and has emerged as the most critical issue to be resolved for long-term missions, both orbital and interplanetary. Space habitats are stressful and dangerous environments. Health and medical consequences arising from microgravity, stress, and trauma include weakened immune systems, increased viral activity, and loss of bone mass. The greatest risks from radiation are generally assumed to be cancers and possibly damage to the central nervous system. Synergistic effects arising from the other environmental hazards along with abscopal and exogenic factors are likely. Space programs represent an exceptional opportunity for examining the biological consequences of low-dose exposures of humans to radiation at every level of progression. Although astronauts are a relatively small population, they are healthy, physically active volunteers who undergo extensive testing and medical examinations before, during, and after protracted exposures with periodic follow-up examinations. The radiation environments along with other hazards are likewise monitored and documented. Extensive international research programs are in progress. Seven years ago the U.S. National Aeronautics and Space Administration established the National Space Biomedical Research Institute through a cooperative agreement with a consortium of research and academic institutions in order to address radiation issues through a concerted, programmatic effort. Advanced technologies are rapidly being incorporated into these programs to determine the significance of new biological data and to evaluate the interplay among the different medical hazards. Programmatic in vivo and in vitro studies of the processes leading to carcinogenesis are in progress. Drugs and dietary supplements are being examined at the cellular and in vivo levels to assess their potential as dose-modifying agents. The infrastructure of this new approach, recent results, and research in progress are reviewed and discussed.


Modeling ultrafast exciton deactivation in oligothiophenes via nonadiabatic dynamics.
Ultrafast excited-state processes play a key role in organic electronics and photovoltaics, governing the way of how excitons can relax and separate. Through the use of nonadiabatic excited-state dynamics, relaxation processes were investigated at the sub-picosecond timescale in thiophene and oligothiophenes (nT, n = 2, 3, 4), prototype oligomers for efficient π-electron conjugated polymers adopted in photovoltaics. For thiophene, TDDFT and TDA nonadiabatic excited-state dynamics revealed ultrafast nonradiative relaxation processes through ring opening and ring puckering, bringing the system to an S1/S0 conical intersection seam. The computed relaxation time is 110 fs, matching well the experimental one (∼105 fs). In oligothiophenes (n = 2-4), high-energy (hot) excitations were considered. Exciton relaxation through the manifold of excited states to the lowest excited state is predicted to occur within ∼150-200 fs, involving bond stretching, ring puckering, and torsional oscillations. For the longer oligomer (4T), the ultrafast relaxation process leads to exciton localization over three thiophene rings in 150 fs. These data agree with the self-localization mechanism (∼100-200 fs) observed for poly(3-hexylthiophene) (P3HT) and shed light on the complex exciton relaxation dynamics occurring in π-conjugated oligomers of potential interest for optoelectronic applications.


A netting-based alternative to rigid sorting grids in the small-meshed Norway pout (Trisopterus esmarkii) trawl fishery.
A new bycatch reduction device, termed "Excluder", is presented as an alternative to a traditional rigid sorting grid, mandatory in the small-meshed Norway Pout (Trisopterus esmarkii) trawl fishery in the North Sea. The fishery is a high-volume fishery with large vessels, large demersal trawls and catches up to 100 tons per haul of this small forage fish. The Excluder is a 30 m long netting-based sorting system, developed to reduce bycatch (70 mm square meshes) and improving on board gear-handling and safety. The Excluder was tested against a 5.8 m2 standard sorting grid (35 mm bar spacing) in a twin-trawl experiment from the commercial 70 m trawler "S364 Rockall". Catch data were analysed by species and length using the catch comparison method. For all bycatch species analysed, the Excluder had significantly lower catches relative to the grid: herring (21%), whiting (6%), mackerel (5%), American plaice (70%), witch flounder (15%), and lesser silver smelt (71%). For Norway Pout there was a significant increase in the overall catch efficiency of 32%. These results are explained by a 10 cm smaller L50 (the length of fish with 50% probability of being rejected by the sorting system) of the Excluder and a 15 times larger sorting area, which reduces the risk of clogging and loss of function. With these documented effects of improved sorting and target species catch efficiency, implementation of the Excluder would improve sustainability and address two main barriers of the current Norway pout fishery that limit quota capitalization; a tendency for Norway pout to mix with herring and whiting and lowered catch rates from grid-clogging. Additionally, gear-handling and safety on board would be improved.


Removal of the eye-blink artifacts from EEGs via STF-TS modeling and robust minimum variance beamforming.
A novel scheme for the removal of eye-blink (EB) artifacts from electroencephalogram (EEG) signals based on a novel space-time-frequency (STF) model of EEGs and robust minimum variance beamformer (RMVB) is proposed. In this method, in order to remove the artifact, the RMVB is provided with a priori information, namely, an estimation of the steering vector corresponding to the point source EB artifact. The artifact-removed EEGs are subsequently reconstructed by deflation. The a priori knowledge, the vector corresponding to the spatial distribution of the EB factor, is identified using the STF model of EEGs, provided by the parallel factor analysis (PARAFAC) method. In order to reduce the computational complexity present in the estimation of the STF model using the three-way PARAFAC, the time domain is subdivided into a number of segments, and a four-way array is then set to estimate the STF-time/segment (TS) model of the data using the four-way PARAFAC. The correct number of the factors of the STF model is effectively estimated by using a novel core consistency diagnostic- (CORCONDIA-) based measure. Subsequently, the STF-TS model is shown to closely approximate the classic STF model, with significantly lower computational cost. The results confirm that the proposed algorithm effectively identifies and removes the EB artifact from raw EEG measurements.


Salmonella enterica serovar Enteritidis genes induced during oviduct colonization and egg contamination in laying hens.
Salmonella enterica serovar Enteritidis is the predominant serovar associated with salmonellosis worldwide, which is in part due to its ability to contaminate the internal contents of the hen's egg. It has been shown that S. enterica serovar Enteritidis has an unusual tropism for the avian reproductive tract and an ability to persist in the oviduct and ovary. Factors allowing S. enterica serovar Enteritidis strains to contaminate eggs could be a specific interaction with the oviduct tissue, leading to persisting oviduct colonization. In vivo expression technology, a promoter-trap strategy, was used to identify genes expressed during oviduct colonization and egg contamination with S. enterica serovar Enteritidis. A total of 25 clones with in vivo-induced promoters were isolated from the oviduct tissue and from laid eggs. Among the 25 clones, 7 were isolated from both the oviducts and the eggs. DNA sequencing of the cloned promoters revealed that genes involved in amino acid and nucleic acid metabolism, motility, cell wall integrity, and stress responses were highly expressed in the reproductive tract tissues of laying hens.


ART in Europe, 2015: results generated from European registries by ESHRE.
What are the European trends and developments in ART and IUI in 2015 as compared to previous years? The 19th ESHRE report on ART shows a continuing expansion of treatment numbers in Europe, and this increase, the variability in treatment modalities and the rising contribution to the birth rates in most participating countries all point towards the increasing impact of ART on European society. Since 1997, the ART data generated by national registries have been collected, analysed and reported in 18 manuscripts published in Human Reproduction. Collection of European data by the European IVF-Monitoring Consortium (EIM) for ESHRE. The data for treatments performed between 1 January and 31 December 2015 in 38 European countries were provided by national registries or on a voluntary basis by clinics or professional societies. From 1343 institutions in 38 countries offering ART services a total of 849 811 treatment cycles, involving 155 960 with IVF, 385676 with ICSI, 218098 with frozen embryo replacement (FER), 21 041 with preimplantation genetic testing (PGT), 64 477 with egg donation (ED), 265 with IVM and 4294 with FOR were recorded. European data on IUI using husband/partner's semen (IUI-H) and donor semen (IUI-D) were reported from 1352 institutions offering IUI in 25 countries and 21 countries, respectively. A total of 139 050 treatments with IUI-H and 49 001 treatments with IUI-D were included. In 18 countries (14 in 2014) with a population of approximately 286 million inhabitants, in which all institutions contributed to their respective national registers, a total of 409 771 treatment cycles were performed, corresponding to 1432 cycles per million inhabitants (range: 727-3068 per million). After IVF the clinical pregnancy rates (PRs) per aspiration and per transfer were slightly lower in 2015 as compared to 2014, at 28.5 and 34.6% versus 29.9 and 35.8%, respectively. After ICSI, the corresponding PR achieved per aspiration and per transfer in 2015 were also slightly lower than those achieved in 2014 (26.2 and 33.2% versus 28.4 and 35.0%, respectively). On the other hand, after FER with own embryos the PR per thawing continued to rise from 27.6% in 2014 to 29.2% in 2015. After ED a slightly lower PR per embryo transfer was achieved: 49.6% per fresh transfer (50.3% in 2014) and 43.4% for FOR (48.7% in 2014). The delivery rates (DRs) after IUI remained stable at 7.8% after IUI-H (8.5% in 2014) and at 12.0% after IUI-D (11.6% in 2014). In IVF and ICSI together, 1, 2, 3 and ≥4 embryos were transferred in 37.7, 53.9, 7.9 and in 0.5% of all treatments, respectively (corresponding to 34.9, 54.5, 9.9 and in 0.7% in 2014). This evolution towards the transfer of fewer embryos in both IVF and ICSI resulted in a proportion of singleton, twin and triplet DR of 83.1, 16.5 and 0.4%, respectively (compared to 82.5, 17.0 and 0.5%, respectively, in 2014). Treatments with FER in 2015 resulted in twin and triplet DR of 12.3 and 0.3%, respectively (versus 12.4 and 0.3% in 2014). Twin and triplet delivery rates after IUI-H were 8.9 and 0.5%, respectively (in 2014: 9.5 and 0.3%), and 7.3 and 0.6% after IUI-D (in 2014: 7.7 and 0.3%). The methods of data collection and reporting vary among European countries. The EIM receives aggregated data from various countries with variable levels of completeness. Registries from a number of countries have failed to provide adequate data about the number of initiated cycles and deliveries. As long as incomplete data are provided, the results should be interpreted with caution. The 19th EIM report on ART shows a continuing expansion of treatment numbers in Europe. The number of treatments reported, the variability in treatment modalities and the rising contribution to the birth rates in most participating countries point towards the increasing impact of ART on reproduction in Europe. Being the largest data collection on ART worldwide, detailed information about ongoing developments in the field is provided. The study has no external funding and all costs are covered by ESHRE. There are no competing interests.


Low-Intensity and Short-Duration Continuous Cervical Transcutaneous Spinal Cord Stimulation Intervention Does Not Prime the Corticospinal and Spinal Reflex Pathways in Able-Bodied Subjects.
Cervical transcutaneous spinal cord stimulation (tSCS) has been utilized in applications for improving upper-limb sensory and motor function in patients with spinal cord injury. Although therapeutic effects of continuous cervical tSCS interventions have been reported, neurophysiological mechanisms remain largely unexplored. Specifically, it is not clear whether sub-threshold intensity and 10-min duration continuous cervical tSCS intervention can affect the central nervous system excitability. Therefore, the purpose of this study was to investigate effects of sub-motor-threshold 10-min continuous cervical tSCS applied at rest on the corticospinal and spinal reflex circuit in ten able-bodied individuals. Neurophysiological assessments were conducted to investigate (1) corticospinal excitability via transcranial magnetic stimulation applied on the primary motor cortex to evoke motor-evoked potentials (MEPs) and (2) spinal reflex excitability via single-pulse tSCS applied at the cervical level to evoke posterior root muscle (PRM) reflexes. Measurements were recorded from multiple upper-limb muscles before, during, and after the intervention. Our results showed that low-intensity and short-duration continuous cervical tSCS intervention applied at rest did not significantly affect corticospinal and spinal reflex excitability. The stimulation duration and/or intensity, as well as other stimulating parameters selection, may therefore be critical for inducing neuromodulatory effects during cervical tSCS.


Methylation status of Wnt signaling pathway genes affects the clinical outcome of Philadelphia-positive acute lymphoblastic leukemia.
The clinical significance of aberrant promoter methylation of the canonical Wnt pathway antagonist genes (sFRP1, sFRP2, sFRP4, sFRP5, Wif1, Dkk3, and Hdpr1) and also putative tumor-suppressor gene Wnt5a, belonging to the non-canonical Wnt signaling pathway, was investigated in a large series of 75 patients with Philadelphia chromosome-positive acute lymphoblastic leukemia by methylation-specific polymerase chain reaction. At least one methylated gene was observed in cells from 66% (49/75) of patients (methylated group). Disease-free survival and overall survival at 9 years were 51 and 40%, respectively, for the unmethylated group and 3 and 2%, respectively, for the methylated group (both P < 0.0001). Multivariate analysis demonstrated that the Wnt methylation profile was an independent prognostic factor predicting disease-free survival (P = 0.007) and overall survival (P = 0.039). Abnormal DNA methylation of promoter-associated CpG islands in the Wnt signaling pathway is very common in Philadelphia chromosome-positive acute lymphoblastic leukemia and potentially defines subgroups with distinct clinical characteristics.


A 30-month-old child with acute renal failure due to primary renal cytotoxic T-cell lymphoma.
We present a case of a 30-month-old child who presented with anemia and acute renal failure, and was found to have bilateral renal involvement by primary cytotoxic T-cell lymphoma. This was characterized by a monotonous interstitial lymphoid infiltrate with extensive necrosis. The tumor cells showed a CD8, granzyme, and TIA1-positive phenotype with no evidence of Epstein-Barr virus by in situ hybridization. The differential diagnosis based on the biopsy findings included a reactive interstitial nephritis; however, molecular studies confirmed T-cell clonality. She was started on induction chemotherapy and subsequently received maintenance therapy with methotrexate and 6-mercaptopurine. The patient had a complete response after chemotherapy and at 21 months of follow-up, she has no evidence of residual lymphoma; however, she has developed a dilated cardiomyopathy and she remains in renal failure. We discuss the morphologic, immunophenotypic, and molecular features of our case and describe the clinical course of our patient. We review the literature on primary renal lymphoma with an emphasis on T-lineage lymphomas and those that occur in children.


Blood-group-related carbohydrate antigens are expressed on human milk galactosyltransferase and are immunogenic in rabbits.
Immunochemical evidence is presented for the presence of blood-group-related carbohydrate structures on human milk galactosyltransferase and for the occurrence of the corresponding specificities among rabbit antibodies to this enzyme. Although these carbohydrate specificities constitute minor populations among antisera and affinity-purified antibodies to galactosyltransferase, their presence is important in the immunohistochemical approach to enzyme localization, since they give rise to strong reactivities with epithelial cells of the gastrointestinal tract.


PEGylated degradable composite nanoparticles based on mixtures of PEG-b-poly(γ-benzyl L-glutamate) and poly(γ-benzyl L-glutamate).
In the present work, the possibility to obtain PEGylated nanoparticles from two PBLG derivatives, PEG-b-poly(γ-benzyl L-glutamate), PBLG-PEG-60, and poly(γ-benzyl L-glutamate), PBLG-Bnz-50, by nanoprecipitation has been investigated. Particles were prepared not only from one polymer (PBLG-PEG-60 or PBLG-Bnz-50), but also from mixtures of two PBLG derivatives, PBLG-PEG-60 and PBLG-Bnz-50, in different ratios (90/10, 77/23, and 40/60 wt %). Because of the presence of PEG chains, hydrophilic particles were obtained, which was confirmed by ζ potential measurements (ζ from -13 mV and -21 mV) and by isothermal titration microcalorimetry (ITC). This last technique has shown no heat exchange when BSA was added to PEGylated nanoparticles. Further, complement activation has been evaluated by crossed immuno-electrophoresis demonstrating that the introduction of 77 wt % of PEGylated PBLG chains in the particles was enough to ensure a low complement activation activity. This effect was strongly correlated to the ζ potential of the particles, which decreased with an increase of PEG chains content. Interestingly, such properties are of interest for the preparation of degradable stealth nanocarriers. Moreover, it is suggested that the introduction of a reasonable amount (up to 20 wt %) of a second copolymer in the particle composition can be possible without modifying their stealth character. Moreover, the presence of this second copolymer would help to introduce a second functionality to the particles.


Living in an adaptive world: Genomic dissection of the genus Homo and its immune response.
More than a decade after the sequencing of the human genome, a deluge of genome-wide population data are generating a portrait of human genetic diversity at an unprecedented level of resolution. Genomic studies have provided new insight into the demographic and adaptive history of our species, Homo sapiens, including its interbreeding with other hominins, such as Neanderthals, and the ways in which natural selection, in its various guises, has shaped genome diversity. These studies, combined with functional genomic approaches, such as the mapping of expression quantitative trait loci, have helped to identify genes, functions, and mechanisms of prime importance for host survival and involved in phenotypic variation and differences in disease risk. This review summarizes new findings in this rapidly developing field, focusing on the human immune response. We discuss the importance of defining the genetic and evolutionary determinants driving immune response variation, and highlight the added value of population genomic approaches in settings relevant to immunity and infection.


Determinants of incidence and clearance of high-risk human papillomavirus infections in rural Rakai, Uganda.
We used self-administered vaginal swabs to assess the incidence and clearance of carcinogenic human papillomavirus (HPV) infections in rural Rakai, Uganda. Women provided self-administered vaginal swab at annual home-based visits. Type-specific carcinogenic HPV incidence and clearance and risk factors were assessed. Carcinogenic HPV incidence was 17.3 per 100 person-years among HIV-positive women compared with 7.0 per 100 person-years among HIV-negative women (P < 0.001). HPV-51 had the highest incidence followed by HPV-16 (1.8 per 100 and 1.5 per 100 person-years, respectively). In multivariate model, HIV-positive women were twice as likely to have incident infection compared with HIV-negative women. Younger women were at higher risk for incident infection, as were women with higher lifetime and recent sexual partners, and high perception of AIDS. Married women were less likely to have incident infection. Approximately half of all carcinogenic HPV infections cleared over the study follow-up of 3 years. HPV-31, HPV-35, and HPV-16 had the lowest clearance (16.7%, 27.9%, and 38.3%, respectively). In multivariate model, HIV-positive, women over 30 years with higher HPV viral, burden and more lifetime sex partners were less likely to clear infections. Self-collected vaginal swabs provide accurate HPV exposure assessment for studying HPV exposure and epidemiology and can be an important tool for research in populations unwilling to undergo pelvic exam.


Comparison of ximelagatran with warfarin for the prevention of venous thromboembolism after total knee replacement.
In a previous study of the prevention of venous thromboembolism after total knee replacement, the efficacy of ximelagatran, an oral direct thrombin inhibitor that does not require monitoring of coagulation or dose adjustment, was found to be similar to that of warfarin at a dose of 24 mg of ximelagatran twice daily. The purpose of the present study was to determine whether a higher dose of ximelagatran is superior to warfarin. This randomized, double-blind trial compared a regimen of 7 to 12 days of oral ximelagatran, at a dose of 24 or 36 mg twice daily, starting the morning after surgery, with warfarin therapy started the evening of the day of surgery. The composite end point of venous thromboembolism and death from all causes and the incidence of bleeding were the primary outcome measures. Among the 1851 patients in the efficacy analysis, oral ximelagatran at a dose of 36 mg twice daily was superior to warfarin with respect to the primary composite end point of venous thromboembolism and death from all causes (20.3 percent vs. 27.6 percent; P=0.003). There were no significant differences between these two groups with respect to major bleeding (incidence, 0.8 percent and 0.7 percent, respectively), perioperative indicators of bleeding, wound characteristics, or the composite secondary end point of proximal deep-vein thrombosis, pulmonary embolism, and death (2.7 percent vs. 4.1 percent; P=0.17). The efficacy of oral ximelagatran, administered starting the morning after total knee replacement, was superior to that of warfarin for prevention of venous thromboembolism. Rates of hemorrhagic complications with the two drugs were similar.


Experimental facial transplantation surgery.
Facial composite tissue allograft transplantation represents a novel frontier in the reconstruction of complex human facial defects. To develop more refined surgical techniques and yield fine results, it is definite to make a suitable animal model. Rabbit is a medium-sized animal and considered to have many advantages than any other animal, but published reports are very few and contain less detailed description. We made an easy and accessible facial transplantation approach and tried to ascertain its virtue of use as an appropriate transplantation model. The facial-scalp flap including ipsilateral ear was harvested and transferred as a single unit with unilateral common carotid artery and external jugular vein. Most allografts survived with perfect viability without vascular problems in the early postoperative period, and rejection response was also identified by clinical and histopathologic analysis. The total mean time of transplantation was 2 hours +/- 10 minutes. We documented that this model is well qualified for use as a standard transplantation training model and future research work in every aspect.


A critical review of protocols for moss biomonitoring of atmospheric deposition: sampling and sample preparation.
Currently, the most important guideline for the application of the moss technique to monitor the atmospheric deposition of heavy metals is the "Heavy metals, nitrogen and POPs in European mosses: 2015 survey" published by the UNECE ICP Vegetation. Two main problems have been identified with this guideline: i) some of the recommendations regarding the methodological aspects involved in the application of the moss technique are not based on scientific criteria; and, ii) some recommendations in the manual are very vague and some aspects are even left out (e.g., elevation, distance to the coast). As a result there exists a high variability in the application of the protocol and many scientists adapt it to the specific conditions in the studied areas without evaluating how changes affect the results obtained. Therefore, in this article a total of 369 studies were reviewed including both methodological and application studies of the passive biomonitoring of the atmospheric deposition of heavy metals with terrestrial mosses. The results of this review have shown on the one hand, that none of the articles completely accomplished the ICP-Vegetation protocol suggestions, either because the information regarding some aspects was lacking or simply because the authors did not follow the manual suggestions. On the other hand, it was found that the results of methodological studies sometimes contradicted the ICP Vegetation manual recommendations. Thus, a new protocol in which each suggestion has been carefully and rigorously contrasted with the available literature has been proposed in this paper. In addition, practical and economic issues have also been considered and much more concise suggestions have been proposed which would facilitate its fulfilment in a more objective way.


Prevalence of Sarcocystis spp. in two subspecies of caribou (Rangifer tarandus) in Newfoundland and Labrador, and foxes (Vulpes vulpes), wolves (Canis lupus), and husky dogs (Canis familiaris) as potential definitive hosts.
A study was conducted to determine the prevalence and geographical distribution of Sarcocystis spp. infecting 2 subspecies of caribou (Rangifer tarandus) inhabiting Newfoundland and Labrador and its potential definitive hosts. Muscle samples of caribou were obtained, primarily from hunters, and feces of red foxes (Vulpes vulpes) and wolves (Canis lupus), from trappers, and Husky dogs (Canis familiaris), from owners. Histological sections of muscle and flotation methods for feces were used for parasitic detection. Sarcocystis sp. infected more than 50% of barren-ground caribou (R. t. tarandus) from 4 locations in Newfoundland, but it was significantly greater in the north, where 99% of woodland caribou (R. t. caribou) from Labrador harbored the infection. Sporocysts were observed in 27 of 32 red foxes from eastern and northern Newfoundland, whereas 15 of 15 wolves and 22 of the 38 Husky dogs were infected. Wolves and red foxes probably acquired the infection through scavenging, and Husky dogs, from meat they were fed.


Biosatellite 3: a physiological interpretation.
On 28 June 1969, a macaque monkey was boosted into orbit for a projected 30-day flight. It was aborted after 8 1/2 days and the monkey died 16 hours later. Data collected indicate: Weightlessness and the hypothermia acted to shift blood volume centrally; this provided a strong drive for the reduction of blood volume. Restraint, unusual vestibular sensations, and the continuing polydipsia all acted to disturb the central mechanisms affecting salt and water metabolism; it is probable that the function of the kidney was significantly affected and that an excessive amount of salt was lost. We can speculate that a serious electrolyte disturbance was superimposed on growing dehydration. Unpleasant vestibular sensations may have contributed to the high evaporative loss by autonomic disturbance, the whole problem being compounded and reinforced by the unnatural restraint to which the monkey was subjected.


Foreword.
"Mini-Reviews in Medicinal Chemistry" is now commencing its 14th year. It has grown over the years to a reputable international journal known for rapidly publishing exciting new researches in the field. The journal owes its success and popularity to the very hard work of the Co-Editors, Associate Editors, Regional Editors and Editorial Advisory Board Members to whom I am grateful for their kind guidance and enthusiastic support. I am also very grateful to the hard working staff of Bentham Science Publishers including Ms. Sara Arif, Mrs. Mehwish Akhter and Mr. Muzaffar Hussain. I look forward to another good year for the journal with the publication of high quality publications from eminent experts"


Prognostic Factors of Survival with Aflibercept and FOLFIRI (fluorouracil, leucovorin, irinotecan) as Second-line Therapy for Patients with Metastatic Colorectal Cancer.
Background: Aflibercept and fluorouracil, leucovorin, irinotecan (FOLFIRI) is commonly used as a second-line treatment for metastatic colorectal cancer (CRC). However, the biomarkers to guide the choice of this regimen from among treatment options remain unclear. Patients and Methods: We performed exploratory analyses to validate potential prognostic factors for patients receiving aflibercept plus FOLFIRI as a second-line systemic treatment for metastatic CRC between January 2015 and July 2019. Patient characteristics, histopathologic data, laboratory and radiologic data, and treatment outcomes were collected and reviewed. Results: Included were 52 patients: 50 (96.2%) received bevacizumab plus fluorouracil, leucovorin, oxaliplatin (FOLFOX) as prior first-line treatment. Among the 52 patients receiving aflibercept and FOLFIRI, four complete responses and 21 partial responses were observed in analyzed patients for an overall response rate of 48.1%. Median progression-free survival (PFS) was 7.0 months and overall survival (OS) was 16.8 months. Response to first-line treatment (median PFS, 8.0 versus 4.2 months), left-side location of primary tumor (7.9 versus 4.9 months), low baseline CEA level (8.0 versus 5.9 months), and no RAS/RAF mutation (9.9 versus 6.4 months) were remained significant prognostic factors for PFS in the multivariate backward stepwise Cox regression model, and the latter three factors were also significantly related to OS. Conclusions: Significant prognostic factors for PFS with aflibercept plus FOLFIRI as second-line therapy were extracted and validated in the multivariate OS model. These findings could provide useful information for selecting patients for aflibercept plus FOLFIRI as second-line therapy.


Polymerase chain reaction detection of avipox and avian papillomavirus in naturally infected wild birds: comparisons of blood, swab and tissue samples.
Avian poxvirus (avipox) is widely reported from avian species, causing cutaneous or mucosal lesions. Mortality rates of up to 100% are recorded in some hosts. Three major avipox clades are recognized. Several diagnostic techniques have been reported, with molecular techniques used only recently. Avipox has been reported from 278 different avian species, but only 111 of these involved sequence and/or strain identification. Collecting samples from wild birds is challenging as only few wild bird individuals or species may be symptomatic. Also, sampling regimes are tightly regulated and the most efficient sampling method, whole bird collection, is ethically challenging. In this study, three alternative sampling techniques (blood, cutaneous swabs and tissue biopsies) from symptomatic wild birds were examined. Polymerase chain reaction was used to detect avipoxvirus and avian papillomavirus (which also induces cutaneous lesions in birds). Four out of 14 tissue samples were positive but all 29 blood samples and 22 swab samples were negative for papillomavirus. All 29 blood samples were negative but 6/22 swabs and 9/14 tissue samples were avipox-positive. The difference between the numbers of positives generated from tissue samples and from swabs was not significant. The difference in the avipox-positive specimens in paired swab (4/6) and tissue samples (6/6) was also not significant. These results therefore do not show the superiority of swab or tissue samples over each other. However, both swab (6/22) and tissue (8/9) samples yielded significantly more avipox-positive cases than blood samples, which are therefore not recommended for sampling these viruses.


The use of nanocrystalline cellulose for the binding and controlled release of drugs.
The objective of this work was to investigate the use of nanocrystalline cellulose (NCC) as a drug delivery excipient. NCC crystallites, prepared by an acid hydrolysis method, were shown to have nanoscopic dimensions and exhibit a high degree of crystallinity. These crystallites bound significant quantities of the water soluble, ionizable drugs tetratcycline and doxorubicin, which were released rapidly over a 1-day period. Cetyl trimethylammonium bromide (CTAB) was bound to the surface of NCC and increased the zeta potential in a concentration-dependent manner from -55 to 0 mV. NCC crystallites with CTAB-modified surfaces bound significant quantities of the hydrophobic anticancer drugs docetaxel, paclitaxel, and etoposide. These drugs were released in a controlled manner over a 2-day period. The NCC-CTAB complexes were found to bind to KU-7 cells, and evidence of cellular uptake was observed.


Epigenetic regulation of facultative heterochromatinisation in Planococcus citri via the Me(3)K9H3-HP1-Me(3)K20H4 pathway.
Using RNA interference (RNAi) we have conducted a functional analysis of the HP1-like chromobox gene pchet2 during embryogenesis of the mealybug Planococcus citri. Knocking down pchet2 expression results in decondensation of the male-specific chromocenter that normally arises from the developmentally-regulated facultative heterochromatinisation of the paternal chromosome complement. Together with the disappearance of the chromocenter the staining levels of two associated histone modifications, tri-methylated lysine 9 of histone H3 [Me(3)K9H3] and tri-methylated lysine 20 of histone H4 [Me(3)K20H4], are reduced to undetectable levels. Embryos treated with double-stranded RNA (dsRNA) targeting pchet2 also exhibit chromosome abnormalities, such as aberrant chromosome condensation, and also the presence of metaphases that contain 'lagging' chromosomes. We conclude that PCHET2 regulates chromosome behavior during metaphase and is a crucial component of a Me(3)K9H3-HP1-Me(3)K20H4 pathway involved in the facultative heterochromatinisation of the (imprinted) paternal chromosome set.


Shift workers have a similar diet quality but higher energy intake than day workers.
Shift work is associated with adverse health outcomes, and an unhealthy diet may be a contributing factor. We compared diet quantity and quality between day and shift workers, and studied exposure-response relationships regarding frequency of night shifts and years of shift work. Cross-sectional general population data from the European Prospective Investigation into Cancer and Nutrition-Netherlands (EPIC-NL) cohort was used. Dietary intake was assessed in 1993-1997 among adults aged 20-70 years using a food frequency questionnaire. We calculated energy intake, the Mediterranean Diet Score (MDS) and WHO-based Healthy Diet Indicator (HDI). In 2011-2014, we retrospectively identified 683 shift workers and 7173 day workers in 1993-1997. Using multivariable-adjusted linear regression analysis, we estimated regression coefficients (β) and 95% confidence intervals (95% CI) of the differences in dietary intake between day and shift workers. Shift workers had a higher energy intake than day workers (β:56 kcal/d, 95% CI 10-101), and a higher consumption of grains, dairy products, meat and fish (P<0.05). The difference in energy intake was largest for shift workers with ≥5 night shifts/month. They consumed 103 kcal/d (95% CI 29-176) more than day workers. No associations were found with MDS and HDI. Shift workers and particularly those with a high frequency of night shifts had a higher energy intake than day workers. Regardless of number of night shifts and years of shift work, shift workers had similar diet quality as day workers. This suggests that increased energy intake among shift workers may contribute to shift work-induced adverse health outcomes.


Asthma and major depressive episode in adolescents in France.
The association between asthma and anxiety disorders in teenagers is well documented, but data about the association with mood disorders are scarce. We analyzed data from a cross-sectional study conducted among ninth grade schoolchildren in France in 2003-2004. The teenagers were selected by two-stage sampling and interviewed by school doctors/nurses using a standardized questionnaire including questions about asthma and asthma-like symptoms. They also completed a self-administered questionnaire in which the occurrence of major depressive episodes (MDEs) during the past 12 months was assessed by the Composite International Diagnostic Interview-Short Form. A total of 7000 teenagers (mean age 15.1 years) were included. The prevalence of wheezing in the past 12 months was 10.0% and that of current asthma (wheezing in the past 12 months in children who had already had asthma attacks, or treatment for wheezing or asthma in the past 12 months) was 8.5%. The prevalence of MDE during the past year was 14.2% in teenagers with current asthma versus 9.2% among the others. The association between current asthma and past-year MDE remained significant after adjustment for age, gender, family structure, and the father's employment status. Asthma was uncontrolled (at least four attacks of wheezing, one awakening per week due to wheezing, one severe wheezing, four unplanned medical visits, or one hospitalization for a wheezing attack in the past year) in more than half (58.3%) of asthmatic teenagers with an MDE in the past year versus 35.3% of those without an MDE. Asthma is associated with a higher prevalence of MDE. Among adolescents with asthma, MDE is associated with poorer asthma control. These findings highlight the need for a comprehensive care management of asthma in France that takes the psychological dimension into account.


A mouse model of familial hypertrophic cardiomyopathy caused by a alpha-tropomyosin mutation.
Familial hypertrophic cardiomyopathy, a disease caused by mutations in cardiac contractile proteins, is characterized by left and/or right ventricular hypertrophy, myocyte disarray, fibrosis, and cardiac arrhythmias that may lead to premature sudden death. Five distinct point mutations within alpha-tropomyosin are associated with the development of familial hypertrophic cardiomyopathy. Two of these mutations are found within a troponin T binding site, located at amino acids 175 and 180. In this study, we analyze a transgenic mouse model for one of the mutations that occur at codon 180: a substitution of a glutamic acid for a glycine. These mice develop severe cardiac hypertrophy, substantial interstitial fibrosis, and have an increased heart weight/ body weight ratio. Results show that calcium-handling proteins associated with the sarcoplasmic reticulum exhibit decreased expression. These alterations in gene expression, coupled with the structurally-altered tropomyosin, may contribute to the demonstrated decreased physiological performance exhibited by these transgenic mice. A DNA hybridization microarray analysis of the transgenic vs. control ventricular RNAs shows that 50 transcripts are differentially expressed by more than 100% during the onset of the hypertrophic process, many of which are associated with the extracellular matrix. This study demonstrates that mutations within tropomyosin can be severely disruptive of sarcomeric function, triggering a hypertrophic response coupled with a cascade of alterations in gene expression.


Preferential glutathione conjugation of a reverse diol epoxide compared to a bay region diol epoxide of phenanthrene in human hepatocytes: relevance to molecular epidemiology studies of glutathione-s-transferase polymorphisms and cancer.
Bay region diol epoxides are recognized ultimate carcinogens of polycyclic aromatic hydrocarbons (PAH), and in vitro studies have demonstrated that they can be detoxified by conjugation with glutathione, leading to the widely investigated hypothesis that individuals with low activity forms of glutathione-S-transferases are at higher risk of PAH induced cancer, a hypothesis that has found at most weak support in molecular epidemiology studies. A weakness in this hypothesis was that the mercapturic acids resulting from the conjugation of PAH bay region diol epoxides had never been identified in human urine. We recently analyzed smokers' urine for mercapturic acids derived from phenanthrene, the simplest PAH with a bay region. The only phenanthrene diol epoxide-derived mercapturic acid in smokers' urine was produced from the reverse diol epoxide, anti-phenanthrene-3,4-diol-1,2-epoxide (11), not the bay region diol epoxide, anti-phenanthrene-1,2-diol-3,4-epoxide (10), which does not support the hypothesis noted above. In this study, we extended these results by examining the conjugation of phenanthrene metabolites with glutathione in human hepatocytes. We identified the mercapturic acid N-acetyl-S-(r-4,t-2,3-trihydroxy-1,2,3,4-tetrahydro-c-1-phenanthryl)-L-cysteine (14a), (0.33-35.9 pmol/mL at 10 microM 8, 24 h incubation, N = 10) in all incubations with phenanthrene-3,4-diol (8) and the corresponding diol epoxide 11, but no mercapturic acids were detected in incubations with phenanthrene-1,2-diol (7), and only trace amounts were observed in incubations with the corresponding bay region diol epoxide 10. Taken together with our previous results, these studies clearly demonstrate that glutathione conjugation of a reverse diol epoxide of phenanthrene is favored over conjugation of a bay region diol epoxide. Since reverse diol epoxides of PAH are generally weakly or nonmutagenic/carcinogenic, these results, if generalizable to other PAH, do not support the widely held assumption that glutathione-S-transferases are important in the detoxification of PAH in humans.


Topographic analysis of eyelid position using digital image processing software.
To propose a novel analysis technique for objective quantification of topographic eyelid position with an algorithmatically calculated scheme and to determine its feasibility. One hundred normal eyelids from 100 patients were segmented using a graph cut algorithm, and 11 shape features of eyelids were semi-automatically quantified using in-house software. To evaluate the intra- and inter-examiner reliability of this software, intra-class correlation coefficients (ICCs) were used. To evaluate the diagnostic value of this scheme, the correlations between semi-automatic and manual measurements of margin reflex distance 1 (MRD1) and margin reflex distance 2 (MRD2) were analysed using a Bland-Altman analysis. To determine the degree of agreement according to manual MRD length, the relationship between the variance of semi-automatic measurements and the manual measurements was evaluated using linear regression. Intra- and inter-examiner reliability were excellent, with ICCs ranging from 0.913 to 0.980 in 11 shape features including MRD1, MRD2, palpebral fissure, lid perimeter, upper and lower lid lengths, roundness, total area, and medial, central, and lateral areas. The correlations between semi-automatic and manual MRDs were also excellent, with better correlation in MRD1 than in MRD2 (R = 0.893 and 0.823, respectively). In addition, significant positive relationships were observed between the variance and the length of MRD1 and 2; the longer the MRD length, the more the variance. The proposed novel optimized integrative scheme, which is shown to have high repeatability and reproducibility, is useful for topographic analysis of eyelid position.


Genetic determinants of dabigatran plasma levels and their relation to bleeding.
Fixed-dose unmonitored treatment with dabigatran etexilate is effective and has a favorable safety profile in the prevention of stroke in atrial fibrillation patients compared with warfarin. We hypothesized that genetic variants could contribute to interindividual variability in blood concentrations of the active metabolite of dabigatran etexilate and influence the safety and efficacy of dabigatran. We successfully conducted a genome-wide association study in 2944 Randomized Evaluation of Long-term Anticoagulation Therapy (RE-LY) participants. The CES1 single-nucleotide polymorphism rs2244613 was associated with trough concentrations, and the ABCB1 single-nucleotide polymorphism rs4148738 and the CES1 single-nucleotide polymorphism rs8192935 were associated with peak concentrations at genome-wide significance (P<9×10(-8)) with a gene-dose effect. Each minor allele of the CES1 single-nucleotide polymorphism rs2244613 was associated with lower trough concentrations (15% decrease per allele; 95% confidence interval, 10-19; P=1.2×10(-8)) and a lower risk of any bleeding (odds ratio, 0.67; 95% confidence interval, 0.55-0.82; P=7×10(-5)) in dabigatran-treated participants, with a consistent but nonsignificant lower risk of major bleeding (odds ratio, 0.66; 95% confidence interval, 0.43-1.01). The interaction between treatment (warfarin versus all dabigatran) and carrier status was statistically significant (P=0.002), with carriers having less bleeding with dabigatran than warfarin (hazard ratio, 0.59; 95% confidence interval, 0.46-0.76; P=5.2×10(-)5) in contrast to no difference in noncarriers (hazard ratio, 0.96; 95% confidence interval, 0.81-1.14; P=0.65). There was no association with ischemic events, and neither rs4148738 nor rs8192935 was associated with bleeding or ischemic events. Genome-wide association analysis identified that carriage of the CES1 rs2244613 minor allele occurred in 32.8% of patients in RE-LY and was associated with lower exposure to active dabigatran metabolite. The presence of the polymorphism was associated with a lower risk of bleeding. URL: http://www.clinicaltrials.gov. Unique identifier: NCT00262600.


Mutations in the substrate binding site of human heat-shock protein 70 indicate specific interaction with HLA-DR outside the peptide binding groove.
Heat-shock protein 70 (Hsp70)-peptide complexes are involved in MHC class I- and II-restricted antigen presentation, enabling enhanced activation of T cells. As shown previously, mammalian cytosolic Hsp70 (Hsc70) molecules interact specifically with HLA-DR molecules. This interaction might be of significance as Hsp70 molecules could transfer bound antigenic peptides in a ternary complex into the binding groove of HLA-DR molecules. The present study provides new insights into the distinct interaction of Hsp70 with HLA-DR molecules. Using a quantitative binding assay, it could be demonstrated that a point mutation of amino acids alanine 406 and valine 438 in the substrate binding pocket led to reduced peptide binding compared with the wild-type Hsp70 whereas HLA-DR binding remains unaffected. The removal of the C-terminal lid neither altered the substrate binding capacity nor the Hsp70 binding characteristics to HLA-DR. A truncated variant lacking the nucleotide binding domain showed no binding interactions with HLA-DR. Furthermore, the truncated ATPase subunit of constitutively expressed Hsc70 revealed similar binding affinities to HLA-DR compared with the complete Hsc70. Hence, it can be assumed that the Hsp70-HLA-DR interaction takes place outside the peptide binding groove and is attributed to the ATPase domain of HSP70 molecules. The Hsp70-chaperoned peptides might thereby be directly transferred into the binding groove of HLA-DR, so enabling enhanced presentation of the peptide on antigen-presenting cells and leading to an improved proliferation of responding T cells as shown previously.


Enrofloxacin-induced retinopathy in a guanaco (Lama guanicoe).
A 4-yr-old male guanaco (Lama guanicoe) in a multispecies exhibit presented with a laceration in the axillary region. The laceration was surgically repaired. Medical treatment was initiated with penicillin G procaine and benzathine (1920 IU/kg, i.m., s.i.d. for 14 days), and enrofloxacin (2.4 mg/kg, i.m., s.i.d. for 14 days). The animal was later treated with trimethoprim-sulfadiazine (24 mg/kg, p.o., s.i.d. for 10 days). Twenty-six days after initial presentation, the guanaco was suspected to be blind. An ophthalmic examination confirmed retinal damage. On postmortem histological evaluation, there was outer retinal atrophy that was most severe in the central retina with localized foci of complete retinal atrophy; lesions were similar to those observed in enrofloxacin retinal toxicity in cats.


Transposition of Tn4551 in Bacteroides fragilis: identification and properties of a new transposon from Bacteroides spp.
Tn4551, a clindamycin resistance (Ccr) transposon from the R plasmid pBI136, was cloned onto an Escherichia coli-Bacteroides shuttle vector which could replicate normally in E. coli but was maintained unstably in Bacteroides fragilis. To aid in cloning and to ensure maintenance of Tn4551 in E. coli, a kanamycin resistance determinant (Kmr) was inserted in the transposon. The transposon-bearing shuttle vector pFD197 was transformed into B. fragilis 638, and putative insertions of Tn4551::Kmr were identified by screening for resistance to clindamycin and plasmid content. Southern hybridization analyses were used to verify integration of the transposon in the B. fragilis chromosome, and the frequency of insertion was estimated at 7.8 X 10(-5) events per generation. In 57% of the isolates tested a second integration event also occurred. This second insertion apparently involved just a single copy of the 1.2-kilobase repeat sequence which flanks the transposon. In addition, Tn4551::Kmr appeared to function as a transposon in E. coli. Evidence for this was obtained by the isolation of transposon insertions into the bacteriophage P1 genome. Finally, the transposon vector, pFD197, could be mobilized to other B. fragilis strains in which transposition was detected. Mobilization from the strain 638 background was via a conjugation like process, but occurred in the absence of known conjugative elements or other detectable plasmids. This result suggested the presence of a host-encoded transfer system in this B. fragilis strain.


Esophageal symptoms questionnaire for the assessment of dysphagia, globus, and reflux symptoms: initial development and validation.
Esophageal symptoms often co-occur. A validated self-report measure encompassing multiple esophageal symptoms is necessary to determine their frequency and severity both independently and in association with each other. Such a questionnaire could streamline the diagnostic process and guide patient management. We aimed to develop an integrative measure that provides a clinical 'snapshot' of common esophageal symptoms. Internal reliability and content validity of a 38-item self-report Esophageal Symptoms Questionnaire (ESQ), measuring the frequency and severity of typical esophageal symptoms using Likert-rating scales were assessed in 211 patients presenting to gastroenterology and ENT outpatient tertiary care clinics. Reproducibility, concurrent and predictive validity were evaluated using the reduced-item ESQ. The 38-item ESQ had high internal reliability. Principal component analyses and item reduction methods identified three components, to which 30 of 38 items contributed significantly, providing 59% of total variance. The test-retest correlations were moderate-to-strong for 24 of 30 new items (r(s) ≥ 0.44, P < 0.05). The resultant subscales measuring dysphagia (ESQ-D), globus (ESQ-G), and reflux (ESQ-R) compared well against concurrent physician's 'working' diagnosis (odds ratio 1.04-1.09). The receiver operating characteristics were adequate-to-good for ESQ-D (area under the curve [AUC]= 0.87) and ESQ-G (AUC = 0.74), but poor for ESQ-R (AUC = 0.61) although it matched the content of the validated Reflux Disease Questionnaire. The brief 30-item ESQ shows good internal reliability and content validity as a summary of the extent of dysphagia, globus and reflux symptoms. As a tool measuring more than one esophageal symptom, ESQ could guide patient management by indicating which of the coexisting symptoms needs to be addressed first.


[Iris-corneal dysgenesis].
This work presents a family, a mother and her 3 daughters, aged: 10 months, 4 years, 5 years with Rieger's anomaly. For children, this anomaly is associated with glaucoma and buphtalmy, this last element is the one that made the family ask for medical assistance and come to the hospital. The study of the disease was done on five generations in the same family, on the mother's line. This revealed that in the mother's family the grand grandmother and her two sisters were blind at an early age (30-40 years old). The presence of the anomaly in many generations proves the genetic transmission of the disease. We have to mention that in the last generation, the most serious appearance was registered at the patient with the earliest age (10 months).


A randomized trial of vitamin D₃ supplementation in children: dose-response effects on vitamin D metabolites and calcium absorption.
Changes in serum vitamin D metabolites and calcium absorption with varying doses of oral vitamin D₃ in healthy children are unknown. Our objective was to examine the dose-response effects of supplemental vitamin D₃ on serum vitamin D metabolites and calcium absorption in children living at two U.S. latitudes. Black and white children (n = 323) participated in a multisite (U.S. latitudes 34° N and 40° N), triple-masked trial. Children were randomized to receive oral vitamin D₃ (0, 400, 1000, 2000, and 4000 IU/d) and were sampled over 12 weeks in winter. Serum 25-hydroxyvitamin D (25(OH)D) and 1,25-dihydroxyvitamin D (1,25(OH)₂D) were measured using RIA and intact PTH (iPTH) by immunoradiometric assay. Fractional calcium absorption was determined from an oral stable isotope ⁴⁴Ca (5 mg) in a 150-mg calcium meal. Nonlinear and linear regression models were fit for vitamin D metabolites, iPTH, and calcium absorption. The mean baseline 25(OH)D value for the entire sample was 70.0 nmol/L. Increases in 25(OH)D depended on dose with 12-week changes ranging from -10 nmol/L for placebo to 76 nmol/L for 4000 IU. Larger 25(OH)D gains were observed for whites vs blacks at the highest dose (P < .01). Gains for 1,25(OH)₂D were not significant (P = .07), and decreases in iPTH were not dose-dependent. There was no dose effect of vitamin D on fractional calcium absorption when adjusted for pill compliance, race, sex, or baseline 25(OH)D. Large increases in serum 25(OH)D with vitamin D₃ supplementation did not increase calcium absorption in healthy children living at 2 different latitudes. Supplementation with 400 IU/d was sufficient to maintain wintertime 25(OH)D concentrations in healthy black, but not white, children.


Framework fabrication with rib cartilage in partial and total nasal reconstruction.
The framework reconstruction of the nose is a significant and complex component of its partial or total reconstruction. On the one hand, the design of the individual framework parts is based on the anatomic nature of available rib or ear cartilage, which must on the other hand be adapted to the anatomic characteristics of the defect. The framework parts must be anchored not only to each other but also stably to the facial skeleton. The symmetry of the framework reconstruction is an essential component of the aesthetics of the reconstructed nose. If these points are already considered in planning, the reconstruction of the nasal framework can be standardized insofar as the same principles for the basic design of the individual parts as well as stable solutions for the anchoring points can be chosen. With reproducible techniques, functionally and aesthetically good to very good results can be achieved, including in the long term. The surgeon must possess special skills in the field of nasal reconstruction to correctly choose, apply, and combine the various techniques of nasal framework reconstruction.


Increased resonance energy transfer between fluorophores bound to DNA in proximity to metallic silver particles.
We examined the effects of metallic silver particles on resonance energy transfer (RET) between fluorophores covalently bound to DNA. A coumarin donor and a Cy3 acceptor were positioned at opposite ends of a 23-bp double helical DNA oligomer. In the absence of silver particles the extent of RET is near 9%, consistent with a Forster distance R(0) near 50 A and a donor to acceptor distance near 75 A. The transfer efficiency increased when the solution of AMCA-DNA-Cy3 was placed between two quartz plates coated with silver island films to near 64%, as determined by both steady-state and time-resolved measurements. The apparent R(0) in the presence of silver island films increases to about 110 A. These values of the transfer efficiency and R(0) represent weighted averages for donor-acceptor pairs near and distant from the metallic surfaces, so that the values at an optimal distance are likely to be larger. The increased energy transfer is observed only between two sandwiched silvered slides. When we replaced one silvered slide with a quartz plate the effect vanished. Also, the increased energy transfer was not observed for silvered slides separated more than a few micrometers. These results suggest the use of metal-enhanced RET in PCR, hybridization, and other DNA assays, and the possibility of controlling energy transfer by the distance between silver surfaces.


MR imaging technique for the diagnosis of pituitary iron overload in patients with transfusion-dependent beta-thalassemia major.
To identify the optimal MR imaging technique for diagnosing pituitary iron overload, we compared spin-echo and gradient-echo MR imaging with measurements of pituitary T2 relaxation times in 30 patients with secondary hemochromatosis due to transfusion-dependent beta-thalassemia major and in 10 healthy volunteers. We found that the optimal MR imaging technique to evaluate pituitary iron overload is the gradient-echo T2*-weighted technique, as it best demonstrated signal reduction in the anterior lobe of the pituitary gland.


Guided Imagery for Total Knee Replacement: Responses to an Audiobook Placebo Treatment.
The use of a placebo is the gold standard for establishing the efficacy of new treatments in randomized controlled trials (RCTs). Placebo treatments are inert; this allows specific effects of the experimental treatment to be separated from nonspecific physiological, psychological, and social effects. The aim of this study was to scrutinize the workings of an audiobook (AB) placebo treatment, which was used in a placebo-controlled, investigator-blinded, RCT of the effect of guided imagery (GI) to improve pain and function in persons undergoing total knee replacement. A secondary analysis of quantitative and narrative data collected during the RCT was performed. The AB treatment provided attention control. Participants in the AB group identified benefits-pleasant get-away, new knowledge, quiet time-out, and inspiration; however, there was also evidence of resentful demoralization, especially in the postoperative period. The use of ABs as a placebo treatment in GI RCTs should be executed with caution.


Serum ferritin as a predictor of host response to hepatitis B virus infection.
With hemodialysis patients, a high serum ferritin before there was serological evidence of hepatitis B virus infection increased the likelihood that the infection would be persistent. This finding suggested that hepatitis B virus is likely to infect and actively replicate in liver cells with the propensity for increased ferritin synthesis. The virus itself could stimulate the synthesis of ferritin in a cyclic positive feedback mechanism that increases intracellular ferritin concentration and, eventually, intracellular iron. Transformed liver cells have low iron content, do not replicate hepatitis B virus, and require iron for growth. Infected, nonmalignant liver cells could supply iron to the transformed cells and nourish their expansion.


Health Care utilization in elderly onset inflammatory bowel disease: a population-based study.
There is an increasing burden of inflammatory bowel disease (IBD) among the elderly. We sought to characterize health care utilization of elderly onset IBD. We identified incident IBD cases in Ontario, Canada between 1999 and 2008 and categorized subjects by age at diagnosis as young (18-40 yr), middle-age (41-64 yr), and elderly (≥65 yr). We compared IBD-specific health utilization indicators, including outpatient visits, emergency department visits, and hospitalizations. The elderly accounted for 8.1% (N = 725) and 11.6% (N = 1749) of incident Crohn's disease (CD) and ulcerative colitis (UC), respectively. They were less likely than young adults to have any IBD-specific gastroenterology visit in the first year after diagnosis (CD, 63% versus 71%, P < 0.001; UC, 63% versus 69%, P < 0.001). They less frequently received continuous gastroenterology care (CD, 36% versus 46%, P < 0.001; UC, 33% versus 43%, P < 0.001). Elderly patients with IBD were less likely than young adults to require an IBD-specific emergency department visit in the first year (CD, 8.8% versus 18.5%, P < 0.001; UC, 7.8% versus 11.6%, P < 0.001). Similarly, elderly patients with CD exhibited lower hospitalization rates (incidence rate ratio, 0.62; 95% confidence interval, 0.59-0.65). Hospitalization rates were modestly higher among those elderly patients with UC compared with young adults during the first year (incidence rate ratio, 1.14; 95% confidence interval: 1.02-1.28), but this association reversed thereafter (incidence rate ratio, 0.64; 95% confidence interval: 0.57-0.71). Elderly patients with IBD exhibited lower IBD-specific health care utilization than young adults, which may reflect a multitude of factors including more benign disease and differential health care access.


Benefits to poorly studied taxa of conservation of bird and mammal diversity on islands.
Protected area delineation and conservation action are urgently needed on marine islands, but the potential biodiversity benefits of these activities can be difficult to assess due to lack of species diversity information for lesser known taxa. We used linear mixed effects modeling and simple spatial analyses to investigate whether conservation activities based on the diversity of well-known insular taxa (birds and mammals) are likely to also capture the diversity of lesser known taxa (reptiles, amphibians, vascular land plants, ants, land snails, butterflies, and tenebrionid beetles). We assembled total, threatened, and endemic diversity data for both well-known and lesser known taxa and combined these with physical island biogeography characteristics for 1190 islands from 109 archipelagos. Among physical island biogeography factors, island area was the best indicator of diversity of both well-known and little-known taxa. Among taxonomic factors, total mammal species richness was the best indicator of total diversity of lesser known taxa, and the combination of threatened mammal and threatened bird diversity was the best indicator of lesser known endemic richness. The results of other intertaxon diversity comparisons were highly variable, however. Based on our results, we suggest that protecting islands above a certain minimum threshold area may be the most efficient use of conservation resources. For example, using our island database, if the threshold were set at 10 km(2) and the smallest 10% of islands greater than this threshold were protected, 119 islands would be protected. The islands would range in size from 10 to 29 km(2) and would include 268 lesser known species endemic to a single island, along with 11 bird and mammal species endemic to a single island. Our results suggest that for islands of equivalent size, prioritization based on total or threatened bird and mammal diversity may also capture opportunities to protect lesser known species endemic to islands.


WHAMM Directs the Arp2/3 Complex to the ER for Autophagosome Biogenesis through an Actin Comet Tail Mechanism.
Nucleation-promoting factors (NPFs) control the spatio-temporal activity of Arp2/3 complex in cells]. Thus, WASP and the WAVE complex direct the formation of branched actin networks at the leading edge during cell motility and endo/exocytosis, whereas the WASH complex is involved in endosomal transport. Less understood are WHAMM and JMY, two NPFs with similar domain architecture. JMY is found in the nucleus and the cytosol and is involved in transcriptional regulation, cell motility, and trans-Golgi transport. WHAMM was reported to bind microtubules and to be involved in ER to cis-Golgi transport. Here, we show that WHAMM directs the activity of Arp2/3 complex for autophagosome biogenesis through an actin-comet tail motility mechanism. Macroautophagy--the process by which cytosolic material is engulfed into autophagosomes for degradation and/or recycling--was recently shown to involve actin, but the mechanism is unknown. We found that WHAMM forms puncta that colocalize and comigrate with the autophagy markers LC3, DFCP1, and p62 through a WHAMM-dependent actin-comet tail mechanism. Under starvation, WHAMM and actin are observed at the interface between neighboring autophagosomes, whose number and size increase with WHAMM expression. Interfering with actin polymerization, inhibiting Arp2/3 complex, knocking down WHAMM, or blocking its interaction with Arp2/3 complex through mutagenesis all inhibit comet tail formation and reduce the size and number of autophagosomes. Finally, JMY shows similar localization to WHAMM and could be involved in similar processes. These results reveal a link between Arp2/3-complex-dependent actin assembly and autophagy.


[Wilson's Disease].
Wilson's disease is a rare inherited disorder of copper metabolism. If left untreated, the disorder has a fatal course within a few years after symptom onset. If discovered early, effective treatment is available, preventing further clinical deterioration and leading to a normal life expectancy. Accumulation of copper in liver and brain can lead to a variety of unspecific hepatic, neurologic and psychiatric symptoms such as jaundice, tremor, ataxia or depression. In patients under the age of 40 with unexplained hepatic, neurologic or psychiatric symptoms, Wilson's disease must be considered. Diagnosis can be made by measurement of urinary copper excretion and other tests as required. Lifelong decoppering treatment must be maintained. For the detection of deterioration of the disease or adverse effects of the medication as well as for the assessment of therapeutic compliance, regular clinical controls are necessary.


Long-term outcomes after coronary intervention with biodegradable polymer stents in patients with acute coronary syndromes.
Patients with acute coronary syndromes (ACS) may have worse outcomes after percutaneous coronary intervention compared to patients without ACS. To compare 5-year efficacy and safety outcomes in patients with and without ACS treated with biodegradable polymers, the ultrathin strut sirolimus-eluting Orsiro stent (O-SES) or the biolimus-eluting Nobori stent (N-BES). The Scandinavian Organisation for Randomized Trials with Clinical Outcome VII is a randomized trial comparing O-SES and N-BES in an all-comer setting. Of 2525 patients, 1329 (53%) patients had ACS and 1196 (47%) patients were without ACS. Endpoints were target lesion failure (TLF) (a composite of cardiac death, target lesion myocardial infarction, or target lesion revascularization) and definite stent thrombosis within 5 years. At 5-year follow-up, TLF did not differ significantly between patients with and without ACS (12.3% vs. 13.2%; rate ratio (RR) 1.00; 95% confidence interval (CI): 0.70-1.44), whereas the risk of definite stent thrombosis was increased in patients with ACS (2.3% vs. 1.3; RR: 2.01 [95% CI: 1.01-3.98]). In patients with ACS, the rate of TLF was similar between O-SES and N-BES (12.4% vs. 12.3%; RR: 1.02; 95% CI: 0.74-1.40). The reduced risk of definite stent thrombosis in O-SES treated ACS patients within the first year (0.2% vs. 1.6%; RR: 0.12; 95% CI: 0.02-0.93) was not maintained after 5 years (1.8% vs. 2.7%; RR: 0.77; 95% CI: 0.37-1.63). Patients with ACS had an increased risk of stent thrombosis regardless of the stent type used. Long-term outcomes were similar for ACS patients treated with O-SES or N-BES at 5 years.


Molecular basis of 5-hydroxytryptophan synthesis in Saccharomyces cerevisiae.
We report for the first time that 5-hydroxytryptophan can be synthesized in Saccharomyces cerevisiae by heterologously expressing prokaryotic phenylalanine 4-hydroxylase or eukaryotic tryptophan 3/5-hydroxylase, together with enhanced synthesis of MH4 or BH4 cofactors. The innate DFR1 gene in the folate synthesis pathway was found to play pivotal roles in 5-hydroxytryptophan synthesis.


Role-modeling and medical error disclosure: a national survey of trainees.
To measure trainees' exposure to negative and positive role-modeling for responding to medical errors and to examine the association between that exposure and trainees' attitudes and behaviors regarding error disclosure. Between May 2011 and June 2012, 435 residents at two large academic medical centers and 1,187 medical students from seven U.S. medical schools received anonymous, electronic questionnaires. The questionnaire asked respondents about (1) experiences with errors, (2) training for responding to errors, (3) behaviors related to error disclosure, (4) exposure to role-modeling for responding to errors, and (5) attitudes regarding disclosure. Using multivariate regression, the authors analyzed whether frequency of exposure to negative and positive role-modeling independently predicted two primary outcomes: (1) attitudes regarding disclosure and (2) nontransparent behavior in response to a harmful error. The response rate was 55% (884/1,622). Training on how to respond to errors had the largest independent, positive effect on attitudes (standardized effect estimate, 0.32, P < .001); negative role-modeling had the largest independent, negative effect (standardized effect estimate, -0.26, P < .001). Positive role-modeling had a positive effect on attitudes (standardized effect estimate, 0.26, P < .001). Exposure to negative role-modeling was independently associated with an increased likelihood of trainees' nontransparent behavior in response to an error (OR 1.37, 95% CI 1.15-1.64; P < .001). Exposure to role-modeling predicts trainees' attitudes and behavior regarding the disclosure of harmful errors. Negative role models may be a significant impediment to disclosure among trainees.


Targeting Stereotactic Body Radiotherapy on Metabolic PET- and Immuno-PET-Positive Vertebral Metastases.
(1) Background: Stereotactic body radiotherapy (SBRT) for vertebral metastases (VM) allows the delivery of high radiation doses to tumors while sparing the spinal cord. We report a new approach to clinical target volume (CTV) delineation based on anti-carcinoembryonic antigen (CEA) positron emission tomography (pretargeted immuno-PET; "iPET") in patients with metastatic breast cancer (BC) or medullary thyroid cancer (MTC). (2) Methods: All patients underwent iPET, spine magnetic resonance imaging (MRI), and positron emission tomography-computed tomography (PET-CT) using 18F-deoxyglucose (FDG) for BC or 18F-dihydroxy-phenylalanine (F-DOPA) for MTC. Vertebrae locations and vertebral segments of lesions were recorded and the impact on CTV delineation was evaluated. (3) Results: Forty-six VM eligible for SBRT following iPET were evaluated in eight patients (five BC, three MTC). Eighty-one vertebral segments were detected using MRI, 26 with FDG or F-DOPA PET/CT, and 70 using iPET. iPET was able to detect more lesions than MRI for vertebral bodies (44 vs. 34). iPET-based delineation modified MRI-based CTV in 70% (32/46) of cases. (4) Conclusion: iPET allows a precise mapping of affected VM segments, and adds complementary information to MRI in the definition of candidate volumes for VM SBRT. iPET may facilitate determining target volumes for treatment with stereotactic body radiotherapy in metastatic vertebral disease.


Searching for biomarkers of CDKL5 disorder: early-onset visual impairment in CDKL5 mutant mice.
CDKL5 disorder is a neurodevelopmental disorder still without a cure. Murine models of CDKL5 disorder have been recently generated raising the possibility of preclinical testing of treatments. However, unbiased, quantitative biomarkers of high translational value to monitor brain function are still missing. Moreover, the analysis of treatment is hindered by the challenge of repeatedly and non-invasively testing neuronal function. We analyzed the development of visual responses in a mouse model of CDKL5 disorder to introduce visually evoked responses as a quantitative method to assess cortical circuit function. Cortical visual responses were assessed in CDKL5 null male mice, heterozygous females, and their respective control wild-type littermates by repeated transcranial optical imaging from P27 until P32. No difference between wild-type and mutant mice was present at P25-P26 whereas defective responses appeared from P27-P28 both in heterozygous and homozygous CDKL5 mutant mice. These results were confirmed by visually evoked potentials (VEPs) recorded from the visual cortex of a different cohort. The previously imaged mice were also analyzed at P60-80 using VEPs, revealing a persistent reduction of response amplitude, reduced visual acuity and defective contrast function. The level of adult impairment was significantly correlated with the reduction in visual responses observed during development. Support vector machine showed that multi-dimensional visual assessment can be used to automatically classify mutant and wt mice with high reliability. Thus, monitoring visual responses represents a promising biomarker for preclinical and clinical studies on CDKL5 disorder.


Hormone-induced protein phosphorylation. II. Localization to the ribosomal fraction from rat exocrine pancreas and parotid of a 29,000-dalton protein phosphorylated in situ in response to secretagogues.
In the preceding paper, we demonstrated that the endogenous phosphorylation of a protein with a molecular weight of 29,000 was enhanced by various secretagogues in rat pancreatic and parotid lobules, the phosphorylation of this protein correlating both temporally and in a dose-dependent fashion with secretory protein discharge. In the present study, we established a specific methodology to characterize this phosphoprotein. Once established, this 29,000-dalton phosphoprotein was then followed selectively and quantitatively throughout subcellular fractionation procedures. Analysis of two-dimensional polyacrylamide gels demonstrated that proteins with similar mobilities (Mr 29,000; pl greater than 8.4) were affected by cholecystokinin octapeptide and isoproterenol in rat pancreatic and parotid lobules, respectively, suggesting that the same 29,000-dalton phosphoprotein was covalently modified in both tissues. Cellular fractionation studies using differential velocity and sucrose density gradient centrifugation revealed that the 29,000-dalton phosphoprotein copurified with the rough microsomal fraction of pancreas and was highly enriched in ribosomal fractions of both pancreas and parotid. Electrophoresis in two dimensions confirmed that the 29,000-dalton polypeptide that was resolved directly from stimulated cells and from ribosomal fractions exhibited a common mobility, and apparent identity of the species was strongly suggested when the 29,000-dalton polypeptides from both sources were compared by peptide mapping following limited digestion with Staphylococcus aureus V8 protease. This phosphoprotein was tentatively identified as ribosomal protein S6 after analysis by pH 8.6/4.2 two-dimensional PAGE.


Bioactivity of ethanol extract of Karanja (Pongamia glabra vent) seed coat against mosquitoes.
Laboratory evaluation revealed that the treatment of larvae of Aedes aegypti and Culex quinquefasciatus with ethanol extract of karanja seed coat (ALKSC) significantly increased the larval mortality and developmental period proportianately with increase in the extract concentrations. Aedes proved more sensitive to the effect of extract in terms of mortality than Culex. At a high concentration (8 ppm), 100% Aedes larvae died in the first instar within two days whereas, 16 pmm was required for achieving the same level of mortality in Culex. Culex was also found more sensitive than Aedes regarding IGR effect. In emerging Culex pupae several abnormalities were observed while Aedes pupae did not exhibit any abnormalities. However larvae which pupated successfully resulted in adults with several structural abnormalities in both the species.


Pectinous cell wall thickenings formation - A common defense strategy of plants to cope with Pb.
Lead, one of the most abundant and hazardous trace metals affecting living organisms, has been commonly detected in plant cell walls including some tolerant plants, mining ecotypes and hyperaccumulators. We have previously shown that in tip growing Funaria sp. protonemata cell wall is remodeled in response to lead by formation of thickenings rich in low-methylesterified pectins (pectin epitope JIM5 - JIM5-P) able to bind metal ions, which accumulate large amounts of Pb. Hence, it leads to the increase of cell wall capacity for Pb compartmentalization. Here we show that diverse plant species belonging to different phyla (Arabidopsis, hybrid aspen, star duckweed), form similar cell wall thickenings in response to Pb. These thickenings are formed in tip growing cells such as the root hairs, and in diffuse growing cells such as meristematic and root cap columella cells of root apices in hybrid aspen and Arabidopsis and in mesophyll cells in star duckweed fronds. Notably, all analyzed cell wall thickenings were abundant in JIM5-P and accumulated high amounts of Pb. In addition, the co-localization of JIM5-P and Pb commonly occurred in these cells. Hence, cell wall thickenings formed the extra compartment for Pb accumulation. In this way plant cells increased cell wall capacity for compartmentalization of this toxic metal, protecting protoplast from its toxicity. As cell wall thickenings occurred in diverse plant species and cell types differing in the type of growth we may conclude that pectinous cell wall thickenings formation is a widespread defense strategy of plants to cope with Pb. Moreover, detection of natural defense strategy, increasing plant cell walls capacity for metal accumulation, reveals a promising direction for enhancing plant efficiency in phytoremediation.


Congenital abnormalities of the breast.
Congenital abnormalities of the breast and chest wall are seen frequently in breast, pediatric and plastic surgery clinics. Management involves a multidisciplinary team approach. The treatment for many of these conditions includes surgical correction. If the child is still growing, treatment timing is crucial and many of the surgical corrective procedures require more than one operation over a number of years. Although these deformities have less impact on functional capacity, the psychological consequences can be serious in adolescent patients. They can experience embarrassment, social isolation and complexities during sexual development, and this results in problems with relationships. The aim of this article is to spread awareness among the medical community of this range of conditions, so appropriate referrals are made and their impact is reduced by timely appropriate surgical intervention if appropriate.


The differences in feeding-inhibitory responses to peripheral and central leptin between non-lactating and lactating rats.
This study was conducted to examine the contributions of central and peripheral leptin to hyperphagia in lactation. Lactating rats were mated at 7-8 weeks of age and housed singly with their litters. In experiment 1, food intakes were significantly (P<0.01) greater (350% on average) in lactation than in non-lactation throughout a day. Cerebrospinal fluid (CSF) leptin levels remained constant despite plasma leptin levels being significantly (P<0.05) greater in non-lactation than in lactation. In experiment 2, CSF leptin levels were not altered by i.v. injections of leptin (0.2 and 0.4 mg/kg body weight) despite that plasma leptin levels were dose dependently (P<0.01) increased. Moreover, i.v. administration of leptin significantly (P<0.05) decreased food intake in non-lactating rats but not in lactating rats. In experiment 3, nocturnal food intakes were temporarily (P<0.05) reduced in non-lactating and lactating rats. I.c.v. administration of a leptin antagonist (15 μg) blocked the reductions of food intakes. I.c.v. administration of leptin (10 μg) significantly (P<0.05) decreased cumulative food intakes during 24 h in both the physiological states. In conclusion, this study has presented new evidence that the hyperphagia of lactating rats could be partly due to depressed sensitivity of neurons contacting blood leptin. In contrast, the responsiveness of leptin receptors contacting CSF leptin may not differ between non-lactating and lactating rats. Furthermore, the levels of CSF leptin remained constant independent of those of blood leptin. Therefore, the expression of hypothalamic leptin receptors contacting CSF could be involved in the difference in food intake between non-lactating and lactating rats.


Doxycycline attenuates burn-induced microvascular hyperpermeability.
Burns induce systemic microvascular hyperpermeability resulting in shock, and if untreated, cardiovascular collapse. Damage to the endothelial cell adherens junctional complex plays an integral role in the pathophysiology of microvascular hyperpermeability. We hypothesized that doxycycline, a known inhibitor of matrix metalloproteinases (MMPs), could attenuate burn-induced adherens junction damage and microvascular hyperpermeability. Male Sprague-Dawley rats were divided into sham, burn, and burn + doxycycline (n = 5). The experimental groups underwent a 30% total body surface area full-thickness burn. Fluorescein isothiocyanate-albumin was administered intravenously. Mesenteric postcapillary venules were examined with intravital microscopy to determine flux of albumin from the intravascular space to the interstitium. Fluorescence intensity was compared between the intravascular space to the interstitium at 30, 60, 80, 100, 120, 140, 160, and 180 minutes after burn. Parallel experiments were performed in which rat lung microvascular endothelial cells were treated with sera from sham or burn animals as well as separate groups pretreated with either doxycycline or a specific inhibitor of MMP-9. Monolayer permeability was determined by fluorescein isothiocyanate albumin-flux across Transwell plates and immunofluorescense staining for the adherens junction protein β-catenin was performed. Western blot and gelatin zymography were performed to assess MMP-9 level and activity. MMP-9 levels were increased after burn. Monolayer permeability was significantly increased with burn serum treatment; this was attenuated with doxycycline as well as the specific MMP-9 inhibitor (p < 0.05). Damage of the endothelial cell adherens junction complex was induced by serum from burned rats, and doxycycline restored the integrity of the adherens junction similar to the MMP-9 inhibitor. Intravital microscopy revealed microvascular hyperpermeability after burn; this was attenuated with doxycycline (p < 0.05). Burns induce microvascular hyperpermeability via endothelial adherens junction disruption associated with MMP-9, and this is attenuated with doxycycline.


Impact of water regimes on an experimental community of four desert arbuscular mycorrhizal fungal (AMF) species, as affected by the introduction of a non-native AMF species.
Field studies have revealed the impact of changing water regimes on the structure of arbuscular mycorrhizal fungal (AMF) communities, but it is not known what happens to the abundance of individual AMF species within the community when the water conditions in the rhizosphere change. The behavior of four AMF species isolated from the Arabian desert (Diversispora aurantia, Diversispora omaniana, Septoglomus africanum, and an undescribed Paraglomus species) was investigated when assembled in microcosms containing Sorghum bicolor as host plant, and treated with various water regimes. Furthermore, the impact of invasion of these assemblages by Rhizophagus irregularis, an AMF species widely used in commercial inocula, was studied. The abundance of each AMF species in sorghum roots was measured by determining the transcript numbers of their large ribosomal subunit (rLSU) by real-time PCR, using cDNA and species-specific primers. Plant biomass and length of AMF extraradical hyphae were also measured. The abundance of each AMF species within the sorghum roots was influenced by both the water regime and the introduction of R. irregularis. Under dry conditions, the introduction of R. irregularis reduced the total abundance of all native AMF species in roots and also led to a reduction in the amount of extraradical mycelium, as well as to a partial decrease in plant biomass. The results indicate that both water regime and the introduction of an invasive AMF species can strongly alter the structure of an AMF native assemblage with a consequent impact on the entire symbiotic mycorrhizal relationship.


Empirical evidence of bias in the design of experimental stroke studies: a metaepidemiologic approach.
At least part of the failure in the transition from experimental to clinical studies in stroke has been attributed to the imprecision introduced by problems in the design of experimental stroke studies. Using a metaepidemiologic approach, we addressed the effect of randomization, blinding, and use of comorbid animals on the estimate of how effectively therapeutic interventions reduce infarct size. Electronic and manual searches were performed to identify meta-analyses that described interventions in experimental stroke. For each meta-analysis thus identified, a reanalysis was conducted to estimate the impact of various quality items on the estimate of efficacy, and these estimates were combined in a meta-meta-analysis to obtain a summary measure of the impact of the various design characteristics. Thirteen meta-analyses that described outcomes in 15,635 animals were included. Studies that included unblinded induction of ischemia reported effect sizes 13.1% (95% CI, 26.4% to 0.2%) greater than studies that included blinding, and studies that included healthy animals instead of animals with comorbidities overstated the effect size by 11.5% (95% CI, 21.2% to 1.8%). No significant effect was found for randomization, blinded outcome assessment, or high aggregate CAMARADES quality score. We provide empirical evidence of bias in the design of studies, with studies that included unblinded induction of ischemia or healthy animals overestimating the effectiveness of the intervention. This bias could account for the failure in the transition from bench to bedside of stroke therapies.


A subspace reverse-correlation technique for the study of visual neurons.
A new discrete-time reverse-correlation scheme for the study of visual neurons is proposed. The visual stimulus is generated by drawing with uniform probability, at each refresh time, an image from a finite set S of orthonormal images. We show that if the neuron can be modeled as a spatiotemporal linear filter followed by a static nonlinearity, the cross-correlation between the input image sequence and the cell's spike train output gives the projection of the receptive field onto the subspace spanned by S. The technique has been applied to the analysis of simple cells in the primary visual cortex of cats and macaque monkeys. Experimental results are presented where S spans a subspace of spatially low-pass signals. Advantages of the proposed scheme over standard white-noise techniques include improved signal to noise ratios, increased spatial resolution, and the possibility to restrict the study to particular subspaces of interest.


TNF-alpha induction by nickel compounds is specific through ERKs/AP-1-dependent pathway in human bronchial epithelial cells.
The chronic lung inflammatory activity and carcinogenicity of nickel compounds have been well documented by previous studies from epidemiology both in vitro and in vivo. However, the molecular mechanism involved in nickel-induced chronic lung inflammation is much less understood. The current study demonstrates that exposure of human bronchial epithelial cells (Beas-2B) to nickel compounds results in the induction of the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) and transactivation of nuclear factor of activated T cells (NFAT), nuclear factor-kappaB (NF-kappaB), and activator protein-1 (AP-1). Further studies show that neither overexpression of IKKbeta-KM, a kinase inactive mutant of IKKbeta, nor the ectopic expression of a dominant negative mutant of NFAT could inhibit the TNF-alpha induction by nickel exposure. Overexpression of TAM67, a dominant-negative mutant of c-Jun, dramatically reduced the TNF-alpha induction, suggesting that AP-1 is a mediator of TNF-alpha induction in nickel responses. Our results show that ERKs are AP-1 upstream kinases responsible for TNF-alpha induction by nickel exposure; although JNKs, ERKs, and p38K were all activated in the Beas-2B cells exposed to nickel compounds. Our results demonstrate that inflammatory TNF-alpha could be induced by nickel exposure in Beas-2B cells specifically through an ERKs/AP-1-dependent pathway.


Identification of valproic acid glucuronide hydrolase as a key enzyme for the interaction of valproic acid with carbapenem antibiotics.
Plasma levels of valproic acid (VPA) are decreased by concomitant use with carbapenem antibiotics, such as panipenem (PAPM). One of the plausible mechanisms of this interaction is the inhibition of VPA glucuronide (VPA-G) hydrolysis by carbapenems in the liver. To elucidate this interaction mechanism, we purified VPA-G hydrolase from human liver cytosol, in which the hydrolytic activity was mainly located. After chromatographic purification, the VPA-G hydrolase was identified as acylpeptide hydrolase (APEH). APEH-depleted cytosol, prepared by an immunodepletion method, completely lacked the hydrolytic activity. These results demonstrate that APEH is a single enzyme involved in PAPM-sensitive VPA-G hydrolysis in cytosol. In addition, the hydrolytic activity of recombinant human APEH was inhibited by PAPM and the inhibition profile by typical esterase inhibitors (diisopropyl fluorophosphate, 5,5'-dithiobis(2-nitrobenzoic acid), p-chloromercuribenzoic acid, and d-saccharic acid 1,4-lactone) was similar to that of human liver cytosol. Cytosolic VPA-G hydrolase activity was slightly inhibited by cholinesterase and carboxylesterase inhibitors. beta-Glucuronidase activity remained in APEH-depleted cytosol, whereas VPA-G hydrolase activity was completely abolished. Thus, either cholinesterase, carboxylesterase, or beta-glucuronidase in cytosol would not be involved in VPA-G hydrolysis. Taken together, APEH plays a major role in the PAPM-sensitive VPA-G hydrolysis in the liver. These findings suggest that APEH could be a key enzyme for the drug interaction of VPA with carbapenems via VPA-G hydrolysis.