Increase in African dust flux at the onset of commercial agriculture in the Sahel region.
The Sahara Desert is the largest source of mineral dust in the world. Emissions of African dust increased sharply in the early 1970s (ref. 2), a change that has been attributed mainly to drought in the Sahara/Sahel region caused by changes in the global distribution of sea surface temperature. The human contribution to land degradation and dust mobilization in this region remains poorly understood, owing to the paucity of data that would allow the identification of long-term trends in desertification. Direct measurements of airborne African dust concentrations only became available in the mid-1960s from a station on Barbados and subsequently from satellite imagery since the late 1970s: they do not cover the onset of commercial agriculture in the Sahel region approximately 170 years ago. Here we construct a 3,200-year record of dust deposition off northwest Africa by investigating the chemistry and grain-size distribution of terrigenous sediments deposited at a marine site located directly under the West African dust plume. With the help of our dust record and a proxy record for West African precipitation we find that, on the century scale, dust deposition is related to precipitation in tropical West Africa until the seventeenth century. At the beginning of the nineteenth century, a sharp increase in dust deposition parallels the advent of commercial agriculture in the Sahel region. Our findings suggest that human-induced dust emissions from the Sahel region have contributed to the atmospheric dust load for about 200 years.


Outbreak of Klebsiella pneumoniae carbapenemase-2-producing K. pneumoniae sequence type 11 in Taiwan in 2011.
From June to September 2011, a total of 305 ertapenem-nonsusceptible Enterobacteriaceae isolates (MICs of ertapenem ≥ 1 μg/ml) were collected from 11 hospitals in different parts of Taiwan. The MICs of 12 antimicrobial agents against these isolates were determined using the broth microdilution method, and genes for carbapenemases were detected using PCR. Genotypes of isolates possessing carbapenemase genes were identified by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. The ertapenem-nonsusceptible Enterobacteriaceae isolates included Klebsiella pneumoniae (n = 219), Escherichia coli (n = 64), Enterobacter cloacae (n = 15), and other species (n = 7). Seven (2.3%) of the ertapenem-nonsusceptible Enterobacteriaceae isolates exhibited colistin MICs of >4 μg/ml, and 24 (7.9%) were not susceptible to tigecycline (MICs > 2 μg/ml). A total of 29 (9.5%) isolates carried genes encoding carbapenemases, namely, K. pneumoniae carbapenemase-2 (KPC-2) in 16 (7.3%) isolates of K. pneumoniae (KPC-2-KP) and IMP-8 in 5 (2.3%) isolates of K. pneumoniae, 5 (33.3%) isolates of E. cloacae, 1 isolate of E. coli, 1 isolate of Klebsiella oxytoca, and one isolate of Citrobacter freundii. The 16 KPC-2-KP isolates were isolated from patients at four different hospitals in northern Taiwan. All 16 of the KPC-2-KP isolates were susceptible to amikacin and colistin and had a similar pulsotype (pulsotype 1) and the same sequence type (sequence type 11). Infections due to KPC-2-KP mainly occurred in severely ill patients in the intensive care unit (n = 14, 88%). Four patients with infections due to KPC-2-KP died within 14 days of hospitalization. The findings are the first to demonstrate intrahospital and interhospital dissemination of KPC-2-KP in northern Taiwan.


The Ramazzini Institute 13-week pilot study on glyphosate and Roundup administered at human-equivalent dose to Sprague Dawley rats: effects on the microbiome.
Glyphosate-based herbicides (GBHs) are broad-spectrum herbicides that act on the shikimate pathway in bacteria, fungi, and plants. The possible effects of GBHs on human health are the subject of an intense public debate for both its potential carcinogenic and non-carcinogenic effects, including its effects on microbiome. The present pilot study examines whether exposure to GBHs at doses of glyphosate considered to be "safe" (the US Acceptable Daily Intake - ADI - of 1.75 mg/kg bw/day), starting from in utero, may modify the composition of gut microbiome in Sprague Dawley (SD) rats. Glyphosate alone and Roundup, a commercial brand of GBHs, were administered in drinking water at doses comparable to the US glyphosate ADI (1.75 mg/kg bw/day) to F0 dams starting from the gestational day (GD) 6 up to postnatal day (PND) 125. Animal feces were collected at multiple time points from both F0 dams and F1 pups. The gut microbiota of 433 fecal samples were profiled at V3-V4 region of 16S ribosomal RNA gene and further taxonomically assigned and assessed for diversity analysis. We tested the effect of exposure on overall microbiome diversity using PERMANOVA and on individual taxa by LEfSe analysis. Microbiome profiling revealed that low-dose exposure to Roundup and glyphosate resulted in significant and distinctive changes in overall bacterial composition in F1 pups only. Specifically, at PND31, corresponding to pre-pubertal age in humans, relative abundance for Bacteriodetes (Prevotella) was increased while the Firmicutes (Lactobacillus) was reduced in both Roundup and glyphosate exposed F1 pups compared to controls. This study provides initial evidence that exposures to commonly used GBHs, at doses considered safe, are capable of modifying the gut microbiota in early development, particularly before the onset of puberty. These findings warrant future studies on potential health effects of GBHs in early development such as childhood.


When Is a Test Score Fair for the Individual Who Is Being Tested? Effects of Different Scoring Procedures across Multiple Attempts When Testing a Motor Skill Task.
Tests or test batteries used for assessing motor skills, either in research studies or in clinical settings, apply a variety of procedures for scoring performances, including everything from one to ten attempts, of which the best is scored or an average is computed. The rationale behind scoring procedures is rarely stated, and it seems that the number of attempts allowed is decided without much qualification from research. It is uncertain whether procedures fairly capture an individual's skill level. Thus, the validity of the tests may be compromised. The present study tested 24 young female soccer players on the juggling of a soccer ball. They were given 10 attempts, and trials were scored according to nine different procedures including the 'best of' or 'mean of' either one, two, three, five, or ten attempts. Individual raw scores differed widely across trials, but no general effect of trials was found. The mean (SD) percentage difference between the lowest and highest scores was 27.7(9.9)%, with 17 players (71%) demonstrating a significant change from lowest to highest score. Correlations between raw scores were low across trials, while they were generally higher across scoring procedures. The first trial was significantly different from the remaining both as a raw score and as scoring procedure. The mean percentage difference between best-of-two and best-of-ten scores was 95%, with 50 % of the players demonstrating a significant difference between the two scoring procedures. No significant differences were found across mean-of-rule scorings. Best-of-rule and mean-of-rule scorings were significantly different except for the best-of-two vs. mean-of-two. The mean difference between highest and lowest rank across players was 6.7 (3.6), with individual rankings within the group varying 33% on average across procedures. One player moved from 3rd to 23rd place because of procedural differences. Therefore, it is concluded that scoring procedures affect results and may have an impact on test outcomes. This may present consequences for decision-making from test results, such as diagnosing and selection of intervention groups. We hope that our results would inspire further research into the scoring procedures of the vast amount of tests and tasks in common use.


Reconstruction of large, irregularly sampled multidimensional images. A tensor-based approach.
Many practical applications require the reconstruction of images from irregularly sampled data. The spline formalism offers an attractive framework for solving this problem; the currently available methods, however, are hard to deploy for large-scale interpolation problems in dimensions greater than two (3-D, 3-D+time) because of an exponential increase of their computational cost (curse of dimensionality). Here, we revisit the standard regularized least-squares formulation of the interpolation problem, and propose to perform the reconstruction in a uniform tensor-product B-spline basis as an alternative to the classical solution involving radial basis functions. Our analysis reveals that the underlying multilinear system of equations admits a tensor decomposition with an extreme sparsity of its one dimensional components. We exploit this property for implementing a parallel, memory-efficient system solver. We show that the computational complexity of the proposed algorithm is essentially linear in the number of measurements and that its dependency on the number of dimensions is significantly less than that of the original sparse matrix-based implementation. The net benefit is a substantial reduction in memory requirement and operation count when compared to standard matrix-based algorithms, so that even 4-D problems with millions of samples become computationally feasible on desktop PCs in reasonable time. After validating the proposed algorithm in 3-D and 4-D, we apply it to a concrete imaging problem: the reconstruction of medical ultrasound images (3-D+time) from a large set of irregularly sampled measurements, acquired by a fast rotating ultrasound transducer.


The use of multiple-criteria decision-making theory to measure students' perceptions of high-fidelity simulation.
High-Fidelity Simulation (HFS) has great potential to improve decision-making in clinical practice. Previous studies have found HFS promotes self-confidence, but its effectiveness in clinical practice has not been established. The aim of this research is to establish if HFS facilitates learning that informs decision-making skills in clinical practice using MultipleCriteria DecisionMaking Theory (MCDMT). The sample was 2nd year undergraduate pre-registration adult nursing students.MCDMT was used to measure the students' experience of HFS and how it developed their clinical decision-making skills. MCDMT requires characteristic measurements which for the learning experience were based on five factors that underpin successful learning, and for clinical decision-making, an analytical framework was used. The study used a repeated-measures design to take two measurements: the first one after the first simulation experience and the second one after clinical placement. Baseline measurements were obtained from academics. Data were analysed using the MCDMT tool. After their initial exposure to simulation learning, students reported that HFS provides a high-quality learning experience (87%) and supports all aspects of clinical decision-making (85%). Following clinical practice, the level of support for clinical decision-making remained at 85%, suggesting that students believe HFS promotes transferability of knowledge to the practice setting. Overall, students report a high level of support for learning and developing clinical decision-making skills from HFS. However, there are no comparative data available from classroom teaching of similar content so it cannot be established if these results are due to HFS alone.


Enhancing effects of anti-CD40 treatment on the immune response of SCID-bovine mice to Trypanosoma congolense infection.
African trypansosomes are tsetse-transmitted parasites of chief importance in causing disease in livestock in regions of sub-Saharan Africa. Previous studies have demonstrated that certain breeds of cattle are relatively resistant to infection with trypanosomes, and others are more susceptible. Because of its extracellular location, the humoral branch of the immune system dominates the response against Trypanosoma congolense. In the following study, we describe the humoral immune response generated against T. congolense in SCID mice reconstituted with a bovine immune system (SCID-bo). SCID-bo mice infected with T. congolense were treated with an agonistic anti-CD40 antibody and monitored for the development of parasitemia and survival. Anti-CD40 antibody administration resulted in enhanced survival compared with mice receiving the isotype control. In addition, we demonstrate that the majority of bovine IgM+ B cells in SCID-bo mice expresses CD5, consistent with a neonatal phenotype. It is interesting that the percentage of bovine CD5+ B cells in the peripheral blood of infected SCID-bo mice was increased following anti-CD40 treatment. Immunohistochemical staining also indicated increased numbers of Ig+ cells in the spleens of anti-CD40-treated mice. Consistent with previous studies demonstrating high IL-10 production during high parasitemia levels in mice and cattle, abundant IL-10 mRNA message was detected in the spleens and peripheral blood of T. congolense-infected SCID-bo mice during periods of high parasitemia. In addition, although detected in plasma when parasites were absent or low in number, bovine antibody was undetectable during high parasitemia. However, Berenil treatment allowed for the detection of VSG-specific IgG 14 days postinfection in T. congolense-infected SCID-bo mice. Overall, the data indicate that survival of trypanosome-infected SCID-bo mice is prolonged when an agonistic antibody against bovine CD40 (ILA156) is administered. Thus, stimulation of B cells and/or other cell types through CD40 afforded SCID-bo mice a slight degree of protection during T. congolense infection.


Target dose conversion modeling from pencil beam (PB) to Monte Carlo (MC) for lung SBRT.
A challenge preventing routine clinical implementation of Monte Carlo (MC)-based lung SBRT is the difficulty of reinterpreting historical outcome data calculated with inaccurate dose algorithms, because the target dose was found to decrease to varying degrees when recalculated with MC. The large variability was previously found to be affected by factors such as tumour size, location, and lung density, usually through sub-group comparisons. We hereby conducted a pilot study to systematically and quantitatively analyze these patient factors and explore accurate target dose conversion models, so that large-scale historical outcome data can be correlated with more accurate MC dose without recalculation. Twenty-one patients that underwent SBRT for early-stage lung cancer were replanned with 6MV 360° dynamic conformal arcs using pencil-beam (PB) and recalculated with MC. The percent D95 difference (PB-MC) was calculated for the PTV and GTV. Using single linear regression, this difference was correlated with the following quantitative patient indices: maximum tumour diameter (MaxD); PTV and GTV volumes; minimum distance from tumour to soft tissue (dmin); and mean density and standard deviation of the PTV, GTV, PTV margin, lung, and 2 mm, 15 mm, 50 mm shells outside the PTV. Multiple linear regression and artificial neural network (ANN) were employed to model multiple factors and improve dose conversion accuracy. Single linear regression with PTV D95 deficiency identified the strongest correlation on mean-density (location) indices, weaker on lung density, and the weakest on size indices, with the following R(2) values in decreasing orders: shell2mm (0.71), PTV (0.68), PTV margin (0.65), shell15mm (0.62), shell50mm (0.49), lung (0.40), dmin (0.22), GTV (0.19), MaxD (0.17), PTV volume (0.15), and GTV volume (0.08). A multiple linear regression model yielded the significance factor of 3.0E-7 using two independent features: mean density of shell2mm (P = 1.6E-7) and PTV volume (P = 0.006). A 4-feature ANN model slightly improved the modeling accuracy. Quantifiable density features were proposed, replacing simple central/peripheral location designation, which showed strong correlations with PB-to-MC target dose conversion magnitude, followed by lung density and target size. Density in the immediate outer and inner areas of the PTV showed the strongest correlations. A multiple linear regression model with one such feature and PTV volume established a high significance factor, improving dose conversion accuracy.


Microthrombus formation may trigger lung injury after acute blood loss.
We showed previously that acute blood loss, without resuscitation, caused marked maldistribution of interalveolar perfusion. Because hemorrhage is a known risk factor for the development of lung injury, the goal of our present studies was to determine if there was a correlation between perfusion maldistribution and the subsequent development of lung injury after blood loss. Specifically, we wanted to know if the perfusion maldistribution might be due to microthrombus formation and/or leukocyte sequestration within the pulmonary microcirculation. We bled rats (30% blood loss) and harvested their lungs 45 min or 24 h later. Lungs were prepared for perfusion distribution analysis, Western blot analysis to measure whole-lung fibrinogen concentrations, and for immunohistochemistry to measure fibrin deposition and leukocyte deposition (CD16 fluorescence). Perfusion was significantly maldistributed at 45 min and 24 h (P < 0.05). At 45 min, whole-lung fibrinogen concentrations were less than half that in controls (P < 0.05), whereas numbers of fibrin microthrombi were 2.5-fold greater than control by 45 min (not statistically significant) and were 4.5-fold greater by 24 h (P = 0.01). Leukocyte deposition was two-fold greater than control by 45 min (not statistically significant) and was 4-fold greater by 24 h (P = 0.02). Fibrin-to-leukocyte nearest-neighbor distances remained unchanged (18.1 [SD, 1.1] μm) even as the numbers of both increased with time after blood loss. Our results suggest that soluble fibrinogen polymerized to insoluble fibrin within minutes after acute blood loss, which caused perfusion maldistribution and attracted leukocytes. The development of lung injury after blood loss may be a consequence of leukocyte chemoattraction to fibrin microthrombi that seem to form within minutes after blood loss.


Prenatal alcohol exposure and childhood behavior at age 6 to 7 years: I. dose-response effect.
Moderate to heavy levels of prenatal alcohol exposure have been associated with alterations in child behavior, but limited data are available on adverse effects after low levels of exposure. The objective of this study was to evaluate the dose-response effect of prenatal alcohol exposure for adverse child behavior outcomes at 6 to 7 years of age. Beginning in 1986, women attending the urban university-based maternity clinic were routinely screened at their first prenatal visit for alcohol and drug use by trained research assistants from the Fetal Alcohol Research Center. All women reporting alcohol consumption at conception of at least 0.5 oz absolute alcohol/day and a 5% random sample of lower level drinkers and abstainers were invited to participate to be able to identify the associations between alcohol intake and child development. Maternal alcohol, cigarette, and illicit drug use were prospectively assessed during pregnancy and postnatally. The independent variable in this study, prenatal alcohol exposure, was computed as the average absolute alcohol intake (oz) per day across pregnancy. At each prenatal visit, mothers were interviewed about alcohol use during the previous 2 weeks. Quantities and types of alcohol consumed were converted to fluid ounces of absolute alcohol and averaged across visits to generate a summary measure of alcohol exposure throughout pregnancy. Alcohol was initially used as a dichotomous variable comparing children with no prenatal alcohol exposure to children with any exposure. To evaluate the effects of different levels of exposure, the average absolute alcohol intake was relatively arbitrarily categorized into no, low (>0 but <0.3 fl oz of absolute alcohol/day), and moderate/heavy (>/=0.3 fl oz of absolute alcohol/day) for the purpose of this study. Six years later, 665 families were contacted. Ninety-four percent agreed to testing. Exclusions included children who missed multiple test appointments, had major congenital malformations (other than fetal alcohol syndrome), possessed an IQ >2 standard deviations from the sample mean, or had incomplete data. The Achenbach Child Behavior Checklist (CBCL) was used to assess child behavior. The CBCL is a parent questionnaire applicable to children ages 4 to 16 years. It is widely used in the clinical assessment of children's behavior problems and has been extensively used in research. Eight syndrome scales are further grouped into Externalizing or undercontrolled (Aggressive and Delinquent) behavior and Internalizing or overcontrolled (Anxious/Depressed, Somatic Complaints, and Withdrawn) behaviors. Three syndromes (Social, Thought, and Attention Problems) fit neither group. Higher scores are associated with more problem behaviors. Research assistants who were trained and blinded to exposure status independently interviewed the child and caretaker. Data were collected on a broad range of control variables known to influence childhood behavior and/or to be associated with prenatal alcohol exposure. These included perinatal factors of maternal age, education, cigarette, cocaine, and other substances of abuse and the gestational age of the baby. Postnatal factors studied included maternal psychopathology, continuing alcohol and drug use, family structure, socioeconomic status, children's whole blood lead level, and exposure to violence. Data were collected only from black women as there was inadequate representation of other racial groups. Statistical analyses were performed using the SPSS statistical package. Frequency distribution, cross-tabulation, odds ratio, and chi(2) tests were used for analyzing categorical data. Continuous data were analyzed using t tests, analyses of variance (ANOVAs) with posthoc tests, and regression analysis. Testing was available for 501 parent-children dyads. Almost one fourth of the women denied alcohol use during pregnancy. Low levels of alcohol use were reported in 63.8% and moderate/heavy use in 13% of pregnancies. Increasing prenatal alcohol exposure was associated with lower birth weight and gestational age, higher lead levels, higher maternal age, and lower education level, prenatal exposure to cocaine and smoking, custody changes, lower socioeconomic status, and paternal drinking and drug use at the time of pregnancy. Children with any prenatal alcohol exposure were more likely to have higher CBCL scores on Externalizing (Aggressive and Delinquent) and Internalizing (Anxious/Depressed and Withdrawn) syndrome scales and the Total Problem Score. The odds ratio of scoring in the clinical range for Delinquent behavior was 3.2 (1.3-7.6) in children with any prenatal exposure to alcohol compared with nonexposed controls. The threshold dose was evaluated with the 3 prenatal alcohol exposure groups. One-way ANOVA revealed a significant between group difference for Externalizing (Aggressive and Delinquent) and the Total Problem Score. (ABSTRACT TRUNCATED)


A Boron Difluoride Hydrazone (BODIHY) Polymer Exhibits Aggregation-Induced Emission.
Polymers that exhibit aggregation-induced emission (AIE) find use, for example, as cell-imaging agents and as fluorometric sensors due to their unique optical properties. However, the structural diversity of AIE-active polymers has not necessarily advanced at the same rate as their applications. In this work, ring-opening metathesis polymerization is used to synthesize the first example of a polymer (Mn = 61,600 g mol-1 , Đ = 1.32) containing boron difluoride hydrazone (BODIHY) heterocycles in its repeating unit. The BODIHY monomer and polymer described absorb and emit in the visible region in solution (λabs = 428 and 429 nm, λem = 528 and 526 nm) and as thin films (λabs = 443 and 440 nm, λem = 535 and 534 nm). Monomer (ΦFilm = 10%) and polymer (ΦFilm = 6%) exhibit enhanced emission as thin films compared to solution (ΦSoln ≤ 1%) as well as AIE upon the addition of water to DMF solutions as a result of restriction of intramolecular motion. Enhancement factors for the monomer and polymer are determined to be 58 and 15, respectively. The title BODIHY polymer exhibited an earlier onset of AIE and enhanced sensitivity to solution viscosity when compared to the parent monomer.


Differences in CD4 cell counts at seroconversion and decline among 5739 HIV-1-infected individuals with well-estimated dates of seroconversion.
We studied repeated measurements of CD4 cell counts on 5739 HIV-1-infected individuals with reliably estimated dates of seroconversion (SC) aged > or =15 years at SC prior to initiation of highly active antiretroviral therapy (HAART) or AIDS using random effects models. Estimated CD4 cell count at SC differed significantly by sex, exposure group, and age, being higher in women, hemophilic men, and injection drug users (IDUs) as well as in those aged >40 years at SC. The rate of CD4 cell count decline did not differ significantly by sex; thus, differences between men and women were stable throughout the HIV-1 incubation period. There was a monotonic relationship between CD4 slopes and age at SC, with steeper slopes in older subjects. At 5 years after SC, the median difference in CD4 cell counts between the oldest (>40 years at SC) and youngest (16-20 years at SC) subjects was around 90 cells/microL. Mean rate of CD4 decline was significantly steeper in subjects diagnosed during acute infection. There was no evidence of a faster loss of CD4 cells in subjects who seroconverted after 1994. Apart from hemophilic men, who tended to have a steeper rate of CD4 decline on average, mean CD4 slopes did not differ by exposure category. These results suggest that before the initiation of HAART or other interventions based on immune status, consideration of demographic factors may be worthwhile.


Photocatalysts with internal electric fields.
The photocatalytic activity of materials for water splitting is limited by the recombination of photogenerated electron-hole pairs as well as the back-reaction of intermediate species. This review concentrates on the use of electric fields within catalyst particles to mitigate the effects of recombination and back-reaction and to increase photochemical reactivity. Internal electric fields in photocatalysts can arise from ferroelectric phenomena, p-n junctions, polar surface terminations, and polymorph junctions. The manipulation of internal fields through the creation of charged interfaces in hierarchically structured materials is a promising strategy for the design of improved photocatalysts.


Antifolate polyglutamylation and competitive drug displacement at dihydrofolate reductase as important elements in leucovorin rescue in L1210 cells.
Previous studies from this laboratory have shown that the addition of leucovorin to tumor cells dissociates methotrexate, but not methotrexate polyglutamates, from dihydrofolate reductase (L. H. Matherly, D. W. Fry, and I. D. Goldman, Cancer Res., 43: 2694-2699, 1983). To further assess the importance of these interactions to leucovorin rescue, antifolate growth inhibition toward L1210 cells in the presence of leucovorin was correlated with the metabolism of (6S)-5-formyl tetrahydrofolate to dihydrofolate as a measure of dihydrofolate reductase activity. Growth inhibition (greater than 95%) by methotrexate (5-10 microM) following its intracellular polyglutamylation during a 3-h preexposure, or by continuous treatment with high levels of the lipophilic antifolate, trimetrexate (1 microM), was only slightly diminished by 10 microM leucovorin (15-25%). High-pressure liquid chromatographic analyses of the derivatives formed from radiolabeled (6S)-5-formyl tetrahydrofolate under these conditions showed an incomplete conversion to dihydrofolate and metabolism to predominantly 10-formyl tetrahydrofolate. Neither of the antifolates interfered appreciably with the metabolism of the folate derivatives to polyglutamates. Growth inhibition in the presence of leucovorin correlated with the accumulation of dihydrofolate (1.5-2.2 nmol) from radiolabeled (6S)-5-formyl tetrahydrofolate, reflecting continued suppression of dihydrofolate reductase activity at these drug concentrations. With lower equitoxic levels of the trimetrexate (7.5 nM), the provision of leucovorin allowed for a restoration of cell growth to a level greater than 90% of control. Under these conditions, control levels of dihydrofolate (0.2 nmol) were formed from radiolabeled cofactor, consistent with sustained dihydrofolate reductase activity. These findings support a role for the activation of dihydrofolate reductase as an important component of the reversal of the effects of diaminoantifolates by leucovorin, presumably by a competitive displacement of drug from the enzyme. Since no displacement occurs in cells which have accumulated methotrexate polyglutamates, or in the presence of high levels of trimetrexate, it appears that the concentration of unbound drug within cells is a significant determinant of the extent of this competitive binding interaction. From these considerations, the high levels of methotrexate polyglutamates that accumulate in sensitive tumors relative to bone marrow and gastrointestinal cells would appear to represent an important factor for the selectivity of leucovorin rescue in vivo.


Influence of soil geochemical and physical properties on the sorption and bioaccessibility of chromium(III).
There are numerous Cr(III)-contaminated sites on Department of Defense (DoD) and Department of Energy (DOE) lands that are awaiting possible clean up and closure. Ingestion of contaminated soil by children is the risk driver that generally motivates the likelihood of site remediation. The purpose of this study was to develop a simple statistical model based on common soil properties to estimate the hioaccessibility of Cr(III)-contaminated soil upon ingestion. Thirty-five uncontaminated soils from seven major soil orders, whose properties were similar to numerous U.S. DoD contaminated sites, were treated with Cr(III) and aged. Statistical analysis revealed that Cr(III) sorption (e.g., adsorption and surface precipitation) by the soils was strongly correlated with the clay content, total inorganic C, pH, and the cation exchange capacity of the soils. Soils with higher quantities of clay, inorganic C (i.e., carbonates), higher pH, and higher cation exchange capacity generally sequestered more Cr(III). The amount of Cr(III) bioaccessible from the treated soils was determined with a physiologically based extraction test (PBET) that was designed to simulate the digestive process of the stomach. The bioaccessibility of Cr(III) varied widely as a function of soil type with most soils limiting bioaccessibility to <45 and <30% after I and 100 d soil-Cr aging, respectively. Statistical analysis showed the bioaccessibility of Cr(III) on soil was again related to the clay and total inorganic carbon (TIC) content of the soil. Bioaccessibility decreased as the soil TIC content increased and as the clay content decreased. The model yielded an equation based on common soil properties that could be used to predict the Cr(III) bioaccessibility in soils with a reasonable level of confidence.


The benefits of four weeks of melatonin treatment on circadian patterns in resistance-trained athletes.
Exercise can induce circadian phase shifts depending on the duration, intensity and frequency. These modifications are of special meaning in athletes during training and competition. Melatonin, which is produced by the pineal gland in a circadian manner, behaves as an endogenous rhythms synchronizer, and it is used as a supplement to promote resynchronization of altered circadian rhythms. In this study, we tested the effect of melatonin administration on the circadian system in athletes. Two groups of athletes were treated with 100 mg day(-1) of melatonin or placebo 30 min before bed for four weeks. Daily rhythm of salivary melatonin was measured before and after melatonin administration. Moreover, circadian variables, including wrist temperature (WT), motor activity and body position rhythmicity, were recorded during seven days before and seven days after melatonin or placebo treatment with the aid of specific sensors placed in the wrist and arm of each athlete. Before treatment, the athletes showed a phase-shift delay of the melatonin circadian rhythm, with an acrophase at 05:00 h. Exercise induced a phase advance of the melatonin rhythm, restoring its acrophase accordingly to the chronotype of the athletes. Melatonin, but not placebo treatment, changed daily waveforms of WT, activity and position. These changes included a one-hour phase advance in the WT rhythm before bedtime, with a longer nocturnal steady state and a smaller reduction when arising at morning than the placebo group. Melatonin, but not placebo, also reduced the nocturnal activity and the activity and position during lunch/nap time. Together, these data reflect the beneficial effect of melatonin to modulate the circadian components of the sleep-wake cycle, improving sleep efficiency.


Modulation of MutS ATP hydrolysis by DNA cofactors.
Escherichia coli MutS protein, which is required for mismatch repair, has a slow ATPase activity that obeys Michalelis-Menten kinetics. At 37 degrees C, the steady-state turnover rate for ATP hydrolysis is 1.0 +/- 0.3 min(-1) per monomer equivalent with a K(m) of 33 +/- 6 microM. Hydrolysis is competitively inhibited by the ATP analogues AMPPNP and ATPgammaS, with K(i) values of 4 microM in both cases, and by ADP with a K(i) of 40 microM. The rate of ATP hydrolysis is stimulated 2-5-fold by short hetero- and homoduplex DNAs. The concentration of DNA cofactor that yields half-maximal stimulation is lowest for oligodeoxynucleotide duplexes that contain a mismatched base pair. Pre-steady-state chemical quench analysis has demonstrated a substoichiometric initial burst of ADP formation by free MutS that is governed by a rate constant of 78 min(-1), indicating that the rate-limiting step for the steady-state reaction occurs after hydrolysis. Prebinding of MutS to homoduplex DNA does not alter the burst kinetics or amplitude but only increases the steady-state rate. In contrast, binding of the protein to heteroduplex DNA abolishes the burst of ADP formation, indicating that the rate-limiting step now occurs before hydrolysis. Gel filtration analysis indicates that the MutS dimer assembles into higher order oligomers in a concentration-dependent manner, and that ATP binding shifts this equilibrium to favor assembly. These results, together with kinetic findings, indicate nonequivalence of subunits within a MutS oligomer with respect to ATP hydrolysis and DNA binding.


The influence of sexually explicit online media on sex: do men who have sex with men believe they "do what they see"?
Over the past two decades, men who have sex with men (MSM) have engaged in increasing consumption of MSM-specific sexually explicit online media (i.e., online pornography). Furthermore, the amount of MSM-specific sexually explicit online media portraying unprotected anal intercourse (UAI) has increased, raising concerns about HIV transmission among the actors and the potential encouragement of risky sex among consumers. The influence of sexually explicit online media on sexual risk-taking, at present largely understudied, could lead to new avenues for innovative HIV-prevention strategies targeting at-risk MSM. In this preliminary assessment, in-depth qualitative interviews were conducted with 16 MSM in the Seattle area to elucidate MSM's perceptions about the influence of sexually explicit online media on their own and other MSM's sexual behaviors. Participants reported that sexually explicit online media: (1) plays an educational role, (2) increases comfort with sexuality, and (3) sets expectations about sexual behaviors. While participants overwhelmingly reported not feeling personally influenced by viewing UAI in sexually explicit online media, they believed viewing UAI increased sexual risk-taking among other MSM. Specifically, participants reported that the high prevalence of UAI in sexually explicit online media sends the message, at least to other MSM, that (1) engaging in UAI is common, (2) UAI is acceptable and "ok" to engage in, and (3) future partners will desire or expect UAI. Overall, this preliminary assessment indicates that sexually explicit online media exposure may have both positive (e.g., helping MSM become more comfortable with their sexuality) and negative (e.g., normalizing UAI) impacts on the sexual health of MSM and may be useful in the development of novel HIV-prevention interventions.


First Report of the Parasitic Plant Phelipanche aegyptiaca Infecting Kenaf in Israel.
Kenaf (Hibiscus cannabinus L.; Malvaceae) is an annual fiber crop that has recently been introduced as a niche crop in Israel, and grown mainly at areas with high summer temperatures. It is advantageous in crop rotation in these areas since it can rapidly accumulate biomass under high temperatures when other crops (e.g., tomato, sunflower, corn, and sorghum) cannot be cultivated. Additionally, the fact that it can be irrigated with waste water makes it attractive under these climatic conditions. Broomrapes (Phelipanche and Orobanche spp.) are chlorophyll-lacking obligatory root-parasitic plants that parasitize the root system of many field crops and vegetables (1). Parasitism by P. aegyptiaca has economic impacts on various crops belonging to several botanical families (e.g., Solanaceae, Apiaceae, Fabaceae, and Asteraceae). This parasitic weed is common in most agricultural areas in Israel including the coastal plain, Yisre'el Valley, the Jordan Valley, and the Negev Desert. High infection levels by P. aegyptiaca have result in a severe yield losses and quality reduction in these crops. Parasitism of P. aegyptiaca on kenaf was observed in September 2012 in a field located in Bet-Shean Valley (Latitude 32° 30' N; Longitude 35° 30' E; 105 m), with an average density of 0.7 plant/m2, in a total area of 0.3 ha. This crop was planted on May 2012 in plots that were previously affected. Infection did not lead to visible symptoms or damage to kenaf, but allowed seed production by the parasite. In order to verify that kenaf was a host of P. aegyptiaca, 10 samples of kenaf plants infected with P. aegyptiaca were taken to the lab and the root systems of the plant and the parasite were carefully washed. Cross-sections of the connection between kenaf and the parasite were taken and inspected for xylem connections under a compound microscope (BX61, Olympus) equipped with high-resolution digital camera (DP-70, Olympus), under 40× magnification. A clear xylem connection between the weed's tubercles and kenaf roots was observed, confirming the development of functional Phelipanche haustoria. To our knowledge, this is the first report of kenaf as a host for P. aegyptiaca. Kenaf has been reported to induce the germination of O. cernua in India (3). Cotton, another member of the Malvaceae, has also been reported to stimulate seed germination of O. minor (4) and P. aegyptiaca (2). However, to the best of our knowledge, this finding reports the first occurrence of a Malvaceae crop as a host for P. aegyptiaca. References: (1) H. Eizenberg et al. Weed Sci. 55:152, 2007. (2) M. Ghotbi et al. Int. J. Agri. Sci. 2:62, 2012. (3) G. V. G. Krishnamurthy et al. Indian J. Weed Sci. 9:95, 1977. (4) Y. Ma et al. Agron. J. 104:569, 2012.


A key commitment step in erythropoiesis is synchronized with the cell cycle clock through mutual inhibition between PU.1 and S-phase progression.
Hematopoietic progenitors undergo differentiation while navigating several cell division cycles, but it is unknown whether these two processes are coupled. We addressed this question by studying erythropoiesis in mouse fetal liver in vivo. We found that the initial upregulation of cell surface CD71 identifies developmentally matched erythroblasts that are tightly synchronized in S-phase. We show that DNA replication within this but not subsequent cycles is required for a differentiation switch comprising rapid and simultaneous committal transitions whose precise timing was previously unknown. These include the onset of erythropoietin dependence, activation of the erythroid master transcriptional regulator GATA-1, and a switch to an active chromatin conformation at the β-globin locus. Specifically, S-phase progression is required for the formation of DNase I hypersensitive sites and for DNA demethylation at this locus. Mechanistically, we show that S-phase progression during this key committal step is dependent on downregulation of the cyclin-dependent kinase p57(KIP2) and in turn causes the downregulation of PU.1, an antagonist of GATA-1 function. These findings therefore highlight a novel role for a cyclin-dependent kinase inhibitor in differentiation, distinct to their known function in cell cycle exit. Furthermore, we show that a novel, mutual inhibition between PU.1 expression and S-phase progression provides a "synchromesh" mechanism that "locks" the erythroid differentiation program to the cell cycle clock, ensuring precise coordination of critical differentiation events.


Transdermal power transfer for recharging implanted drug delivery devices via the refill port.
This paper describes a system for transferring power across a transdermal needle into a smart refill port for recharging implantable drug delivery systems. The device uses a modified 26 gauge (0.46 mm outer diameter) Huber needle with multiple conductive elements designed to couple with mechanical springs in the septum of the refill port of a drug delivery device to form an electrical connection that can sustain the current required to recharge a battery during a reservoir refill session. The needle is fabricated from stainless steel coated with Parylene, and the refill port septum is made from micromachined stainless steel contact springs and polydimethylsiloxane. The device properties were characterized with dry and wet ambient conditions. The needle and port pair had an average contact resistance of less than 2 Omega when mated in either environment. Electrical isolation between the system, the liquid in the needle lumen, and surrounding material has been demonstrated. The device was used to recharge a NiMH battery with currents up to 500 mA with less than 15 degrees C of resistive heating. The system was punctured 100 times to provide preliminary information with regard to device longevity, and exhibited about 1 Omega variation in contact resistance. The results suggest that this needle and refill port system can be used in an implant to enable battery recharging. This allows for smaller batteries to be used and ultimately increases the volume efficiency of an implantable drug delivery device.


Different reactions obtained using the same DNA detection reagents for Thai and Korean hepatopancreatic parvovirus of penaeid shrimp.
Hepatopancreatic parvovirus (HPV) can cause stunted growth and death in penaeid shrimp including Penaeus monodon. We used PCR primers and a commercial DNA probe designed from HPV of Penaeus chinensis (HPVchin) to examine HPV-infected Thai P. monodon (HPVmon). We found that the PCR primers produced a 732 bp DNA amplicon rather than the 350 bp amplicon obtained with HPVchin template and that the DNA probe gave weak to variable in situ DNA hybridization results. In addition, hybridization to PCR products from HPVmon was weak compared with hybridization with PCR products from HPVchin. By contrast, the 732 bp amplicon hybridized strongly with HPVmon-infected cells by in situ hybridization but not with uninfected shrimp tissue or other shrimp viruses, thus confirming its origin from HPVmon. Cloning, sequencing and analysis of the 732 bp amplicon showed that 696 bp (excluding the primer sequences) contained 47% GC content and had only 78% homology to 701 aligned bases from a 3350 bp DNA fragment of HPVchin from GenBank. These results explain why the reagents based on HPVchin gave a different PCR product and weak hybridization results with HPVmon, and they show that multiple primers or degenerate primers may be necessary for general detection of HPV varieties. Together with previously published information on the estimated total genome sizes for HPVchin (approximately 4 kb) and HPVmon (approximately 6 kb), these data support the contention that HPVchin and HPVmon are different varieties or species, in spite of their similar histopathology.


Effects of weight loss and long-term weight maintenance with diets varying in protein and glycemic index on cardiovascular risk factors: the diet, obesity, and genes (DiOGenes) study: a randomized, controlled trial.
We sought to separately examine the effects of either weight loss or diets varying in protein content and glycemic index without further changes in body weight on cardiovascular risk factors within the Diet, Obesity, and Genes study (DiOGenes). DiOGenes is a pan-European controlled dietary intervention study in 932 overweight adults who first lost body weight on an 8-week low-calorie diet and were then randomized to 1 of 5 ad libitum diets for 26 weeks. The diets were either high or low protein or high or low glycemic index in 4 combinations or control. Weight loss (-11.23 kg; 95% confidence interval, -11.54 to -10.92; P<0.001) reduced high-sensitivity C-reactive protein (-1.15 mg/L; 95% confidence interval, -1.30 to -0.41; P<0.001), low- and high-density lipoprotein cholesterol, triglycerides, and blood pressure. During the 26-week weight maintenance period in the intention-to-treat analysis, the further decrease of high-sensitivity C-reactive protein blood levels was -0.46 mg/L greater (95% confidence interval, -0.79 to -0.13) in the groups assigned to low-glycemic-index diets than in those on high-glycemic-index diets (P<0.001). Groups on low-protein diets achieved a -0.25 mg/L greater reduction in high-sensitivity C-reactive protein (95% confidence interval, -0.59 to -0.17) than those on high-protein diets (P<0.001), whereas lipid profiles and blood pressure were not differently affected. This large-scale intervention study clearly separates weight loss from dietary composition-related effects. Low-glycemic-index carbohydrates and, to a lesser extent, low-protein intake may specifically reduce low-grade inflammation and associated comorbidities in overweight/obese adults. http://www.clinicaltrials.gov. Unique identifier: NCT00390637.


Unusual haplotypic structure of IL8, a susceptibility locus for a common respiratory virus.
Interleukin-8 (IL8) is believed to play a role in the pathogenesis of bronchiolitis, a common viral disease of infancy, and a recent U.K. family study identified an association between this disease and the IL8-251A allele. In the present study we report data, from a different set of families, which replicate this finding; combined analysis of 194 nuclear families through use of the transmission/disequilibrium test gives P = .001. To explore the underlying genetic cause, we identified nine single-nucleotide polymorphisms (SNPs) in a 7.6-kb segment spanning the IL8 gene and its promoter region and used six of these SNPs to define the haplotypic structure of the IL8 locus. The IL8-251A allele resides on two haplotypes, only one of which is associated with disease, suggesting that this may not be the functional allele. Europeans show an unusual haplotype genealogy that is dominated by two common haplotypes differing at multiple sites, whereas Africans have much greater haplotypic diversity. These marked haplotype-frequency differences give an F(ST) of.25, and, in the European sample, both Tajima's D statistic (D = 2.58, P = .007) and the Hudson/Kreitman/Aguade test (chi(2) = 4.9, P = .03) reject neutral equilibrium, suggesting that selective pressure may have acted on this locus.


The Effect of a Mechanical Arm System on Portable Grinder Vibration Emissions.
Mechanical arm systems are commonly used to support powered hand tools to alleviate ergonomic stressors related to the development of workplace musculoskeletal disorders. However, the use of these systems can increase exposure times to other potentially harmful agents such as hand-transmitted vibration. To examine how these tool support systems affect tool vibration, the primary objectives of this study were to characterize the vibration emissions of typical portable pneumatic grinders used for surface grinding with and without a mechanical arm support system at a workplace and to estimate the potential risk of the increased vibration exposure time afforded by the use of these mechanical arm systems. This study also developed a laboratory-based simulated grinding task based on the ISO 28927-1 (2009) standard for assessing grinder vibrations; the simulated grinding vibrations were compared with those measured during actual workplace grinder operations. The results of this study demonstrate that use of the mechanical arm may provide a health benefit by reducing the forces required to lift and maneuver the tools and by decreasing hand-transmitted vibration exposure. However, the arm does not substantially change the basic characteristics of grinder vibration spectra. The mechanical arm reduced the average frequency-weighted acceleration by about 24% in the workplace and by about 7% in the laboratory. Because use of the mechanical arm system can increase daily time-on-task by 50% or more, the use of such systems may actually increase daily time-weighted hand-transmitted vibration exposures in some cases. The laboratory acceleration measurements were substantially lower than the workplace measurements, and the laboratory tool rankings based on acceleration were considerably different than those from the workplace. Thus, it is doubtful that ISO 28927-1 is useful for estimating workplace grinder vibration exposures or for predicting workplace grinder acceleration rank orders.


How does monetary loss empathy modulate generosity in economic sharing behavior? An ERPs study.
Previous studies have shown that generosity is driven by empathy and that both generosity in economic sharing behavior and monetary loss empathy decay as the social distance increases. However, it is still unclear whether this decay in economic sharing generosity can be influenced by the decay in monetary loss empathy. In the current study, we carried out two experiments to investigate this issue to deepen our understanding of the relationship between monetary loss empathy and generosity in economic sharing behavior. Our results show that in the observation group (observers watch their friend, and a stranger plays a gambling game), a negative correlation between log-transformed k value (ln(k)) and the distinction of d-FRN (feedback-related negativity difference between gain and loss) between friends and strangers was observed. However, in the execution group (executors play a gambling game themselves and watch a stranger play the same gambling game), there was no significant correlation between ln(k) and the distinction of d-FRN between self and strangers. Current results indicate that the decayed generosity across different social distances in economic sharing behavior can be modulated by the decayed monetary loss empathy. The study adds weight to the relationship between decayed monetary loss empathy and decayed generosity in sharing economic behavior at the level of social distance and provides electrophysiological evidence.


Identification of teleost Thy-1 and association with the microdomain/lipid raft reggie proteins in regenerating CNS axons.
During regeneration, retinal ganglion cell axons in fish upregulate a cell surface protein that is recognized by the monoclonal antibody (mAB) M802. M802 antigen appeared to be linked to the intracellular, membrane-associated lipid raft/microdomain proteins reggie-1 and reggie-2 that were previously shown to be reexpressed in axon-regenerating neurons [Development 124 (1997), 577]. Here, we report the isolation of the M802 antigen and its identification as the teleost homolog of mammalian Thy-1. Fish Thy-1 is detected in the same detergent-insoluble lipid raft fractions from a fibroblast cell line and from axon regenerating retinae as reggie-1 and 2. Importantly, mAB M802 coimmunoprecipitates reggie-1 and 2 from this lipid raft fraction, implying that fish Thy-1 and reggies interact. This correlates with their colocalization in growing cell processes after M802 antigen/Thy-1 activation with mAB M802. These findings suggest a role of clustered M802 antigen/Thy-1 in reggie raft microdomains for cell growth and axon regeneration.


Time course of synergistic interaction between DOCA and salt on blood pressure: roles of vasopressin and hepatic osmoreceptors.
In DOCA-salt rats, the time course of the synergistic interaction between osmolality and DOCA to produce hypertension is unknown. Therefore, in rats 2 wk after implantation of subcutaneous silicone pellets containing DOCA (65 mg) or no drug (sham), we determined blood pressure (BP) and heart rate (HR) responses, using telemetric pressure transducers, during 2 wk of excess salt ingestion (1% NaCl in drinking water). BP was unaltered in sham rats after increased salt, but in DOCA rats BP increased within 4 h. The initial hypertension of 30-35 mmHg stabilized within 2 days, followed approximately 5 days later by a further increment of approximately 30 mmHg. HR first decreased during the dark phase; the second phase was linked to an abrupt increase in HR and BP variability and decreased HR variability. Pressor responses to acute intravenous hypertonic saline infusion were doubled in DOCA-treated rats via vasopressin and nonvasopressin mechanisms. Only in DOCA-treated rats, portal vein hypertonic saline infusion increased BP, which was prevented by V(1) vasopressin blockade. After 2 wk of DOCA-salt, oral ingestion of water rapidly decreased BP. Intraportal infusion of water did not lower BP in DOCA-salt rats, suggesting that hepatic osmoreceptors were not involved. In summary, the hypertension of DOCA-treated rats consuming excess salt exhibits multiple phases and can be rapidly reversed. Hypertonicity-induced vasopressin and nonvasopressin pressor mechanisms that are augmented by DOCA, and hepatic osmoreceptors may contribute to the initial developmental phase. With time, combined DOCA-salt induces marked changes in the regulation of the autonomic nervous system, which may favor hypertension development.


"Neither we are satisfied nor they"-users and provider's perspective: a qualitative study of maternity care in secondary level public health facilities, Uttar Pradesh, India.
Quality of care provided during childbirth is a critical determinant of preventing maternal mortality and morbidity. In the studies available, quality has been assessed either from the users' perspective or the providers'. The current study tries to bring both perspectives together to identify common key focus areas for quality improvement. This study aims to assess the users' (recently delivered women) and care providers' perceptions of care to understand the common challenges affecting provision of quality maternity care in public health facilities in India. A qualitative design comprising of in-depth interviews of 24 recently delivered women from secondary care facilities and 16 health care providers in Uttar Pradesh, India. The data were analysed thematically to assess users' and providers' perspectives on the common themes. The common challenges experienced regarding provision of care were inadequate physical infrastructure, irregular supply of water, electricity, shortage of medicines, supplies, and gynaecologist and anaesthetist to manage complications, difficulty in maintaining privacy and lack of skill for post-delivery counselling. However, physical access, cleanliness, interpersonal behaviour, information sharing and out-of-pocket expenditure were concerns for only users. Similarly, providers raised poor management of referral cases, shortage of staff, non-functioning of blood bank, lack of incentives for work as their concerns. The study identified the common themes of care from both the perspectives, which have been foundrelevant in terms of challenges identified in many developing countries including India. The study framework identified new themes like management of emergencies in complicated cases, privacy and cost of care which both the group felt is relevant in the context of providing quality care during childbirth in low resource setting. The key challenges identified by both the groups can be prioritized, when developing quality improvement program in the health facilities. The identified components of care can match the supply with the demand for care and make the services truly responsive to user needs. The study highlights infrastructure, human resources, supplies and medicine as priority areas of quality improvement in the facility as perceived by both users and providers, nevertheless the interpersonal aspect of care primarily reported by the users must also not be ignored.


Copy Number Variation of Mitochondrial DNA Genes in Pneumocystis jirovecii According to the Fungal Load in BAL Specimens.
Pneumocystis jirovecii is an unculturable fungus and the causative agent of Pneumocystis pneumonia, a life-threatening opportunistic infection. Although molecular diagnosis is often based on the detection of mtLSU rRNA mitochondrial gene, the number of copies of mitochondrial genes had not been investigated. We developed and optimized six real-time PCR assays in order to determine the copy number of four mitochondrial genes (mtSSU rRNA, mtLSU rRNA, NAD1, and CYTB) in comparison to nuclear genome (DHPS and HSP70) and tested 84 bronchoalveolar fluids of patients at different stages of the infection. Unexpectedly, we found that copy number of mitochondrial genes varied from gene to gene with mtSSU rRNA gene being more represented (37 copies) than NAD1 (23 copies), mtLSU rRNA (15 copies) and CYTB (6 copies) genes compared to nuclear genome. Hierarchical clustering analysis (HCA) allowed us to define five major clusters, significantly associated with fungal load (p = 0.029), in which copy number of mitochondrial genes was significantly different among them. More importantly, copy number of mtLSU rRNA, NAD1, and CYTB but not mtSSU rRNA differed according to P. jirovecii physiological state with a decreased number of copies when the fungal load is low. This suggests the existence of a mixture of various subspecies of mtDNA that can harbor different amplification rates. Overall, we revealed here an unexpected variability of P. jirovecii mtDNA copy number that fluctuates according to P. jirovecii's physiological state, except for mtSSU that is the most stable and the most present mitochondrial gene.


The modular organization of brain cortical connectivity across the human lifespan.
The network architecture of the human brain contributes in shaping neural activity, influencing cognitive and behavioral processes. The availability of neuroimaging data across the lifespan allows us to monitor how this architecture reorganizes, influenced by processes like learning, adaptation, maturation, and senescence. Changing patterns in brain connectivity can be analyzed with the tools of network science, which can be used to reveal organizational principles such as modular network topology. The identification of network modules is fundamental, as they parse the brain into coherent sub-systems and allow for both functional integration and segregation among different brain areas. In this work we examined the brain's modular organization by developing an ensemble-based multilayer network approach, allowing us to link changes of structural connectivity patterns to development and aging. We show that modular structure exhibits both linear and nonlinear age-related trends. In the early and late lifespan, communities are more modular, and we track the origins of this high modularity to two different substrates in brain connectivity, linked to the number and the weights of the intra-clusters edges. We also demonstrate that aging leads to a progressive and increasing reconfiguration of modules and a redistribution across hemispheres. Finally, we identify those brain regions that most contribute to network reconfiguration and those that remain more stable across the lifespan.


The effect of backbone flexibility on site-selective modification of macrocycles.
As the emerging modality in drug discovery, macrocycles represent topical targets of chemical synthesis. However, the influence of conformational effects on their reactivity has not received enough attention. We demonstrate that the regiochemistry of nucleophilic attack on aziridine-containing macrocyclic systems varies according to macrocycle ring size and that the influence of remote torsional interactions in the macrocycle ultimately determines the regioselectivity of the reactions. These findings should facilitate the development of divergent strategies of macrocycle synthesis using post-cyclization modification.


An MVA vaccine overcomes tolerance to human p53 in mice and humans.
The cellular regulatory protein p53 is overexpressed by almost 50% of all malignancies making it an attractive target for a vaccine approach to cancer. A number of immunotherapy approaches targeting p53 have been evaluated successfully in murine models, but translation of these preclinical findings to the clinic has been unsuccessful. Prior studies in our laboratory employing murine models demonstrated that a modified vaccinia virus Ankara (MVA) vaccine expressing murine p53 could stimulate p53 specific immunity. Systemic administration of the MVA vaccine was able to effect the rejection of established tumors. To better understand the immunologic mechanisms that underlie the vaccine function of human p53, we utilized a murine model in which the murine germ line copy of p53 was replaced with a modified human one. These mice, referred to as Hupki, were evaluated as a tolerant model to explore the capacity of MVA expressing human p53 to overcome tolerance and reject human p53-expressing tumors. MVAp53 immunization of Hupki mice resulted in the generation of p53-specific CD8(+) T cells and the rejection of a highly aggressive murine mammary carcinoma cell line 4T1(H-2d) transfected with human p53 (4T1p53). An immunologic correlate of tumor protection was evaluated utilizing an overlapping peptide library spanning the full length of human p53. This reagent was also used in combination with MVAp53 to stimulate p53-specific CD8(+) T cell responses in cancer patients. These studies demonstrate the potential of MVAp53 to overcome tolerance to p53 for cancer immunotherapy.


Dietary factors in Parkinson's disease: the role of food groups and specific foods.
The association between self-reported past food intake and Parkinson's disease (PD) was investigated in a case-control study of men and women aged 40-89 years. Newly diagnosed idiopathic PD cases were ascertained from neurologists, and from outpatient and pharmacy computerized databases, at the Group Health Cooperative (GHC) clinics in the Puget Sound region of Washington state. Control subjects were chosen from the GHC patient roster and had no reported history of diagnosed neurodegenerative disease. Dietary data were obtained from structured questionnaires. An increase in PD risk with increasing intake was noted for foods that contain animal fat and foods containing vitamin D. Intake of fruits, vegetables, meats, bread and cereals, or foods containing vitamins A, C, E, or iron was not significantly related to PD risk. Vitamin use, in general, was also not found to be related to PD risk, although a significant trend of increasing risk of PD was noted for intake of vitamin A supplements. Although these data support previous findings of no association of past intake with most food groups and PD risk, they confirm an increased risk of PD associated with foods containing animal fat.


Acute toxoplasmosis leads to lethal overproduction of Th1 cytokines.
Virulence in Toxoplasma gondii is strongly influenced by the genotype of the parasite. Type I strains uniformly cause rapid death in mice regardless of the host genotype or the challenge dose. In contrast, the outcome of infections with type II strains is highly dependent on the challenge dose and the genotype of the host. To understand the basis of acute virulence in toxoplasmosis, we compared low and high doses of the RH strain (type I) and the ME49/PTG strain (type II) of T. gondii in outbred mice. Differences in virulence were reflected in only modestly different growth rates in vivo, and both strains disseminated widely to different tissues. The key difference in the virulent RH strain was the ability to reach high tissue burdens rapidly following a low dose challenge. Lethal infections caused by type I (RH) or type II (PTG) strain infections were accompanied by extremely elevated levels of Th1 cytokines in the serum, including IFN-gamma, TNF-alpha, IL-12, and IL-18. Extensive liver damage and lymphoid degeneration accompanied the elevated levels of cytokines produced during lethal infection. Increased time of survival following lethal infection with the RH strain was provided by neutralization of IL-18, but not TNF-alpha or IFN-gamma. Nonlethal infections with a low dose of type II PTG strain parasites were characterized by a modest induction of Th1 cytokines that led to control of infection and minimal damage to host tissues. Our findings establish that overstimulation of immune responses that are normally necessary for protection is an important feature of acute toxoplasmosis.


Aquatic photochemistry of sulfamethazine: multivariate effects of main water constituents and mechanisms.
The ubiquity of sulfonamides (SAs) in natural waters requires insight into their environmental fate for ecological risk assessment. Extensive studies focused on the effect of univariate water constituents on the photochemical fate of SAs, yet the multivariate effects of water constituents in environmentally relevant concentrations on SA photodegradation are poorly understood. Here, response surface methodology was employed to explore the integrative effects of main water constituents (dissolved organic matter (DOM), NO3-, HCO3-, Cu2+) on the photodegradation of a representative SA (sulfamethazine). Results showed that besides single factors, interaction of factors also significantly impacted the photodegradation. Radical scavenging experiments indicated that triplet-excited DOM (3DOM*) was responsible for the enhancing effect of DOM on the photodegradation. Additionally, DOM may also quench the 3DOM*-mediated oxidation intermediate of sulfamethazine causing the inhibiting effect of DOM-DOM interaction. We also found that HCO3- was oxidized by triplet-excited sulfamethazine producing CO3˙-, and the high reactivity of CO3˙- with sulfamethazine (second-order rate constant 2.2 × 108 M-1 s-1) determined by laser flash photolysis revealed the enhancing photodegradation mechanism of HCO3-. This study is among the first attempts to probe the photodegradation of SAs considering the integrative effects of water constituents, which is important in accurate ecological risk assessment of organic pollutants in the aquatic environment.


Cardiometabolic risk in polycystic ovary syndrome.
Polycystic ovary syndrome (PCOS) is a common disorder in women of reproductive age. Besides hyperandrogenism, oligomenorrhea and fertility issues, it is associated with a high prevalence of metabolic disorders and cardiovascular risk factors. Several genetic polymorphisms have been identified for possible associations with cardiometabolic derangements in PCOS. Different PCOS phenotypes differ significantly in their cardiometabolic risk, which worsens with severity of androgen excess. Due to methodological difficulties, longer time-scale data about cardiovascular morbidity and mortality in PCOS and about possible beneficial effects of different treatment interventions is missing leaving many issues regarding cardiovascular risk unresolved.


Anterior minimally invasive approach for total hip replacement: five-year survivorship and learning curve.
Opponents associate minimally invasive total hip replacement (THR) with additional risks, potentially resulting in increased implant failure rates. The purpose was to document complications, quality of implant positioning and five-year survivorship of THR using the AMIS approach and to test the hypothesis that eventual high complication and revision rates would be limited to an early series and be avoided by junior surgeons who get trained by a senior surgeon. A consecutive series of 150 primary THR implanted during the introduction of the AMIS technique in the department was retrospectively analysed for complications, implant positioning and implant survival after a minimum of five years. Survivorship curves of implants were compared between different surgeons and time periods. Due to implant revision for any reason the five-year survival rate was 94.6%, 78.9% for the first 20 and 96.8% for the following 130 AMIS procedures (p = 0.001). The hazard ratio for implant failure was 0.979 indicating a risk reduction of 2% every further case. The five-year implant survivorship of those procedures performed by two junior surgeons was 97.7%. We conclude that adoption of AMIS temporarily exposed patients to a higher risk of implant revisions, which normalised after the first 20 cases and that experience from a single surgeon's learning curve could effectively be taught to junior surgeons.


Fluoride intake and cortical and trabecular bone characteristics in adolescents at age 17: A prospective cohort study.
To investigate the associations between period-specific and cumulative fluoride (F) intakes from birth to age 17 years, and radial and tibial bone measures obtained using peripheral quantitative computed tomography (pQCT). Participants (n = 380) were recruited from hospitals at birth and continued their participation in the ongoing Iowa Fluoride Study/Iowa Bone Development Study until age 17. Fluoride intakes from water, other beverages, selected foods, dietary fluoride supplements and dentifrice were determined every 1.5-6 months using detailed questionnaires. Associations between F intake and bone measures (cortical and trabecular bone mineral content [BMC], density and strength) were determined in bivariate and multivariable analyses adjusted for height, weight, maturity offset, physical activity, and daily calcium and protein intake using robust regression analysis. Fluoride intake ranged from 0.7 to 0.8 mg F/d for females and from 0.7 to 0.9 mg F/d for males. Spearman correlations between daily F intake and pQCT bone measures were weak. For females, Spearman correlations ranged from r = -.08 to .21, and for males, they ranged from r = -.03 to .30. In sex-specific, height-, weight- and maturity offset- partially adjusted regression analyses, associations between females' fluoride intake and bone characteristics were almost all negative; associations for males were mostly positive. In the fully adjusted models, which also included physical activity, and protein and calcium intakes, no significant associations were detected for females; significant positive associations were detected between F intake from 14 to 17 years and tibial cortical bone content (β = 21.40, P < .01) and torsion strength (β = 175.06, P < .01) for males. In this cohort of 17-year-old adolescents, mostly living in optimally fluoridated areas, lifelong F intake from combined sources was weakly associated with bone pQCT measures.


Design, mechanism of action, bioavailability and therapeutic effects of mn porphyrin-based redox modulators.
Based on aqueous redox chemistry and simple in vivo models of oxidative stress, Escherichia coli and Saccharomyces cerevisiae, the cationic Mn(III) N-substituted pyridylporphyrins (MnPs) have been identified as the most potent cellular redox modulators within the porphyrin class of drugs; their efficacy in animal models of diseases that have oxidative stress in common is based on their high ability to catalytically remove superoxide, peroxynitrite, carbonate anion radical, hypochlorite, nitric oxide, lipid peroxyl and alkoxyl radicals, thus suppressing the primary oxidative event. While doing so MnPs could couple with cellular reductants and redox-active proteins. Reactive species are widely accepted as regulators of cellular transcriptional activity: minute, nanomolar levels are essential for normal cell function, while submicromolar or micromolar levels impose oxidative stress, which is evidenced in increased inflammatory and immune responses. By removing reactive species, MnPs affect redox-based cellular transcriptional activity and consequently secondary oxidative stress, and in turn inflammatory processes. The equal ability to reduce and oxidize superoxide during the dismutation process and recently accumulated results suggest that pro-oxidative actions of MnPs may also contribute to their therapeutic effects. All our data identify the superoxide dismutase-like activity, estimated by log k(cat)O2-*), as a good measure for the therapeutic efficacy of MnPs. Their accumulation in mitochondria and their ability to cross the blood-brain barrier contribute to their remarkable efficacy. We summarize herein the therapeutic effects of MnPs in cancer, central nervous system injuries, diabetes, their radioprotective action and potential for imaging. Few of the most potent modulators of cellular redox-based pathways, MnTE2-PyP5+, MnTDE-2-ImP5+, MnTnHex-2-PyP5+ and MnTnBuOE-2-PyP5+, are under preclinical and clinical development.


Prenatal methylazoxymethanol exposure alters evoked responses in fetal rats.
Although there is considerable interest in identifying methods to detect central nervous system impairment early in development, few behavioral assessment tools are available for detecting CNS deficits in the fetus. In the present study, methylazoxymethanol [MAM; Midwest Research Institute, (MRI)] was used to induce deficits in CNS development in fetal rats to assess effects on coordinated fetal behavior. Fetuses were exposed by administering MAM to pregnant rats on E17 of gestation via intraperitoneal injection and then were prepared for behavioral testing 3 days later on E20. After externalization from the uterus into a warm saline bath, fetal subjects received either an intraoral infusion of lemon extract to evoke a facial wiping response or were presented with an artificial nipple to evoke an oral grasping response. Interlimb coordination and paw-face contact during facial wiping were disrupted in MAM-exposed fetuses. Similarly, MAM exposure diminished the ability of fetuses to grasp or maintain oral contact with the artificial nipple. Although clear disruptions of movement coordination were seen in the MAM-treated subjects, there were no significant differences from saline controls in weight or anatomical measures. Together, these findings suggest that behavioral assessments of fetal motor coordination may be useful in identifying neural insult during prenatal development.


Dysregulation of apoptosis is a major mechanism in the lymph node involvement in colorectal carcinoma.
The purpose of this study was to define gene expression profile changes in colorectal tumors in order to identify target genes involved in neoplastic progression. cDNA microarray analysis was used to detect differences in gene expression profiles between colon tumor samples obtained from 20 patients in different tumor stages. Genes included in the cDNA microarray were selected according to their role in the cell cycle, apoptosis process, drug resistance and transcription factor regulation. Cluster analysis showed 2 well differentiated gene expression profiles between colorectal tumors with or without lymph node involvement. Some of these genes are important regulators of apoptotic pathways (DAD1, APO3, DRAK1 or BIK), suggesting that this process could be associated with node involvement. Subsequent analysis of certain genes identified in the microarray analysis were confirmed by quantitative real-time PCR. Our data suggest that microarray technology could discriminate between the involvement of regional lymph node in colon cancer where apoptosis-related genes would be implied. This preliminary analysis also suggests that the gene expression profile may be useful in improving risk-group stratification.


Huntingtin promotes mTORC1 signaling in the pathogenesis of Huntington's disease.
In patients with Huntington's disease (HD), the protein huntingtin (Htt) has an expanded polyglutamine (poly-Q) tract. HD results in early loss of medium spiny neurons in the striatum, which impairs motor and cognitive functions. Identifying the physiological role and molecular functions of Htt may yield insight into HD pathogenesis. We found that Htt promotes signaling by mTORC1 [mechanistic target of rapamycin (mTOR) complex 1] and that this signaling is potentiated by poly-Q-expanded Htt. Knocking out Htt in mouse embryonic stem cells or human embryonic kidney cells attenuated amino acid-induced mTORC1 activity, whereas overexpressing wild-type or poly-Q-expanded Htt in striatal neuronal cells increased basal mTOR activity. Striatal cells expressing endogenous poly-Q-expanded Htt showed an increase in the number and size of mTOR puncta on the perinuclear regions compared to cells expressing wild-type Htt. Pull-down experiments indicated that amino acids stimulated the interaction of Htt and the guanosine triphosphatase (GTPase) Rheb (a protein that stimulates mTOR activity), and that Htt forms a ternary complex with Rheb and mTOR. Pharmacologically inhibiting PI3K (phosphatidylinositol 3-kinase) or knocking down Rheb abrogated mTORC1 activity induced by expression of a poly-Q-expanded amino-terminal Htt fragment. Moreover, striatum-specific deletion of TSC1, encoding tuberous sclerosis 1, a negative regulator of mTORC1, accelerated the onset of motor coordination abnormalities and caused premature death in an HD mouse model. Together, our findings demonstrate that mutant Htt contributes to the pathogenesis of HD by enhancing mTORC1 activity.


Broadly neutralizing antibodies elicited by the hypervariable neutralizing determinant of HIV-1.
The principal neutralizing determinant (PND) of human immunodeficiency virus (HIV)-1 resides within the V3 loop of the envelope protein. Antibodies elicited by peptides of this region were able to neutralize diverse isolates. Serum from one of three animals immunized with the human T cell lymphoma virus (HTLV)-IIIMN PND peptide, RP142, neutralized MN and the sequence-divergent HTLV-IIIB isolate. Serum from one of three animals immunized with a 13-amino acid IIIB PND peptide (RP337) also neutralized both of these isolates. Characterization of these sera revealed that the cross-neutralizing antibodies bound the amino acid sequence GlyProGlyArgAlaPhe (GPGRAF) that is present in both isolates. This sequence is frequently found in the PNDs analyzed in randomly selected HIV-1 isolates. Sera from two rabbits immunized with a peptide containing only the GPGRAF residues neutralized divergent isolates, including IIIB and MN.


The Effects of a Sport-Specific Maximal Strength and Conditioning Training on Critical Velocity, Anaerobic Running Distance, and 5-km Race Performance.
To investigate the effects of a sport-specific maximal 6-wk strength and conditioning program on critical velocity (CV), anaerobic running distance (ARD), and 5-km time-trial performance (TT). 16 moderately trained recreational endurance runners were tested for CV, ARD, and TT performances on 3 separate occasions (baseline, midstudy, and poststudy). Participants were randomly allocated into a strength and conditioning group (S&C; n = 8) and a comparison endurance-training-only group (EO; n = 8). During the first phase of the study (6 wk), the S&C group performed concurrent maximal strength and endurance training, while the EO group performed endurance-only training. After the retest of all variables (midstudy), both groups subsequently, during phase 2, performed another 6 wk of endurance-only training that was followed by poststudy tests. No significant change for CV was identified in either group. The S&C group demonstrated a significant decrease for ARD values after phases 1 and 2 of the study. TT performances were significantly different in the S&C group after the intervention, with a performance improvement of 3.62%. This performance increase returned close to baseline after the 6-wk endurance-only training. Combining a 6-wk resistance-training program with endurance training significantly improves 5-km TT performance. Removing strength training results in some loss of those performance improvements.


Expression and segregation of HL-A antigens in D98/AH-2 by lymphocyte and fibroblast hybrids.
Somatic cell hybrids between D98/AH-2 and fibroblasts or peripheral blood lymphocytes were analysed for the expression of HL-A antigens. The hybrids express the antigens of the donor lymphocytes or fibroblasts as well as other HL-A specificities from D98/AH-2, a HeLa derivative on which no HL-A could be detected by direct cytotoxicity. Segregant hybrid clones could be used to determine HL-A haplotypes and define the linkage relationships between these haplotypes and the PGM3 alleles expressed in the hybrids. An analysis of the HL-A antigens on D98/AH-2 and other HeLa derivates was done using absorption and a modified cytotoxicity assay.


Idiopathic thrombocytopenic purpura (ITP) - new era for an old disease.
Immune thrombocytopenia is an autoimmune hematological disorder characterized by severely decreased platelet count of peripheral cause: platelet destruction via antiplatelet antibodies which may also affect marrow megakaryocytes. Patients may present in critical situations, with cutaneous and/or mucous bleeding and possibly life-threatening organ hemorrhages (cerebral, digestive, etc.) Therefore, rapid diagnosis and therapeutic intervention are mandatory. Corticotherapy represents the first treatment option, but as in any autoimmune disorder, there is a high risk of relapse. Second line therapy options include: intravenous immunoglobulins, thrombopoietin receptor agonists, rituximab or immunosuppression, but their benefit is usually temporary. Moreover, the disease generally affects young people who need repeated and prolonged treatment and hospitalization and therefore, it is preferred to choose a long term effect therapy. Splenectomy - removal of the site of platelet destruction - represents an effective and stable treatment, with 70-80% response rate and low complications incidence. A challenging situation is the association of ITP with pregnancy, which further increases the risk due to the immunodeficiency of pregnancy, major dangers of bleeding, vital risks for mother and fetus, potential risks of medication, necessity of prompt intervention in the setting of specific obstetrical situations - delivery, pregnancy loss, obstetrical complications, etc. We present an updated review of the current clinical and laboratory data, as well as a detailed analysis of the available therapeutic options with their benefits and risks, and also particular associations (pregnancy, relapsed and refractory disease, emergency treatment).


Improving sleep and cognition by hypnotic suggestion in the elderly.
Sleep quality markedly declines across the human lifespan. Particularly the amount of slow-wave sleep (SWS) decreases with age and this decrease is paralleled by a loss of cognitive functioning in the elderly. Here we show in healthy elderly females that the amount of SWS can be extended by a hypnotic suggestion "to sleep deeper" before sleep. In a placebo-controlled cross-over design, participants listened to hypnotic suggestions or a control tape before a midday nap while high density electroencephalography was recorded. After the hypnotic suggestion, we observed a 57% increase in SWS in females suggestible to hypnosis as compared to the control condition. Furthermore, left frontal slow-wave activity (SWA), characteristic for SWS, was significantly increased, followed by a significant improvement in prefrontal cognitive functioning after sleep. Our results suggest that hypnotic suggestions might be a successful alternative for widely-used sleep-enhancing medication to extend SWS and improve cognition in the elderly.


Maximum blood flow and microvascular regulatory responses in systemic sclerosis.
Objectives. To investigate microvascular function using laser Doppler imaging (LDI) following response to hyperaemia, neurovascular reflex and iontophoresis in systemic sclerosis (SSc) in comparison with primary Raynaud's phenomenon (PRP) and age-matched controls. A secondary aim was to evaluate if SSc patients with a higher Medsger vascular score have lower endothelial responses. Methods. Twenty patients with SSc, 10 PRP and 17 controls were studied. Patients with SSc were scored using the vascular component of the Medsger severity scale. A baseline LDI scan was performed on the dorsal aspect of both hands. Digital responses were quantified following maximum hyperaemic response (MHR), contralateral vasoconstrictor response (CLVc) and iontophoresis with acetylcholine (Ach)-endothelial dependent and sodium nitroprusside (SNP)-endothelial independent. Mean blood flow was quantified over a standard region of interest. Results. MHR was lower in SSc patients compared with controls (P < 0.001). A similar trend was seen when comparing SSc with PRP although this did not reach significance (P = 0.07). CLVc and Ach/MHR were lower in SSc vs PRP (P < 0.05) and controls (P < 0.001). No difference was observed in MHR, CLVc and Ach/MHR between PRP and controls. Overall, SNP/MHR was similar in all the three groups. SSc patients with a higher Medsger vascular score had lower endothelial-dependent (P < 0.01) and independent (P < 0.05) responses. Conclusion. SSc patients have abnormal microvascular regulatory responses compared with PRP and controls. This study also suggests that the degree of endothelial dysfunction may be related to the degree of peripheral vascular involvement.


Histological heterogeneity of Ewing's sarcoma/PNET: an immunohistochemical analysis of 415 genetically confirmed cases with clinical support.
Ewing's sarcoma (ES)/peripheral neuroectodermal tumor (PNET) are malignant neoplasms affecting children and young adults. We performed a study to typify the histological diversity and evaluate antibodies that may offer diagnostic/prognostic support. In total, 415 cases of genetically confirmed paraffin-embedded ES/PNET were analyzed on whole sections and in tissue microarrays. This study confirms the structural heterogeneity of ES/PNET, distinguishing three major subtypes: conventional ES (280 cases); PNET (53 cases); and atypical ES/PNET (80), including large cells, vascular-like patterns, spindle pattern, and adamantinoma-like configuration. All cases presented positivity for at least three of the four tested antibodies (CD99, FLI1, HNK1, and CAV1). CAV1 appeared as a diagnostic immunomarker of ES/PNET being positive in CD99-negative cases. Hence, the immunohistochemical analysis confirmed the diagnostic value of all four antibodies, which together cover more than 99% of the tumors, independently of the histological variety. The univariate analysis for survival revealed atypical ES as the only histological parameter apparently associated with less favorable clinical outcome, particularly in the subgroup of patients treated with surgery. In conclusion, the diagnosis of atypical ES is a challenge for the pathologist and needs support from molecular techniques to perform an optimal differential diagnosis with other small round cell tumors.


Occult HBV infection in HCC and cirrhotic tissue of HBsAg-negative patients: a virological and clinical study.
To evaluate the virological and clinical characteristics of occult HBV infection (OBI) in 68 consecutive HBsAg-negative patients with biopsy-proven cirrhosis and HCC. HBV DNA was sought and sequenced in plasma, HCC tissue and non-HCC liver tissue by PCRs using primers for HBV core, surface and x regions. OBI was identified by the presence of HBV DNA in at least two different PCRs. OBI was detected in HCC tissue of 13 (20%) patients and in non-HCC liver tissue of 3 of these 13. OBI was detected in HCC tissue of 54.5% of 11 anti-HBs- negative/anti-HBc-positive patients, in 29.4% of 17 anti-HBs/anti-HBc-positive and in 5% of 40 anti-HBs/anti-HBc-negative (p < 0.0005). The 13 patients with OBI in HCC tissue more frequently than the 55 without showed Child-B or -C cirrhosis (53.9% vs. 5.5%, p < 0.0001) and BCLC-B or -C stages (46.1% vs. 1.8%, p < 0.0001). The pre-S1, pre-S2 and S region sequences in HCC tissue showed amino acid (AA) substitutions (F19L, P24L, S59F, T131I, Q129H) and deletions (in positions 4,8, 17 and 86) in the S region, AA substitutions (T40S, P124K, L54P, G76A, N222T and I273L) in pre-S1 region and AA substitutions in pre-S2 region (P41H and P66L). In the 3 patients showing OBI also in non-HCC liver tissue the S, pre-S1 and pre-S2 sequencing displayed patterns of mutations different. The study showed a significant correlation between OBI and the severity of liver damage, several patterns of mutations in the S, pre-S1 and pre-S2 regions in HCC tissue, some at their first description.


Absolute and relative risk of cardiovascular disease in men with prostate cancer: results from the Population-Based PCBaSe Sweden.
Cardiovascular disease (CVD) is a potential adverse effect of endocrine treatment (ET) for prostate cancer (PC). We investigated absolute and relative CVD risk in 76,600 patients with PC undergoing ET, curative treatment, or surveillance. PCBaSe Sweden is based on the National Prostate Cancer Register, which covers more than 96% of PC cases. Standardized incidence ratios (SIRs) and standardized mortality ratios (SMRs) of ischemic heart disease (IHD), acute myocardial infarction (MI), arrhythmia, heart failure, and stroke were calculated to compare observed and expected (using total Swedish population) numbers of CVD, taking into account age, calendar time, and previous CVD. Between 1997 and 2007, 30,642 patients with PC received primary ET, 26,432 curative treatment, and 19,527 surveillance. SIRs for CVD were elevated in all men with the highest for those undergoing ET, independent of circulatory disease history (SIR MI for men without circulatory disease history: 1.40 [95% CI, 1.31 to 1.49], 1.15 [95% CI, 1.01 to 1.31], and 1.20 [95% CI, 1.11 to 1.30] for men undergoing ET, curative treatment, and surveillance, respectively). Absolute risk differences (ARD) showed that two (arrhythmia) to eight (IHD) extra cases of CVD would occur per 1,000 person-years. SMRs showed similar patterns, with ARD of zero (arrhythmia) to three (IHD) per 1,000 person-years. Increased relative risks of nonfatal and fatal CVD were found among all men with PC, especially those treated with ET. Because ET is currently the only effective treatment for metastatic disease and the ARDs were rather small, our findings indicate that CVD risk should be considered when prescribing ET but should not constitute a contraindication when the expected gain is tangible.


Association of cytomegalovirus genotype with graft rejection after liver transplantation.
The envelope glycoprotein gB of human cytomegalovirus (CMV) occurs as one of four main genotypes. Some previous studies have proposed a relationship of CMV gB genotype to the frequency of symptomatic infection and to clinical outcomes in both transplant and human immunodeficiency virus-infected populations. Our aim was to define the distribution of CMV gB genotypes and the impact on acute cellular rejection and graft/patient survival after orthotopic liver transplantation (OLT). Between October 1988 and December 1996, 325 patients underwent cyclosporine-based OLT at our center. CMV infection was surveyed prospectively and defined as viral isolation from blood or urine; 53 (16%) patients had detectable CMV. Isolates were genotyped by polymerase chain reaction amplification and restriction digest analysis. The distribution of CMV genotypes was: gB1, 19 (36%) patients; gB2, 15 (28%) patients; gB3, 13 (24%) patients; and gB4, 4 (8%) patients. Two patients (4%) had mixed infection (1 + 3, 1 + 4). Age, preOLT diagnosis, use of ganciclovir prophylaxis, basal immunosuppression, mean number of HLA donor/recipient mismatches, and United Network of Organ Sharing status were comparable among patients with different genotypes. Patients with gBl had a significantly higher mean number of acute rejection episodes (1.52+0.30 vs. 0.67+0.22; P=0.027). However, there was no difference in rejection severity, including OKT3 usage or FK506 conversion, or development of chronic rejection among patients with different genotypes. The gB genotype did not affect the development of symptomatic or tissue-invasive CMV disease, detected in 15 patients. Actuarial rates of patient (odds ratio [OR] 3.0; confidence interval [CI] 1.49-6.0) and graft (OR 2.57; CI 1.25-5.22) survival were significantly diminished in the group with CMV infection versus those without CMV (P<0.0001 for both), but there was no association with CMV genotype. (1) Patients with CMV infection had significantly reduced patient and graft survival rates at 1 and 5 years after OLT as compared with OLT recipients without CMV infection. (2) CMV genotype gB1 was associated with a higher mean number of acute rejection episodes.


Adenine nucleotide metabolism by chondrocytes in vitro: role of ATP in chondrocyte maturation and matrix mineralization.
The objective of the investigation was to explore the notion that chondrocytes in the growth plate secrete nucleotides and that these compounds are used to regulate cell maturation and matrix mineralization. Chondrocytes were isolated from the cephalic region of chick embryo sterna and maintained in culture until confluent. To promote expression of the mature phenotype, cultures were then treated with retinoic acid. During the culture period, medium was removed and analyzed for nucleotides using a modified reverse-phase high-performance liquid chromatography (HPLC) procedure. We found that culture medium, conditioned by the chondrocytes, contained significant quantities of nucleotides. Moreover, the nucleotide concentrations were similar in magnitude to levels reported for media conditioned by other cell types. In terms of species, adenosine diphosphate (ADP) was the major nucleotide present in the conditioned medium; adenosine monophosphate (AMP) was present, but at a lower concentration than ADP. To examine the possibility that adenosine triphosphate (ATP) was released by the cultured chondrocytes, but was rapidly degraded into ADP and AMP, we examined the kinetics of ATP breakdown by chondrocytes. We found that chondrocytes degraded over 70% of exogenous ATP within 15 minutes. Similar experiments performed with ADP and AMP indicated that these nucleotides were also degraded by the cells, but at a slower rate than ATP. To determine whether the extracellular nucleotides modulate cartilage development, we examined the effect of exogenous ATP on four major determinants of chondrocyte function: alkaline phosphatase activity, cell proliferation rate, anaerobic metabolism, and mineral deposition. We found that ATP caused only minimum alterations in cell number and alkaline phosphatase activity; however, it increased the lactate content of the medium probably by stimulating anaerobic glycolysis. We noted that ATP had a significant effect on the amount and type of mineral deposited into chondrocyte cultures. Compared with untreated controls, ATP stimulated formation of a small amount of poorly crystallized calcium phosphate. The results of the study show for the first time that chondrocytes release nucleotides into the extracellular milieu. Although they are rapidly degraded, they serve to regulate both mineral formation and energy metabolism.


Genetic syndromes and genetic tests in colorectal cancer.
Our understanding of the biology of colon cancer has matured to the point that it is a useful general paradigm for understanding solid tumor development. Recent advances provide insight into the genetic alterations underlying the development of colon cancer. These insights provide unique opportunities for genetic testing in predisposed, asymptomatic patients that can direct screening efforts and their clinical management. This review examines several inherited colon cancer predispositions, well described clinically for a century, that are now amenable to genetic testing. Additional discussion focuses on colon cancer predisposition traits that occur with high frequency but low penetrance characteristics. Finally, genetic tests for tumor markers that potentially have prognostic or therapeutic implications are reviewed.


Distinct somatic genetic changes associated with tumor progression in carriers of BRCA1 and BRCA2 germ-line mutations.
BRCA1 and BRCA2 mutations confer increased risk for development of breast cancer, but a number of additional, currently largely unknown, somatic genetic defects must also accumulate in the breast epithelial cells before malignancy develops. To evaluate the nature of these additional somatic genetic defects, we performed a genome-wide survey by comparative genomic hybridization on breast cancers from 21 BRCA1 mutation carriers, 15 BRCA2 mutation carriers, and 55 unselected controls. The total number of genetic changes was almost two times higher in tumors from both BRCA1 and BRCA2 mutation carriers than in the control group. In BRCA1 tumors, losses of 5q (86%), 4q (81%), 4p (64%), 2q (40%), and 12q (40%) were significantly more common than in the control group (7-13%). BRCA2 tumors were characterized by a higher frequency of 13q (73%) and 6q (60%) losses and gains of 17q22-q24 (87%) and 20q13 (60%) as compared to the prevalence of these changes in the control group (12-18%). In conclusion, accumulation of somatic genetic changes during tumor progression may follow a unique pathway in individuals genetically predisposed to cancer, especially by the BRCA1 gene. Activation or loss of genes in the affected chromosomal regions may be selected for during tumor progression in cells lacking functional BRCA1 or BRCA2. Identification of such genes could provide targets for therapeutic intervention and early diagnosis.