Inhibition activity of wild berry juice fractions against Streptococcus pneumoniae binding to human bronchial cells.
Bacterial adhesion to the cell surface is a crucial step before infection can take place. Inhibition of bacterial binding offers a novel preventive approach against infections. Cranberry (Vaccinium macrocarpon Ait.) juice has been found to have antiadhesive activity against different bacteria. Streptococcus pneumoniae is an important pathogen and the most common cause for pneumonia, meningitis, and otitis media. In this study the inhibitory activity of cranberry (Vaccinium oxycoccos L.), bilberry (Vaccinium myrtillus L.) and crowberry (Empetrum nigrum and Empetrum hermaphroditum L.) juice fractions against pneumococcal binding was tested using human bronchial cells (Calu-3) as an adhesion model. In addition, the antimicrobial activity of the berry juice fractions was tested. It was found that the studied berry juice fractions had antiadhesion activity and cranberry juice was the most active. The adhesion inhibition activity of cranberry juice was nearly 90% at a concentration of 8.7 mg/g of soluble solids. The antimicrobial activity of the studied berry juice fractions was found to be remarkable; pneumococcal growth was inhibited totally at a concentration of ∼86 mg/g. Both antiadhesion and antimicrobial activities were reduced after solid-phase extraction of the berry juices, which may suggest molecular synergistic effects of the berry juice molecules against S. pneumoniae. The findings indicate that cranberry, bilberry and crowberry juices have potential against pneumococcal infections.


Increased H-reflex excitability is not accompanied by changes in neural drive following 24 days of unilateral lower limb suspension.
The purpose of this study was to determine whether the gain in soleus H-reflex excitability induced by unilateral lower limb suspension (ULLS) is associated with changes in neural drive to the plantar flexor muscles. Six male subjects (23 ± 2 years, 187 ± 7 cm, 79 ± 9 kg) underwent 24 days of ULLS of the dominant limb. Plantar flexor maximal voluntary contraction (MVC) torque, activation capacity (twitch interpolation), soleus maximal electromyographic (EMG) activity, Hoffman (H)-reflex, and the first volitional (V) wave normalized to the compound muscle action potential (M-wave) were quantified before and after ULLS. Following ULLS, MVC torque decreased by 15% (P < 0.05). However, neither activation capacity nor EMG activity was significantly altered after the suspension. The V-wave remained unchanged consistently after ULLS, whereas the H-reflex increased significantly (+20%). Furthermore, there was no significant relationship between changes in H-reflex and V-wave over the ULLS period. These findings indicate that 24 days of ULLS can result in a substantial reduction of muscle strength without any apparent change in voluntary activation capacity. H-reflex and V-wave findings suggest that the spinal adaptations that underlie the unloading-induced increase in resting soleus H-reflex excitability did not significantly affect the efferent motor output to the plantar flexor muscles.


Trajectories of informal care and health.
The evidence of the impact of informal care provision on the health of carers presents a complex and contested picture, depending on the characteristics of the care studied, including its duration, which has been relatively short in previous research (up to 4 years). Drawing on data from the Office for National Statistics Longitudinal Study, a 1% sample of linked Census records for respondents in England and Wales (N=270,054), this paper contributes original insights on the impact of care provision on the carer's health ten years later. The paper explores differentials in self-reported health in 2011 between individuals according to their caring status at 2001 and 2011, and controlling for a range of demographic and socio-economic characteristics. The results show that individuals providing informal care in 2011 (regardless of carer status in 2001) exhibit lower odds of poor health in 2011 than those who did not provide care in both 2001 and 2011. Taking the intensity of care into account, 'heavy' carers in 2001 (i.e. caring for more than 20 h per week) who were not caring in 2011 show a higher likelihood of reporting poor health than non-carers, while those who were 'heavy' carers in both 2001 and 2011 are around one-third less likely to report poor health at 2011 compared to non-carers (2001 and 2011). These findings provide new insights in relation to repeat caring and its association with the carer's health status, further contributing to our understanding of the complex relationship between informal care provision and the carer's health.


Fabrication of nanoporous graphene/polymer composite membranes.
Graphene is currently investigated as a promising membrane material in which selective pores can be created depending on the requirements of the application. However, to handle large-area nanoporous graphene a stable support material is needed. Here, we report on composite membranes consisting of large-area single layer nanoporous graphene supported by a porous polymer. The fabrication is based on ion-track nanotechnology with swift heavy ions directly creating atomic pores in the graphene lattice and damaged tracks in the polymer support. Subsequent chemical etching converts the latent ion tracks in the supporting polymer foil, here polyethylene terephthalate (PET), into open microchannels while the perfectly aligned pores in the graphene top layer remain unaffected. To avoid unintentional damage creation and delamination of the graphene layer from the substrate, the graphene is encapsulated by a protecting poly(methyl methacrylate) (PMMA) layer. By this procedure a stable composite membrane is obtained consisting of nanoporous graphene (coverage close to 100%) suspended across selfaligned track-etched microchannels in a polymer support film. Our method presents a facile way to create high quality suspended graphene of tunable pore size supported on a flexible porous polymeric support, thus enabling the development of membranes for fast and selective ultrafiltration separation processes.


Sleep duration of underserved minority children in a cross-sectional study.
Short sleep duration has been shown to associate with increased risk of obesity. Childhood obesity is more prevalent among underserved minority children. The study measured the sleep duration of underserved minority children living in a large US urban environment using accelerometry and its relationship with BMI, socioeconomic status (SES), gender, ethnicity and physical activity. Time spent on sleep and physical activity among 333 Hispanic and 150 black children (9-12 y) was measured objectively by accelerometry over 5-7 consecutive days. The children were recruited at 14 underserved community centers in Houston, Texas, between January 2009 and February 2011. Body weight and height were measured in duplicate. The majority of children (88.8%) wore the monitor for 6 consecutive days. The children slept 8.8 ± 0.6 (mean ± SD) h/d and spent 45 ± 24 min/d on moderate-vigorous physical activity (MVPA). Hispanic children slept 0.2 h/d longer (P<0.001) than black children. Obese children slept 0.2 h/d less (P<0.02) than normal-weight children. SES had no effect on sleep duration. There was a significant interaction between gender and age (P<0.03); girls aged 11-12 y slept 0.3 h/d less than boys and the younger girls. Children slept 0.6 h/d longer (P<0.001) during the weekend than weekdays. No relation was detected between sleep duration and MVPA time. Minority children living in a large metropolitan area in the US are not meeting the National Sleep Foundation recommendation for sleep duration of 10-11 h/d. Longitudinal studies based on objective measures are needed to establish causality between sleep duration and obesity risk among minority children.


Genetic enhancement of fracture repair: healing of an experimental segmental defect by adenoviral transfer of the BMP-2 gene.
This study evaluated the ability of gene transfer to enhance bone healing. Segmental defects were created surgically in the femora of New Zealand white rabbits. First generation adenoviruses were used as vectors to introduce into the defects genes encoding either human bone morphogenetic protein-2 (BMP-2) or, as a negative control, firefly luciferase. Representative specimens were evaluated histologically after 8 weeks. Healing of the defects was monitored radiographically for 12 weeks, after which time the repair tissue was evaluated biomechanically. By radiological criteria, animals receiving the BMP-2 gene had healed their osseous lesions after 7 weeks, whereas those receiving the luciferase gene had not. Histologic examination of representative rabbits at 8 weeks confirmed ossification across the entire defect in response to the BMP-2 gene, whereas the control defect was predominantly fibrotic and sparsely ossified. At the end of the 12-week experiment, the control femora still showed no radiological signs of stable healing. The difference in radiologically defined healing between the experimental and control groups was statistically significant (P < 0. 002). Biomechanical testing of the femora at 12 weeks demonstrated statistically significant increases in the mean bending strength (P < 0.005) and bending stiffness (P < 0.05) of the animals treated with the BMP-2 gene. Direct, local adenoviral delivery of an osteogenic gene thus led to the healing of an osseous lesion that otherwise would not do so. These promising data encourage the further development of genetic approaches to enhancing bone healing. Gene Therapy (2000) 7, 734-739.


Quantitative Trait Loci Associated with Foliar Trigonelline Accumulation in Glycine Max L.
The objective of this study was to utilize a Glycine max RIL population to (1) evaluate foliar trigonelline (TRG) content in field-grown soybean, (2) determine the heritability of TRG accumulation, and (3) identify DNA markers linked to quantitative trait loci (QTLs) conditioning variation in TRG accumulation. Frequency distributions of 70 recombinant inbred lines showed statistically no significant departure from normality (P >.05) for TRG accumulation measured at pod development stage (R4). Six different molecular linkage groups (LGs) (B2, C2, D2, G, J, and K) were identified to be linked to QTLs for foliar TRG accumulation. Two unique microsatellite markers (SSR) on two different linkage groups identified QTL significantly associated with foliar TRG accumulation: a region on LG J (Satt285) (P =.0019, R(2) = 15.9%) and a second region on LG C2 (Satt079) (P =.0029, R(2) = 13.4%).


Dynamical systems approach to Saffman-Taylor fingering: dynamical solvability scenario.
A dynamical systems approach to competition of Saffman-Taylor fingers in a Hele-Shaw channel is developed. This is based on global analysis of the phase space flow of the low-dimensional ordinary-differential-equation sets associated with the classes of exact solutions of the problem without surface tension. Some simple examples are studied in detail. A general proof of the existence of finite-time singularities for broad classes of solutions is given. Solutions leading to finite-time interface pinchoff are also identified. The existence of a continuum of multifinger fixed points and its dynamical implications are discussed. We conclude that exact zero-surface tension solutions taken in a global sense as families of trajectories in phase space are unphysical because the multifinger fixed points are nonhyperbolic, and an unfolding does not exist within the same class of solutions. Hyperbolicity (saddle-point structure) of the multifinger fixed points is argued to be essential to the physically correct qualitative description of finger competition. The restoring of hyperbolicity by surface tension is proposed as the key point to formulate a generic dynamical solvability scenario for interfacial pattern selection.


Hatchet ribozyme structure and implications for cleavage mechanism.
Small self-cleaving ribozymes catalyze site-specific cleavage of their own phosphodiester backbone with implications for viral genome replication, pre-mRNA processing, and alternative splicing. We report on the 2.1-Å crystal structure of the hatchet ribozyme product, which adopts a compact pseudosymmetric dimeric scaffold, with each monomer stabilized by long-range interactions involving highly conserved nucleotides brought into close proximity of the scissile phosphate. Strikingly, the catalytic pocket contains a cavity capable of accommodating both the modeled scissile phosphate and its flanking 5' nucleoside. The resulting modeled precatalytic conformation incorporates a splayed-apart alignment at the scissile phosphate, thereby providing structure-based insights into the in-line cleavage mechanism. We identify a guanine lining the catalytic pocket positioned to contribute to cleavage chemistry. The functional relevance of structure-based insights into hatchet ribozyme catalysis is strongly supported by cleavage assays monitoring the impact of selected nucleobase and atom-specific mutations on ribozyme activity.


A Japanese-specific recurrent mutation and a novel splice site mutation in the LAMC2 gene identified in two Japanese families with Herlitz junctional epidermolysis bullosa.
Herlitz junctional epidermolysis bullosa (H-JEB) is an extremely rare genodermatosis characterized by lethality owing to severe blister formation. We report two unrelated Japanese patients with H-JEB. Genetic analyses detected a single nonsense mutation on the LAMC2 gene in these two patients. To identify the mutation involved and describe the first reported Japanese recurrent mutation in the LAMC2 gene. Direct sequencing was performed of DNA from either peripheral blood or fetal cells in amniotic fluid. Reverse transcriptase PCR was used to confirm that an aberrant transcript resulted from the splice site mutation. A haplotype analysis was performed to define the origin of the recurrent mutation. Both patients had blisters and erosions on the trunk and limbs at birth, with nail dystrophy. Patient 1 died as a result of sepsis at 30 weeks of age, and patient 2 died as a result of disseminated intravascular coagulation at 20 weeks of age. Mutation analysis of the LAMC2 gene revealed that patient 1 was compound heterozygous for a nonsense mutation (p.Cys553X) and a novel splice site mutation (c.2868+1delG), and patient 2 was a homozygous for p.Cys553X. Prenatal diagnosis performed during a subsequent pregnancy in family 2 revealed that this second child was heterozygous for p.Cys553X, and was thus not affected. Haplotype analysis suggested that a p.Cys553X allele derived from the same origin had been independently inherited by these two unrelated families. p.Cys553X in the LAMC2 gene may be a Japanese-specific recurrent mutation as a result of a founder effect, and it may therefore be useful for initial screening in the mutation analysis of H-JEB.


Agrofuels capitalism: a view from political economy.
This article considers the global expansion of agrofuels feedstock production from a political economy perspective. It considers and dismisses the environmental and pro-poor developmental justifications attached to agrofuels. To local populations and direct producers, the specific destination of the crop as fuel, food, cosmetics or other final uses in faraway places is probably of less interest than the forms of (direct or indirect) appropriation of their land and the forms of their insertion or exclusion as producers in global commodity chains. Global demand for both agrofuels and food is stimulating new forms (or the resurgence of old forms) of corporate land grabbing and expropriation, and of incorporation of smallholders in contracted production. Drawing both on recent studies on agrofuels expansion and on the political economy literature on agrarian transition and capitalism in agriculture, this article raises the question whether "agrofuels capitalism" is in any way essentially different from other forms of capitalist agrarian monocrop production, and in turn whether the agrarian transitions involved require new tools of analysis.


Prehospital electrocardiographic computer identification of ST-segment elevation myocardial infarction.
Identifying ST-segment elevation myocardial infarctions (STEMIs) in the field can decrease door-to-balloon times. Paramedics may use a computer algorithm to help them interpret prehospital electrocariograms (ECGs). It is unknown how accurately the computer can identify STEMIs. To Determine the sensitivity and specificity of prehospital ECGs in identifying patients with STEMI. Retrospective cross-sectional study of 200 prehospital ECGs acquired using Lifepak 12 monitors and transmitted by one of more than 20 emergency medical services (EMS) agencies to the emergency department (ED) of a Summa Akron City Hospital, a level 1 trauma center between January 1, 2007, and February 18, 2010. The ED sees more than 73,000 adult patients and treats 120 STEMIs annually. The laboratory performs 3,400 catheterizations annually. The first 100 patients with a diagnosis of STEMI and cardiac catheterization laboratory activation from the ED were analyzed. For comparison, a control group of 100 other ECGs from patients without a STEMI were randomly selected from our Medtronic database using a random-number generator. For patients with STEMI, an accurate computer interpretation was "acute MI suspected." Other interpretations were counted as misses. Specificity and sensitivity were calculated with confidence intervals (CIs). The sample size was determined a priori for a 95% CI of ±10%. Zero control patients were incorrectly labeled "acute MI suspected." The specificity was 100% (100/100; 95% CI 0.96-1.0), whereas the sensitivity was 58% (58/100; 95% CI 0.48-0.67). This would have resulted in 42 missed cardiac catheterization laboratory activations, but zero inappropriate activations. The most common incorrect interpretation of STEMI ECGs by the computer was "data quality prohibits interpretation," followed by "abnormal ECG unconfirmed." Prehospital computer interpretation is not sensitive for STEMI identification and should not be used as a single method for prehospital activation of the cardiac catheterizing laboratory. Because of its high specificity, it may serve as an adjunct to interpretation.


Energy Expenditure in Playground Games in Primary School Children Measured by Accelerometer and Heart Rate Monitors.
The aim of this study was to examine the energy expenditure (EE) measured using indirect calorimetry (IC) during playground games and to assess the validity of heart rate (HR) and accelerometry counts as indirect indicators of EE in children´s physical activity games. 32 primary school children (9.9 ± 0.6 years old, 19.8 ± 4.9 kg · m-2 BMI and 37.6 ± 7.2 ml · kg-1 · min-1 VO2max). Indirect calorimetry (IC), accelerometry and HR data were simultaneously collected for each child during a 90 min session of 30 playground games. Thirty-eight sessions were recorded in 32 different children. Each game was recorded at least in three occasions in other three children. The intersubject coefficient of variation within a game was 27% for IC, 37% for accelerometry and 13% for HR. The overall mean EE in the games was 4.2 ± 1.4 kcals · min-1 per game, totaling to 375 ± 122 kcals/per 90 min/session. The correlation coefficient between indirect calorimetry and accelerometer counts was 0.48 (p = .026) for endurance games and 0.21 (p = .574) for strength games. The correlation coefficient between indirect calorimetry and HR was 0.71 (p = .032) for endurance games and 0.48 (p = .026) for strength games. Our data indicate that both accelerometer and HR monitors are useful devices for estimating EE during endurance games, but only HR monitors estimates are accurate for endurance games.


Critically ill elderly who require mechanical ventilation: the effects of age on survival outcomes and resource utilisation in the medical intensive care unit of a general hospital.
Advanced age has been a criterion for denying admission to the medical intensive care unit (MICU) due to the perceived poorer outcome and increased resource utilisation. We studied the relationship between age and outcome of the critically ill mechanically-ventilated patients admitted to the MICU. This prospective study included patients admitted to the MICU for mechanical ventilation between 1994 and 1998. These were divided into 2 cohorts, with 206 patients aged 65 and above and 159 below 65 years. Outcome measures were MICU and hospital mortality and length of stay (LOS) in the MICU and hospital. Logistic and linear regression analyses were performed to determine the association between age and MICU and hospital mortality, as well as MICU and hospital LOS. Factors adjusted for included gender, smoking history, pre-hospitalisation functional status, ambulatory status, use of inotropes and APACHE II (m) scores (APACHE II scores were modified to exclude age points). Multivariate analysis revealed no statistically significant relationship between age and MICU or hospital mortality and LOS. However, APACHE IIM scores were significantly related to both MICU and hospital mortality (OR, 1.1; CI, 1.07-1.14 and OR, 1.1; CI, 1.09-1.18 respectively), but did not predict MICU or hospital LOS. Severity of acute illness and chronic co-morbidities, but not age, are predictors of MICU and hospital mortality in elderly ventilated patients.


The pain provocation-based straight leg raise test for diagnosis of lumbar disc herniation, lumbar radiculopathy, and/or sciatica: a systematic review of clinical utility.
The purpose of this study was to investigate the diagnostic accuracy of a finding of pain during the straight leg raise test for lumbar disc herniation, lumbar radiculopathy, and/or sciatica. The systematic review followed the PRISMA guidelines and captured articles that had variable designs, standardized reference standards, described an index test of a pain response, and that reported both sensitivity and specificity for the straight leg raise test. Each article specified that a positive straight leg raise test was one in which the subject felt pain during the administration of the test, and was assessed for quality using the QUADAS tool. Sensitivities and specificities were evaluated along with positive and negative likelihood ratios. The systematic review and hand search identified 7 articles for inclusion; three of these articles were rated as high quality using QUADAS scores. Sensitivity and specificity varied among the 7 studies with 4 suggesting that a pain response SLR is sensitive whereas 3 suggested it is a specific measure. Variability in reference standard may partly explain the inconsistencies in the diagnostic accuracy findings. Further, pain that is not specific to lumbar radiculopathy, such as that associated with hamstring tightness, may also lead to false positives for the SLR; and may inflate the sensitivity of the test.


Intermittent control strategy can enhance stabilization robustness in bumblebee hovering.
Active flight control plays a crucial role in stabilizing the body posture of insects to stay aloft under a complex natural environment. Insects can achieve a closed-loop flight control by integrating the external mechanical system and the internal working system through manipulating wing kinematics according to feedback information from multiple sensors. While studies of proportional derivative/proportional integral derivative-based algorithms are the main subject to explore the continuous flight control mechanisms associated with insect flights, it is normally observed that insects achieve an intermittent spike firing in steering muscles to manipulate wings in flight control discontinuously. Here we proposed a novel intermittent control strategy for a 3 degree of freedom (DoF) pitch-control and explored its stabilization robustness in bumblebee hovering. An integrated computational model was established and validated, which comprises an insect-inspired dynamic flight simulator and a novel discrete feedback controller as well as a simplified free-flight dynamic model. We found that the intermittent control model can achieve an angular-dominant flight control, whereas the continuous control model corresponds to an angular-velocity-dominant one. Given the biological constraints in sensorimotor neurobiology and musculoskeletal mechanics, the intermittent control strategy was examined capable of enhancing the stabilization robustness in terms of sensory latency, stroke derivation, spike interval, and damping strength. Our results indicate that the intermittent control strategy is likely a sophisticated flight control mechanism in insect flights while providing a bioinspired flight-control design for insect size flapping-wing micro air vehicles.


Absence of antibody to HTLV I and III in sera of Canadian patients with multiple sclerosis and chronic myelopathy.
Recently, elevated titers of antibody to HTLV I have been demonstrated in patients with tropical spastic paraparesis and multiple sclerosis. We evaluated the possible role of human retroviruses HTLV I and III in Canadian patients with multiple sclerosis and chronic idiopathic myelopathy. Using sensitive enzyme immunoassays, we were unable to find antibody to either HTLV I or III in 201 patients with multiple sclerosis, 29 patients with chronic myelopathy, 51 patients with other neurological disorders, or 29 normal subjects. These data do not support a role for these viruses in the cause of sporadic multiple sclerosis and chronic myelopathy in a regionally based Canadian population, but do not exclude a role for other antigenically distinct retroviruses.


Alkaloid chemophenetics and transcriptomics of the Nicotiana genus.
In this study, we determined the pyridine alkaloid content (nicotine, nornicotine, anabasine, anatabine, cotinine, and myosmine) of 58 species and 2 subspecies of the Nicotiana genus by ultra-high-performance liquid chromatography coupled with mass spectrometry. We observed clear correlation between Noctiflorae and Suaveolentes sections and their above average accumulation of anabasine in the genus. In addition, the results demonstrated the presence of not only trace amounts but quantifiable levels of myosmine, an alkaloid previously detected in only minute quantities, in the leaves and roots of 16 species. In this study, analysis of gene expression of 58 species and 2 subspecies from the Nicotiana genus by mRNA sequencing was performed for the first time. Sequencing reads were mapped against annotated genes of a Nicotiana tabacum reference genome and expression values were subsequently calculated. Hierarchical clustering of alkaloid biosynthesis pathway genes and alkaloid content composition revealed patterns clearly segregating Nicotiana sections. Correlation of gene expression with alkaloid accumulation phenotypes was evident, including low putrescine methyltransferase expression for all species in the Suaveolentes section or clear correlation of nicotine demethylase with conversion rates of nicotine to nornicotine in the majority of species. Multiple additional correlations between alkaloid accumulation and gene expression values were identified, which makes this study an important fundament toward future scientific exploration of the Nicotiana genus.


Studies on the structure of muscle. III. Phase contrast and electron microscopy of dipteran flight muscle.
1. The flight muscles of blowflies are easily dispersed in appropriate media to form suspensions of myofibrils which are highly suitable for phase contrast observation of the band changes associated with ATP-induced contraction. 2. Fresh myofibrils show a simple band pattern in which the A substance is uniformly distributed throughout the sarcomere, while the pattern characteristic of glycerinated material is identical with that generally regarded as typical of relaxed vertebrate myofibrils (A, I, H, Z, and M bands present). 3. Unrestrained myofibrils of both fresh and glycerinated muscle shorten by not more than about 20 per cent on exposure to ATP. In both cases the A substance migrates during contraction and accumulates in dense bands in the Z region, while material also accumulates in the M region. It is proposed that these dense contraction bands be designated the C(z), and C(m) bands respectively. In restrained myofibrils, the I band does not disappear, but the C(z) and C(m) bands still appear in the presence of ATP. 4. The birefringence of the myofibrils decreases somewhat during contraction, but the shift of A substance does not result in an increase of birefringence in the C(z) and C(m) bands. It seems therefore that the A substance, if it is oriented parallel with the fibre axis in the relaxed myofibril, must exist in a coiled or folded configuration in the C hands of contracted myofibrils. 5. The fine structure of the flight muscle has been determined from electron microscopic examination of ultrathin sections. The myofibrils are of roughly hexagonal cross-section and consist of a regular single hexagonal array of compound myofilaments the cores of which extend continuously throughout all bands of the sarcomere in all states of contraction or relaxation so far investigated. 6. Each myofilament is joined laterally with its six nearest neighbours by thin filamentous bridges which repeat at regular intervals along the fibre axis and are present in the A, I, and Z, but not in the H or M bands. When stained with PTA, the myofilaments display a compound structure. In the A band, a lightly staining medullary region about 40 A in diameter is surrounded by a densely staining cortex, the over-all diameter of the myofilament being about 120 A. This thick cortex is absent in the I and H bands, but a thinner cortex is often visible. 7. It is suggested that the basic structure is a longitudinally continuous framework of F actin filaments, which are linked periodically by the lateral bridges (possibly tropomyosin). The A substance is free under certain conditions to migrate to the Z bands to form the C(z) bands. The material forming the C(m) bands possibly represents another component of the A substance. The results do not clearly indicate whether myosin is confined to the A bands or distributed throughout the sarcomere.


A novel mouse model to study the virulence of and host response to Porphyromonas (Bacteroides) gingivalis.
We describe here the development of a mouse subcutaneous chamber model that allows for the examination of host-parasite interactions as well as the determination of gross pathology with Porphyromonas (Bacteroides) gingivalis challenge. When inoculated into stainless-steel chambers implanted subcutaneously in female BALB/c mice, P. gingivalis W83, W50, and A7436 (10(8) to 10(10) CFU) caused cachexia, ruffling, general erythema and phlegmonous, ulcerated, necrotic lesions, and death. P. gingivalis W50/BEI, HG405, and 33277 (10(10) CFU) produced localized abscesses in the mouse chamber model with rejection of chambers at the injection site. Analysis of chamber fluid from 33277-, HG405-, and W50/BEI-infected mice by cytocentrifugation revealed inflammatory cell debris, polymorphonuclear leukocytes, and high numbers of dead bacteria. In contrast, fluid from A7436-, W50-, and W83-infected mice revealed infiltration predominantly of polymorphonuclear leukocytes and live bacteria. Bacteria were found primarily associated with polymorphonuclear leukocytes in the fluid from W50-, HG405-, and W83-infected mice but not from A7436-infected mice. Viable isolates were recoverable from the chamber fluid through day 3 for W50/BEI, day 5 for 33277, day 6 for HG405, day 7 for W50, day 14 for W83, and day 26 for A7436. All strains induced a systemic immunoglobulin G response in serum and chamber fluid samples. The use of this model will allow us to examine the virulence of P. gingivalis as defined by the interaction of host response to localized infection with P. gingivalis.


Sampling ARG of multiple populations under complex configurations of subdivision and admixture.
Simulating complex evolution scenarios of multiple populations is an important task for answering many basic questions relating to population genomics. Apart from the population samples, the underlying Ancestral Recombinations Graph (ARG) is an additional important means in hypothesis checking and reconstruction studies. Furthermore, complex simulations require a plethora of interdependent parameters making even the scenario-specification highly non-trivial. We present an algorithm SimRA that simulates generic multiple population evolution model with admixture. It is based on random graphs that improve dramatically in time and space requirements of the classical algorithm of single populations.Using the underlying random graphs model, we also derive closed forms of expected values of the ARG characteristics i.e., height of the graph, number of recombinations, number of mutations and population diversity in terms of its defining parameters. This is crucial in aiding the user to specify meaningful parameters for the complex scenario simulations, not through trial-and-error based on raw compute power but intelligent parameter estimation. To the best of our knowledge this is the first time closed form expressions have been computed for the ARG properties. We show that the expected values closely match the empirical values through simulations.Finally, we demonstrate that SimRA produces the ARG in compact forms without compromising any accuracy. We demonstrate the compactness and accuracy through extensive experiments. SimRA (Simulation based on Random graph Algorithms) source, executable, user manual and sample input-output sets are available for downloading at: https://github.com/ComputationalGenomics/SimRA CONTACT: : firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


The doublecortin-expressing population in the developing and adult brain contains multipotential precursors in addition to neuronal-lineage cells.
Doublecortin (DCX) has recently been promulgated as a selective marker of cells committed to the neuronal lineage in both the developing and the adult brain. To explore the potential of DCX-positive (DCX+) cells more stringently, these cells were isolated by flow cytometry from the brains of transgenic mice expressing green fluorescent protein under the control of the DCX promoter in embryonic, early postnatal, and adult animals. It was found that virtually all of the cells (99.9%) expressing high levels of DCX (DCX(high)) in the embryonic brain coexpressed the neuronal marker betaIII-tubulin and that this population contained no stem-like cells as demonstrated by lack of neurosphere formation in vitro. However, the DCX+ population from the early postnatal brain and the adult subventricular zone and hippocampus, which expressed low levels of DCX (DCX(low)), was enriched for neurosphere-forming cells, with only a small subpopulation of these cells coexpressing the neuronal markers betaIII-tubulin or microtubule-associated protein 2. Similarly, the DCX(low) population from embryonic day 14 (E14) brain contained neurosphere-forming cells. Only the postnatal cerebellum and adult olfactory bulb contained some DCX(high) cells, which were shown to be similar to the E14 DCX(high) cells in that they had no stem cell activity. Electrophysiological studies confirmed the heterogeneous nature of DCX+ cells, with some cells displaying characteristics of immature or mature neurons, whereas others showed no neuronal characteristics whatsoever. These results indicate that DCX(high) cells, regardless of location, are restricted to the neuronal lineage or are bone fide neurons, whereas some DCX(low) cells retain their multipotentiality.


Long-term Proton Pump Inhibitor Administration Caused Physiological and Microbiota Changes in Rats.
Proton pump inhibitors (PPIs) are used for the long-term treatment of gastroesophageal disorders and the non-prescription medicines for acid reflux. However, there is growing concerns about PPI misuse, overuse and abuse. This study aimed to develop an animal model to examine the effects of long-term use of PPI in vivo. Twenty one Wistar rats were given omeprazole orally or intravenously for 30 days, and caerulein as a positive control. After euthanization, the serum and stool were collected to perform MS-based quantitative analysis of metabolites. We carried out 16S-based profiling of fecal microbiota, assessed the expression of bile acid metabolism regulators and examined the immunopathological characteristics of bile ducts. After long-term PPI exposure, the fecal microbial profile was altered and showed similarity to those observed in high-fat diet studies. The concentrations of several metabolites were also changed in various specimens. Surprisingly, morphological changes were observed in the bile duct, including ductal epithelial proliferation, micropapillary growth of biliary epithelium, focal bile duct stricture formation and bile duct obstruction. These are characteristics of precancerous lesions of bile duct. FXR and RXRα expressions were significantly reduced, which were similar to that observed in cholangiocarcinoma in TCGA and Oncomine databases. We established a novel animal model to examine the effects of long-term use of omeprazole. The gut microbes and metabolic change are consequences of long-term PPI exposure. And the results showed the environment in vivo tends to a high-fat diet. More importantly, we observed biliary epithelial hyperplasia, which is an indicator of a high-fat diet.


Congenital malaria and neonatal bacterial co-infection in twins prematurely born to a mother with sickle-cell anaemia in the Democratic Republic of the Congo.
We report cases of gestational and congenital malaria with twin prematurity, low birth weight and bacterial co-infection. Congenital malaria is often misdiagnosed for lack of specific symptoms and a general lack of awareness of this presumably uncommon condition, and its diagnosis and prognosis become even more complex in the event of bacterial co-infections. A 35-weeks pregnant woman with sickle-cell disease and a history of spontaneous abortions was admitted at Vanga Hospital in DR Congo. She had fever (38.9°C) and microscopy-confirmed P. falciparum malaria and was put on 80/480 mg artemether-lumefantrine. She soon went into active labour, during which both twins developed acute foetal distress and were promptly delivered by C-section. The twins were underweight, and both had P. falciparum malaria at birth and were given 20 mg quinine twice daily. Both developed fever on the third day; a bacterial infection was suspected and 200 mg ceftriaxone was added to their treatment. Fever in both twins quickly resolved, and one twin totally recovered within 2 days of antibiotic treatment. The other twin developed acute respiratory distress and hypoxia and died. This is a case of gestational and congenital malaria with prematurity, low birth weight and bacterial co-infection, but the patients were initially only treated for malaria based on their malaria-positive blood smears at birth. In malaria-endemic areas, babies should be screened for congenital malaria. Even with a confirmed malaria infection in the new-born, it is important consider the possibility of bacterial co-infections.


Proteomic analysis of post-hemorrhagic shock mesenteric lymph.
Recent studies have documented the association of mesenteric lymphatic route with adult respiratory distress syndrome and multiple organ failure after hemorrhagic shock. However, the mediators and mechanisms of the toxic effects of mesenteric lymph remain unclear. This study aimed to identify mediators or biomarkers in the mesenteric lymph through comparative proteomic analysis. Fourteen mature male Sprague-Dawley rats were randomly divided and subjected to trauma (laparotomy) plus hemorrhagic shock or trauma plus sham shock. Mesenteric lymph samples were collected before shock and at 3 h after resuscitation from hemorrhagic shock (or sham shock). To investigate changes in proteome profiles between preshock and 3-h postshock (or 3-h post-sham shock) mesenteric lymph samples, two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were performed. We found a more than 2-fold change in abundance of 31 protein spots in the lymph samples. Mass spectrometry analyses identified 12 distinct proteins. Four proteins were consistently upregulated in the 3-h postshock lymph samples, including serum albumin precursor, two isoforms of cytoplasmic actin, complement C3 precursor, and major urinary protein precursor. Two proteins, including haptoglobin and one unidentified protein, were consistently downregulated. The deregulation of these proteins was confirmed by Western blots. Most of these altered proteins are functionally implicated in tissue inflammation. The findings of this study provide a starting point for investigating the functions of these proteins in hemorrhagic shock-induced lung injury and hold great promise for the development of potential therapeutic interventions.


Intravenous supplementation type and volume are associated with 1-year outcome and major complications in patients with chronic intestinal failure.
No marker to categorise the severity of chronic intestinal failure (CIF) has been developed. A 1-year international survey was carried out to investigate whether the European Society for Clinical Nutrition and Metabolism clinical classification of CIF, based on the type and volume of the intravenous supplementation (IVS), could be an indicator of CIF severity. At baseline, participating home parenteral nutrition (HPN) centres enrolled all adults with ongoing CIF due to non-malignant disease; demographic data, body mass index, CIF mechanism, underlying disease, HPN duration and IVS category were recorded for each patient. The type of IVS was classified as fluid and electrolyte alone (FE) or parenteral nutrition admixture (PN). The mean daily IVS volume, calculated on a weekly basis, was categorised as <1, 1-2, 2-3 and >3 L/day. The severity of CIF was determined by patient outcome (still on HPN, weaned from HPN, deceased) and the occurrence of major HPN/CIF-related complications: intestinal failure-associated liver disease (IFALD), catheter-related venous thrombosis and catheter-related bloodstream infection (CRBSI). Fifty-one HPN centres included 2194 patients. The analysis showed that both IVS type and volume were independently associated with the odds of weaning from HPN (significantly higher for PN <1 L/day than for FE and all PN >1 L/day), patients' death (lower for FE, p=0.079), presence of IFALD cholestasis/liver failure and occurrence of CRBSI (significantly higher for PN 2-3 and PN >3 L/day). The type and volume of IVS required by patients with CIF could be indicators to categorise the severity of CIF in both clinical practice and research protocols.


Modulation response of a long-cavity, gain-levered quantum-dot semiconductor laser.
The gain-lever effect enhances the modulation efficiency of a semiconductor laser when compared to modulating the entire laser. This technique is investigated in a long-cavity multi-section quantum-dot laser where the length of the modulation section is varied to achieve 14:2, 15:1 and 0:16 gain-to-modulation section ratios. In this work, the gain-levered modulation configuration resulted in an increase in modulation efficiency by as much as 16 dB. This investigation also found that the 3-dB modulation bandwidth and modulation efficiency are dependent on the modulation section length of the device, indicating the existence of an optimal gain-to-modulation section ratio. The long cavity length of the multi-section laser yielded a distinctive case where characteristics of both the gain-lever effect and spatial effects are observed in the modulation response. Here, spatial effects within the cavity dominated the small-signal modulation response close to and above the cavity's free-spectral range frequency, whereas the gain-lever effect influenced the modulation response throughout the entirety of the response.


Telomere length in early childhood: Early life risk factors and association with carotid intima-media thickness in later childhood.
Reduced telomere length is a measure of biological aging that is predictive of cardiac events in adults, and has been mechanistically implicated in the onset and progression of atherosclerosis. We sought to describe the early life factors associated with leukocyte telomere length in early childhood, and to determine whether telomere length measured during early childhood is associated with arterial wall thickening later in childhood. A longitudinal birth cohort recruited antenatally in Sydney from 1997 to 1999. Leukocyte telomere length was measured in 331 children at age 3.6 years (SD 1.0); of whom 268 children without diabetes had carotid intima-media thickness assessed by ultrasound at age 8 years. Male sex, younger paternal age and higher maternal body mass index were associated with shorter telomere length in early childhood, which in turn was associated with greater carotid intima-media thickness at age 8 years (standardised β = -0.159, P = 0.01). There was a graded association across quartiles of telomere length (Ptrend = 0.001) with the highest odds of elevated intima-media thickness (>75th percentile) being in children with the shortest telomeres (odds ratio 4.00 (95% confidence interval 1.58 to 10.14) relative to those with the longest telomeres, P = 0.003). This association remained after adjustment for early life risk factors (Ptrend = 0.001). Reduced telomere length in early childhood is independently associated with arterial wall thickness in later childhood, suggesting that reduced telomere length during early childhood may be a marker of vascular disease risk.


Patterns of childhood social development in adult schizophrenics.
Previous analysis of teachers' comments in school records of schizophrenics showed progressive deviance in their childhood social behavior and sharp differences between the sexes. This report presents a replication of the longitudinal study and a reanalysis of the combined samples. The principal findings from the earlier study were confirmed: in the later school years, preschizophrenic girls were primarily introverted and preschizophrenic boys mainly disagreeable. In the new study, the preschizophrenic girls were also emotionally unstable, introverted, and passive from kindergarten through grade 6, suggesting in earlier history of social disengagement than was previously apparent, and the preschizophrenic boys were emotionally unstable in grades 7 through 12, confirming a nonsignificant trend in the original study. The boys were not behaviorally distinguishable from other boys in elementary grades, so the notion of progressive deviance appears more applicable in their case.


Health Disparities and Converging Epidemics in Jail Populations: Protocol for a Mixed-Methods Study.
Incarcerated populations have increased in the last 20 years and >12 million individuals cycle in and out of jails each year. Previous research has predominately focused on the prison population. However, a substantial gap exists in understanding the health, well-being, and health care utilization patterns in jail populations. This pilot study has 5 main objectives: (1) define recidivists of the jail system, individuals characterized by high incarceration rates; (2) describe and compare the demographic and clinical characteristics of incarcerated individuals; (3) identify jail-associated health disparities; (4) estimate associations between incarceration and health; and (5) describe model patterns in health care and jail utilization. The project has two processes-a secondary data analysis and primary data collection-which includes a cross-sectional health survey and biological sample collection to investigate infectious disease characteristics of the jail population. This protocol contains pilot elements in four areas: (1) instrument validity and reliability; (2) individual item assessment; (3) proof of concept of content and database accessibility; and (4) pilot test of the "honest broker" system. Secondary data analysis includes the analysis of 6 distinct databases, each covered by a formal memorandum of agreement between Northern Arizona University and the designated institution: (1) the Superior Court of Arizona Public Case Finder database; (2) North Country Health Care; (3) Health Choice Integrated Care; (4) Criminal Justice Information Services; (5) Correctional Electronic Medical Records; and (6) iLEADS. We will perform data integration processes using an automated honest broker design. We will administer a cross-sectional health survey, which includes questions about health status, health history, health care utilization, substance use practices, physical activity, adverse childhood events, and behavioral health, among 200 Coconino County Detention Facility inmates. Concurrent with the survey administration, we will collect Methicillin-resistant and Methicillin-sensitive Staphylococcus aureus (samples from the nose) and dental microbiome (Streptococcus sobrinus and Streptococcus mutans samples from the mouth) from consenting participants. To date, we have permission to link data across acquired databases. We have initiated data transfer, protection, and initial assessment of the 6 secondary databases. Of 199 inmates consented and enrolled, we have permission from 97.0% (193/199) to access and link electronic medical and incarceration records to their survey responses, and 95.0% (189/199) of interviewed inmates have given nasal and buccal swabs for analysis of S. aureus and the dental microbiome. This study is designed to increase the understanding of health needs and health care utilization patterns among jail populations, with a special emphasis on frequently incarcerated individuals. Our findings will help identify intervention points throughout the criminal justice and health care systems to improve health and reduce health disparities among jail inmates. RR1-10.2196/10337.


Building consensus on definition and nomenclature of hepatic, pancreatic, and biliary organoids.
Hepatic, pancreatic, and biliary (HPB) organoids are powerful tools for studying development, disease, and regeneration. As organoid research expands, the need for clear definitions and nomenclature describing these systems also grows. To facilitate scientific communication and consistent interpretation, we revisit the concept of an organoid and introduce an intuitive classification system and nomenclature for describing these 3D structures through the consensus of experts in the field. To promote the standardization and validation of HPB organoids, we propose guidelines for establishing, characterizing, and benchmarking future systems. Finally, we address some of the major challenges to the clinical application of organoids.


Characterization of the Lytic Bacteriophage phiEaP-8 Effective against Both Erwinia amylovora and Erwinia pyrifoliae Causing Severe Diseases in Apple and Pear.
Bacteriophages, bacteria-infecting viruses, have been recently reconsidered as a biological control tool for preventing bacterial pathogens. Erwinia amylovora and E. pyrifoliae cause fire blight and black shoot blight disease in apple and pear, respectively. In this study, the bacteriophage phiEaP-8 was isolated from apple orchard soil and could efficiently and specifically kill both E. amylovora and E. pyrifoliae. This bacteriophage belongs to the Podoviridae family. Whole genome analysis revealed that phiEaP-8 carries a 75,929 bp genomic DNA with 78 coding sequences and 5 tRNA genes. Genome comparison showed that phiEaP-8 has only 85% identity to known bacteriophages at the DNA level. PhiEaP-8 retained lytic activity up to 50°C, within a pH range from 5 to 10, and under 365 nm UV light. Based on these characteristics, the bacteriophage phiEaP-8 is novel and carries potential to control both E. amylovora and E. pyrifoliae in apple and pear.


Exercise and neuroplasticity in persons living with Parkinson's disease.
For many years, exercise was not a recommended rehabilitation strategy for persons with a diagnosis of idiopathic Parkinson's disease (PD). Since it was believed that exercise had no measurable effect on PD, or might worsen the underlying pathology, it was to be avoided. A rich vein of bench and translational research now suggest non-pharmacological approaches, such as exercise or physiotherapy, have a far greater effect on the cardinal features of PD than previously believed. In particular, recent studies utilizing animal models of PD have begun to explore the molecular mechanisms of exercise-induced changes in the pathophysiology of PD. Yet, many clinicians and communities remain unaware of the scientific literature underlying exercise-induced brain repair or reorganization (neuroplasticity) and accompanying behavioral recovery in animal models of PD. The authors will summarize some noteworthy preliminary studies suggesting that continuous, deficit targeted, intensive training may confer neuroprotection and thereby, slow, stop or reverse the progression of the disease or promote neurorestoration through adaptation of compromised signaling pathways. While much work remains and these preliminary results await replication in larger prospective human trials, we believe a major challenge in the field of non-pharmacological, rehabilitative intervention for PD will be the extent to which healthcare providers are able to translate the science of exercise and PD to the level of the community.


Effect of slightly lowered body temperatures on endurance performance in humans.
Eight well-trained male rowers exercised as hard as possible on a bicycle ergometer for 60 min at an ambient temperature of 18 degrees C. On 1 day (precooling test, PRET) exercise was preceded by a double cold exposure that caused starting body temperatures to be approximately 4.5, 1.0, 0.8, and 0.4 degrees C (mean skin, mean body, tympanic, and esophageal temperatures, respectively) lower than in the control test (CONT). In PRET the mean 1-h work rate (172 W) was 6.8% larger than in CONT (161 W), O2 uptake (Vo2) was 9.6% higher (2.86 vs. 2.61 1 X min-1), and O2 pulse was increased by 5.6% (18.8 vs. 17.8 ml), whereas the sweat rate was 20.3% lower in PRET (1.06 vs. 1.33 mg X cm-2.min-1). No differences in heart rate, efficiency, postexercise blood values of lactate, or acid-base status were demonstrated. It appears that the improved performance in PRET was related to an increased O2 supply to the working muscles. Although plasma levels of total beta H-endorphin immunoreactivity were in the same range under both test conditions, different components of beta H-endorphin immunoreactivity were indicated to exist in PRET and CONT.


The Multiple Sclerosis Impact Scale (MSIS-29): a new patient-based outcome measure.
Changes in health policy have underlined the importance of evidence-based clinical practice and rigorous evaluation of patient-based outcomes. As patient-based outcome measurement is particularly important in treatment trials of multiple sclerosis, a number of disease-specific instruments have been developed recently. One limitation of these instruments is that none was developed using the standard psychometric approach of reducing a large item pool generated from people with multiple sclerosis. Consequently, an outcome measure for clinical trials of multiple sclerosis that is disease specific and combines patient perspective with rigorous psychometric methods will complement existing instruments. The aim of this study was to develop such a measure. Standard psychometric methods were used. A pool of 129 questionnaire items was generated from interviews with 30 people with multiple sclerosis, expert opinion and literature review. The questionnaire was administered by postal survey to 1530 people selected randomly from the Multiple Sclerosis Society membership database. Redundant items and those with limited measurement properties were removed. The remaining items (n = 41) were grouped into scales using factor analysis, and then refined to form the Multiple Sclerosis Impact Scale (MSIS-29), an instrument measuring the physical (20 items) and psychological (nine items) impact of multiple sclerosis. Five psychometric properties of the MSIS-29 (data quality, scaling assumptions, acceptability, reliability and validity) were examined in a separate postal survey of 1250 Multiple Sclerosis Society members. A preliminary responsiveness study of the MSIS-29 was undertaken in 55 people admitted for rehabilitation and intravenous steroid treatment of relapses. The MSIS-29 satisfied all psychometric criteria. Data quality was excellent, missing data were low (maximum 3.9%), item test-re-test reliability was high (r = 0.65-0.90) and scale scores could be generated for >98% of respondents. Item descriptive statistics, item convergent and discriminant validity, and factor analysis indicated that it was legitimate to generate scores for MSIS-29 scales by summing items. MSIS-29 scales showed good variability, small floor and ceiling effects, high internal consistency (Cronbach's alpha <or=0.91) and high test-re-test reliability (intraclass correlation <or=0.87). Correlations with other measures and the analysis of group differences provided evidence that the MSIS-29 measures the physical and psychological impact of multiple sclerosis. Effect sizes (physical scale = 0.82, psychological scale = 0.66) demonstrated preliminary evidence of good responsiveness. These results indicate the MSIS-29 is a clinically useful and scientifically sound patient-based outcome measure of the impact of multiple sclerosis suitable for clinical trials and epidemiological studies.


A Web-Based Therapeutic Program (We Can Do This) for Reducing Methamphetamine Use and Increasing Help-Seeking Among Aboriginal and Torres Strait Islander People: Protocol for a Randomized Wait-List Controlled Trial.
Methamphetamine use is of deep concern to Aboriginal and Torres Strait Islander communities, but access to culturally appropriate treatment resources and services is limited. Web-based programs have potential as flexible and cost-effective additions to the range of treatment options available to Aboriginal people. The We Can Do This online intervention is designed to incorporate evidence-based therapies in a culturally relevant format using narratives from Aboriginal people to contextualize the therapeutic content. The goal of the research will be to test the effectiveness of the online intervention in a wait-list controlled randomized trial across multiple sites in urban, regional, and remote locations. Participants will be Aboriginal and Torres Strait Islander people aged 16 years and over who have used methamphetamine at least weekly for the previous 3 months. They will be recruited online and via health services. During the intervention phase, participants will have access to the online intervention for 6 weeks with optional telephone or face-to-face support provided by participating health services. The primary outcome measure will be the number of days the participant used methamphetamine over the past 4 weeks compared to wait-list controls, assessed at baseline, 1, 2, and 3 months. Secondary outcomes will include help-seeking, readiness to change, severity of dependence, and psychological distress. Any important changes to the protocol will be agreed upon by the trial management committee and communicated to all relevant parties, including trial site representatives and the trial registry. Recruitment will commence in July 2019, and results are expected in early 2021. This research is funded by National Health and Medical Research Council project grant #1100696. The primary sponsor for the trial is the South Australian Health and Medical Research Institute. A trial management committee with representation from the participating health services, chief investigators, other Aboriginal experts, and consumers will oversee procedures, trial conduct, analysis, and reporting of the results. The trial of this online intervention builds on existing research supporting the effectiveness of Web-based therapies for a range of psychological and other health-related issues including substance use. If successful, the We Can Do this online intervention will increase the range of options available to Aboriginal people seeking to reduce or stop methamphetamine use. It may provide a pathway into treatment for people who may otherwise be disengaged with health services for a range of reasons and will be a culturally appropriate, evidence-based resource for health practitioners to offer their clients. Australian New Zealand Clinical Trials Registry ACTRN12619000134123p; https://www.anzctr.org.au/ Trial/Registration/TrialReview.aspx?id=376088&isReview=true. PRR1-10.2196/14084.


Variability in lutetium-177 SPECT quantification between different state-of-the-art SPECT/CT systems.
Quantitative SPECT imaging in targeted radionuclide therapy with lutetium-177 holds great potential for individualized treatment based on dose assessment. The establishment of dose-effect relations requires a standardized method for SPECT quantification. The purpose of this multi-center study is to evaluate quantitative accuracy and inter-system variations of different SPECT/CT systems with corresponding commercially available quantitative reconstruction algorithms. This is an important step towards a vendor-independent standard for quantitative lutetium-177 SPECT. Four state-of-the-art SPECT/CT systems were included: Discovery™ NM/CT 670Pro (GE Healthcare), Symbia Intevo™, and two Symbia™ T16 (Siemens Healthineers). Quantitative accuracy and inter-system variations were evaluated by repeatedly scanning a cylindrical phantom with 6 spherical inserts (0.5 - 113 ml). A sphere-to-background activity concentration ratio of 10:1 was used. Acquisition settings were standardized: medium energy collimator, body contour trajectory, photon energy window of 208 keV (± 10%), adjacent 20% lower scatter window, 2 × 64 projections, 128 × 128 matrix size, and 40 s projection time. Reconstructions were performed using GE Evolution with Q.Metrix™, Siemens xSPECT Quant™, Siemens Broad Quantification™ or Siemens Flash3D™ algorithms using vendor recommended settings. In addition, projection data were reconstructed using Hermes SUV SPECT™ with standardized reconstruction settings to obtain a vendor-neutral quantitative reconstruction for all systems. Volumes of interest (VOI) for the spheres were obtained by applying a 50% threshold of the sphere maximum voxel value corrected for background activity. For each sphere, the mean and maximum recovery coefficient (RCmean and RCmax) of three repeated measurements was calculated, defined as the imaged activity concentration divided by the actual activity concentration. Inter-system variations were defined as the range of RC over all systems. RC decreased with decreasing sphere volume. Inter-system variations with vendor-specific reconstructions were between 0.06 and 0.41 for RCmean depending on sphere size (maximum 118% quantification difference), and improved to 0.02-0.19 with vendor-neutral reconstructions (maximum 38% quantification difference). This study shows that eliminating sources of possible variation drastically reduces inter-system variation in quantification. This means that absolute SPECT quantification for 177Lu is feasible in a multi-center and multi-vendor setting; however, close agreement between vendors and sites is key for multi-center dosimetry and quantitative biomarker studies.


What is the Accuracy of Nuclear Imaging in the Assessment of Periprosthetic Knee Infection? A Meta-analysis.
In the assessment of possible periprosthetic knee infection, various imaging modalities are used without consensus regarding the most accurate technique. To perform a meta-analysis to compare the accuracy of various applied imaging modalities in the assessment of periprosthetic knee infection. A systematic review and meta-analysis was conducted with a comprehensive search of MEDLINE and Embase® in accordance with the PRISMA and Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) recommendations to identify clinical studies in which periprosthetic knee infection was investigated with different imaging modalities. The sensitivity and specificity of each imaging technique were determined and compared with the results of microbiologic and histologic analyses, intraoperative findings, and clinical followup of more than 6 months. A total of 23 studies, published between 1990 and 2015, were included for meta-analysis, representing 1027 diagnostic images of symptomatic knee prostheses. Quality of the included studies showed low concerns regarding external validity, whereas internal validity indicated more concerns regarding the risk of bias. The most important concerns were found in the lack of uniform criteria for the diagnosis of a periprosthetic infection and the flow and timing of the included studies. Differences among techniques were tested at a probability less than 0.05 level. Where there was slight overlap of confidence intervals for two means, it is possible for the point estimates to be statistically different from one another at a probability less than 0.05. The z-test was used to statistically analyze differences in these situations. Bone scintigraphy was less specific than all other modalities tested (56%; 95% CI, 0.47-0.64; p < 0.001), and leukocyte scintigraphy (77%; 95% CI, 0.69-0.85) was less specific than antigranulocyte scintigraphy (95%; 95% CI, 0.88-0.98; p < 0.001) or combined leukocyte and bone marrow scintigraphy (93%; 95% CI, 0.86-0.97; p < 0.001). Fluorodeoxyglucose positron emission tomography (FDG-PET) (84%; 95% CI, 0.76-0.90) was more specific than bone scintigraphy (56%; 95% CI, 0.47-0.64; p < 0.001), and less specific than antigranulocyte scintigraphy (95%; 95% CI, 0.88-0.98; p = 0.02) and combined leukocyte and bone marrow scintigraphy (93%; 95% CI, 0.86-0.97; p < 0.001). Leukocyte scintigraphy (88%; 95% CI, 0.81-0.93; p = 0.01) and antigranulocyte scintigraphy (90%; 95% CI, 0.78-0.96; p = 0.02) were more sensitive than FGD-PET (70%; 95% CI, 0.56-0.81). However, because of broad overlapping of confidence intervals, no differences in sensitivity were observed among the other modalities, including combined bone scintigraphy (93%; 95% CI, 0.85-0.98) or combined leukocyte and bone marrow scintigraphy (80%; 95% CI, 0.66-0.91; p > 0.05 for all paired comparisons). Based on current evidence, antigranulocyte scintigraphy and combined leukocyte and bone marrow scintigraphy appear to be highly specific imaging modalities in confirming periprosthetic knee infection. Bone scintigraphy was a highly sensitive imaging technique but lacks the specificity needed to differentiate among various conditions that cause painful knee prostheses. FDG-PET may not be the preferred imaging modality because it is more expensive and not more effective in confirming periprosthetic knee infection. Level III, diagnostic study.


Tonsillectomy for periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis syndrome (PFAPA).
Periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis (PFAPA) syndrome is a rare clinical syndrome of unknown cause usually identified in children. Tonsillectomy is considered a potential treatment option for this syndrome. This is an update of a Cochrane review first published in 2010. To assess the effectiveness and safety of tonsillectomy (with or without adenoidectomy) in children with PFAPA. We searched the Cochrane Ear, Nose and Throat Disorders Group Trials Register; the Cochrane Central Register of Controlled Trials (CENTRAL); PubMed; EMBASE; CINAHL; Web of Science; Cambridge Scientific Abstracts; ICTRP and additional sources for published and unpublished trials. The date of the search was 30 October 2013. Randomised controlled trials comparing tonsillectomy (with or without adenoidectomy) with non-surgical treatment in children with PFAPA. Two authors independently assessed trial quality and extracted data. We used the standard methodological procedures expected by The Cochrane Collaboration. Two trials were included with a total of 67 children randomised (65 analysed); we judged both to be at low risk of bias.One trial of 39 participants recruited children with PFAPA syndrome diagnosed according to rigid, standard criteria. The trial compared adenotonsillectomy to watchful waiting and followed up patients for 18 months. A smaller trial of 28 children applied less stringent criteria for diagnosing PFAPA and probably also included participants with alternative types of recurrent pharyngitis. This trial compared tonsillectomy alone to no treatment and followed up patients for six months.Combining the trial results suggests that patients with PFAPA experience less fever and less severe episodes after surgery compared to those receiving no surgery. The risk ratio (RR) for immediate resolution of symptoms after surgery that persisted until the end of follow-up was 4.38 (95% confidence interval (CI) 0.64 to 30.11); number needed to treat to benefit (NNTB) = 2, calculated based on an estimate that 156 in 1000 untreated children have a resolution).There was a large overall reduction in the average number of episodes over the total length of follow-up in these studies (rate ratio 0.08, 95% CI 0.05 to 0.13), reducing the average frequency of PFAPA episodes from one every two months to slightly less than one every two years. The severity, as indicated by the length of PFAPA symptoms during these episodes, was also reduced. One study reported that the average number of days per PFAPA episode was 1.7 days after receiving surgery, compared to 3.5 days in the control group. The proportion of patients requiring corticosteroids was also lower in the surgery group compared to those receiving no surgery (RR 0.58, 95% CI 0.37 to 0.92).Both trials reported that there were no complications of surgery. However, the numbers of patients randomly allocated to surgery (19 and 14 patients respectively) were too small to detect potentially important complications such as haemorrhage. Other outcomes such as quality of life, number of days with pain after surgery and absence from school were not measured or reported. The evidence for the effectiveness of tonsillectomy in children with PFAPA syndrome is derived from two small randomised controlled trials. These trials reported significant beneficial effects of surgery compared to no surgery on immediate and complete symptom resolution (NNTB = 2) and a substantial reduction in the frequency and severity (length of episode) of any further symptoms experienced. However, the evidence is of moderate quality (further research is likely to have an important impact on our confidence in the estimate of effect and may change the estimate) due to the relatively small sample sizes of the studies and some concerns about the applicability of the results. Therefore, the parents and carers of children with PFAPA syndrome must weigh the risks and consequences of surgery against the alternative of using medications. It is well established that children with PFAPA syndrome recover spontaneously and medication can be administered to try and reduce the severity of individual episodes. It is uncertain whether adenoidectomy combined with tonsillectomy adds any additional benefit to tonsillectomy alone.


Gran1: A Granulysin-Derived Peptide with Potent Activity against Intracellular Mycobacterium tuberculosis.
Granulysin is an antimicrobial peptide (AMP) expressed by human T-lymphocytes and natural killer cells. Despite a remarkably broad antimicrobial spectrum, its implementation into clinical practice has been hampered by its large size and off-target effects. To circumvent these limitations, we synthesized a 29 amino acid fragment within the putative cytolytic site of Granulysin (termed "Gran1"). We evaluated the antimicrobial activity of Gran1 against the major human pathogen Mycobacterium tuberculosis (Mtb) and a panel of clinically relevant non-tuberculous mycobacteria which are notoriously difficult to treat. Gran1 efficiently inhibited the mycobacterial proliferation in the low micro molar range. Super-resolution fluorescence microscopy and scanning electron microscopy indicated that Gran1 interacts with the surface of Mtb, causing lethal distortions of the cell wall. Importantly, Gran1 showed no off-target effects (cytokine release, chemotaxis, cell death) in primary human cells or zebrafish embryos (cytotoxicity, developmental toxicity, neurotoxicity, cardiotoxicity). Gran1 was selectively internalized by macrophages, the major host cell of Mtb, and restricted the proliferation of the pathogen. Our results demonstrate that the hypothesis-driven design of AMPs is a powerful approach for the identification of small bioactive compounds with specific antimicrobial activity. Gran1 is a promising component for the design of AMP-containing nanoparticles with selective activity and favorable pharmacokinetics to be pushed forward into experimental in vivo models of infectious diseases, most notably tuberculosis.


Bifidobacteria inhibit the inflammatory response induced by gliadins in intestinal epithelial cells via modifications of toxic peptide generation during digestion.
Celiac disease (CD) is a chronic enteropathy triggered by intake of gliadin, the toxic component of gluten. This study aims at evaluating the capacity of different Bifidobacterium strains to counteract the inflammatory effects of gliadin-derived peptides in intestinal epithelial (Caco-2) cells. A commercial extract of several gliadin (Gld) types (alpha, beta, gamma, [symbol: see text] ) was subjected to in vitro gastrointestinal digestion (pepsin at pH 3, pancreatin-bile at pH 6), inoculated or not with cell suspensions (10(8) colony forming units/ml) of either B. animalis IATA-A2, B. longum IATA-ES1, or B. bifidum IATA-ES2, in a bicameral system. The generated gliadin-derived peptides were identified by reverse phase-HPLC-ESI-MS/MS. Caco-2 cell cultures were exposed to the different gliadin peptide digestions (0.25 mg protein/ml), and the mRNA expression of nuclear factor kappa-B (NF-kappaB), tumor necrosis factor alpha (TNF-alpha), and chemokine CXCR3 receptor were analyzed by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in stimulated cells. The production of the pro-inflammatory markers NF-kappaB p50, TNF-alpha, and IL-1beta (interleukine 1beta) by Caco-2 cells was also determined by ELISA. The peptides from gliadin digestions inoculated with bifidobacteria did not exhibit the toxic amino acid sequences identified in those noninoculated (alpha/beta-Gld [158-164] and alpha/beta-Gld [122-141]). The RT-PCR analysis evidenced a down-regulation in mRNA expression of pro-inflammatory biomarkers. Consistent with these results the production of NF-kappaB, TNF-alpha, and IL-1beta was reduced (18.2-22.4%, 28.0-64.8%, and abolished, respectively) in cell cultures exposed to gliadin digestions inoculated with bifidobacteria. Therefore, bifidobacteria change the gliadin-derived peptide pattern and, thereby, attenuate their pro-inflammatory effects on Caco-2 cells.


Cortical neurodynamics of inhibitory control.
The ability to inhibit prepotent responses is critical for successful goal-directed behaviors. To investigate the neural basis of inhibitory control, we conducted a magnetoencephalography study where human participants performed the antisaccade task. Results indicated that neural oscillations in the prefrontal cortex (PFC) showed significant task modulations in preparation to suppress saccades. Before successfully inhibiting a saccade, beta-band power (18-38 Hz) in the lateral PFC and alpha-band power (10-18 Hz) in the frontal eye field (FEF) increased. Trial-by-trial prestimulus FEF alpha-band power predicted successful saccadic inhibition. Further, inhibitory control enhanced cross-frequency amplitude coupling between PFC beta-band (18-38 Hz) activity and FEF alpha-band activity, and the coupling appeared to be initiated by the PFC. Our results suggest a generalized mechanism for top-down inhibitory control: prefrontal beta-band activity initiates alpha-band activity for functional inhibition of the effector and/or sensory system.


Apoptogenic factors released from mitochondria.
When cells kill themselves, they usually do so by activating mechanisms that have evolved specifically for that purpose. These mechanisms, which are broadly conserved throughout the metazoa, involve two processes: activation in the cytosol of latent cysteine proteases (termed caspases), and disruption of mitochondrial functions. These processes are linked in a number of different ways. While active caspases can cleave proteins in the mitochondrial outer membrane, and cleave and thereby activate certain pro-apoptotic members of the Bcl-2 family, proteins released from the mitochondria can trigger caspase activation and antagonise IAP family proteins. This review will focus on the pro-apoptotic molecules that are released from the mitochondria of cells endeavouring to kill themselves. This article is part of a Special Issue entitled Mitochondria: the deadly organelle.


MK-801, but not naloxone, attenuates high-dose dextromethorphan-induced convulsive behavior: Possible involvement of the GluN2B receptor.
Dextromethorphan (DM) is a dextrorotatory isomer of levorphanol, a typical morphine-like opioid. When administered at supra-antitussive doses, DM produces psychotoxic and neurotoxic effects in humans. Although DM abuse has been well-documented, few studies have examined the effects of high-dose DM. The present study aimed to explore the effects of a single high dose of DM on mortality and seizure occurrence. After intraperitoneal administration with a high dose of DM (80mg/kg), Sprague-Dawley rats showed increased seizure occurrence and intensity. Hippocampal expression levels of N-methyl-d-aspartate (NMDA) receptor subunits (GluN1<GluN2A<GluN2B), c-Fos and pro-apoptotic factors (Bax and cleaved caspase-3) were upregulated by DM treatment; while levels of anti-apoptotic factors (Bcl-2 and Bcl-xL) were downregulated. Consistently, DM also induced ultrastructural degeneration in the hippocampus. A non-competitive NMDA receptor antagonist, MK-801, attenuated these effects of high-dose DM, whereas an opioid antagonist, naloxone, did not affect DM-induced neurotoxicity. Moreover, pretreatment with a highly specific GluN2B subunit inhibitor, traxoprodil, was selectively effective in preventing DM-induced c-Fos expression and apoptotic changes. These results suggest that high-dose DM produces convulsive behaviors by activating GluN2B/NMDA signaling that leads to pro-apoptotic changes.


Quantitative computed tomography evaluation of pulmonary disease.
Objective assessment of pulmonary disease from computed tomography (CT) examinations is desirable but difficult. When such assessments can be made, it is important that they are related to some part of the pathophysiologic process present. Herein we propose that automated volume histogram analysis can yield data that allow differentiation of normal from abnormal lung, and that the magnitude of disease will have an association with objective CT indices. Data from pulmonary CT images from 34 foxes (six uninfected controls and 28 infected with Angiostrongylus vasorum, subdivided by age and infective dose) were available. Lung tissue was segmented from surrounding tissue using an automated segmentation method. A volume histogram showing voxel frequency for each CT number in the range -1024 to -250 HU was created from the entire image stack from each fox. Using these data, the inter-quartile range and the CT number at the 95th percentile were determined. The results showed that segmentation could be readily achieved but that areas of severely diseased lung were excluded. Based on two-way analysis of variance for both the inter-quartile range and the CT number at the 95th percentile, both quantities were significantly affected by the infection status of the animal and were related to worm burden (P < 0.001). The study shows that this form of analysis is readily achieved and provides quantitative data that can be used to assess disease severity, progression, and response to treatment.


Physical, psychological, and functional comorbidities of multisymptom illness in Australian male veterans of the 1991 Gulf War.
Multisymptom illness is more prevalent in 1991 Gulf War veterans than in military comparison groups; less is known about comorbidities. The authors compared physical, psychological, and functional comorbidities in Australian male Gulf War I veterans with those in actively (non-Gulf) deployed and nondeployed military personnel by using a questionnaire and medical assessment in 2000-2002. Multisymptom illness was more common in male Gulf War veterans than in the comparison group (odds ratio (OR) = 1.80, 95% confidence interval (CI): 1.48, 2.19). Stratifying by deployment status in the comparison group made little difference in this association. Gulf War veterans with multisymptom illness had increased psychiatric disorders, including major depression (OR = 6.31, 95% CI: 4.19, 9.52) and posttraumatic stress disorder (OR = 9.77, 95% CI: 5.39, 18.59); increased unexplained chronic fatigue (OR = 13.32, 95% CI: 7.70, 23.05); and more reported functional impairment and poorer quality of life, but objective physical and laboratory outcomes were similar to those for veterans without multisymptom illness. Similar patterns were found in the comparison groups; differences across the 3 groups were statistically significant for only hospitalization, obstructive liver disease, and Epstein-Barr virus exposure. Multisymptom illness is more prevalent in Gulf War I veterans, but the pattern of comorbidities is similar for actively deployed and nondeployed military personnel.


The Effect of a Freely Available Flipped Classroom Course on Health Care Worker Patient Safety Culture: A Prospective Controlled Study.
Patient safety culture is an integral aspect of good standard of care. A good patient safety culture is believed to be a prerequisite for safe medical care. However, there is little evidence on whether general education can enhance patient safety culture. Our aim was to assess the impact of a standardized patient safety course on health care worker patient safety culture. Health care workers from Intensive Care Units (ICU) at two hospitals (A and B) in Hong Kong were recruited to compare the changes in safety culture before and after a patient safety course. The BASIC Patient Safety course was administered only to staff from Hospital A ICU. Safety culture was assessed in both units at two time points, one before and one after the course, by using the Hospital Survey on Patient Safety Culture questionnaire. Responses were coded according to the Survey User's Guide, and positive response percentages for each patient safety domain were compared to the 2012 Agency for Healthcare Research and Quality ICU sample of 36,120 respondents. We distributed 127 questionnaires across the two hospitals with an overall response rate of 74.8% (95 respondents). After the safety course, ICU A significantly improved on teamwork within hospital units (P=.008) and hospital management support for patient safety (P<.001), but decreased in the frequency of reporting mistakes compared to the initial survey (P=.006). Overall, ICU A staff showed significantly greater enhancement in positive responses in five domains than staff from ICU B. Pooled data indicated that patient safety culture was poorer in the two ICUs than the average ICU in the Agency for Healthcare Research and Quality database, both overall and in every individual domain except hospital management support for patient safety and hospital handoffs and transitions. Our study demonstrates that a structured, reproducible short course on patient safety may be associated with an enhancement in several domains in ICU patient safety culture.


Intraband absorption in self-assembled Ge-doped GaN/AlN nanowire heterostructures.
We report the observation of transverse-magnetic-polarized infrared absorption assigned to the s-p(z) intraband transition in Ge-doped GaN/AlN nanodisks (NDs) in self-assembled GaN nanowires (NWs). The s-p(z) absorption line experiences a blue shift with increasing ND Ge concentration and a red shift with increasing ND thickness. The experimental results in terms of interband and intraband spectroscopy are compared to theoretical calculations of the band diagram and electronic structure of GaN/AlN heterostructured NWs, accounting for their three-dimensional strain distribution and the presence of surface states. From the theoretical analysis, we conclude that the formation of an AlN shell during the heterostructure growth applies a uniaxial compressive strain which blue shifts the interband optical transitions but has little influence on the intraband transitions. The presence of surface states with density levels expected for m-GaN plane charge-deplete the base of the NWs but is insufficient to screen the polarization-induced internal electric field in the heterostructures. Simulations show that the free-carrier screening of the polarization-induced internal electric field in the NDs is critical to predicting the photoluminescence behavior. The intraband transitions, on the other hand, are blue-shifted due to many-body effects, namely, the exchange interaction and depolarization shift, which exceed the red shift induced by carrier screening.


Effects of mutations F61A and A62V in the fingers subdomain of HIV-1 reverse transcriptase on the translocational equilibrium.
Changes of the translocational status of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) can affect susceptibility to antiretroviral drugs. The pyrophosphate analogue phosphonoformic acid (PFA) binds specifically to and traps the pretranslocated complex of HIV-1 RT, while nucleotide-competing RT inhibitors trap the posttranslocated conformation. Here, we attempted to assess the potential role of residues in the fingers subdomain as determinants of polymerase translocation. The fingers can exist in open and closed conformations; however, the relationship between such conformational changes and the translocation status of HIV-1 RT remains elusive. We focused on substitution F61A and the neighboring A62V that is frequently associated with drug-resistance-conferring mutations. The proximity of these residues to the nucleic acid substrate suggested a possible role in translocation for these amino acid changes. We employed site-specific footprinting, binding assays, and DNA-synthesis inhibition experiments to study F61A and A62V, alone and against a background of known drug-resistance mutations. We demonstrate that F61A causes a strong bias to the posttranslocational state, while A62V shows a subtle bias toward pretranslocation regardless of the mutational background. Increases in the population of pretranslocated complexes were accompanied by increases in PFA activity, while F61A is literally resistant to PFA. Our data shed light on equilibria between pre- and posttranslocated complexes with the fingers subdomain in its open or closed conformations. We propose that a binary, pretranslocated complex in a closed conformation is stabilized with A62V and destabilized with F61A.


Lack of involvement of nucleotide excision repair gene polymorphisms in colorectal cancer.
DNA repair has an essential role in protecting the genome from damage by endogenous and environmental agents. Polymorphisms in DNA repair genes and differences in repair capacity between individuals have been widely documented. For colorectal cancer, the loss of mismatch repair gene activity is a key genetic determinant. Nucleotide excision repair (NER), recombination repair (RR) and base excision repair (BER) pathways have critical roles in protection against other cancers, and we wished to investigate their role in colorectal cancer. We have compared the frequency of polymorphisms in the NER genes, XPD, XPF, XPG, ERCC1; in the BER gene, XRCC1; and in the RR gene, XRCC3; in colorectal cancer patients and in a control group. No significant associations were found for any of the NER gene polymorphisms or for the XRCC1 polymorphism. The C allele (position 18067) of the XRCC3 gene was weakly but significantly associated with colorectal cancer (odds ratio 1.52, 95% confidence interval 1.04-2.22, P=0.03). For all patients who were heterozygous for any of the repair genes studied, tumour tissue was investigated for loss of heterozygosity (LOH). Only one example of LOH was found for all the genes examined. From the association and LOH data, we conclude that these genes do not have an important role in protection against colorectal carcinogenesis.


Respiratory effects of amyotrophic lateral sclerosis: problems and solutions.
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease. Most patients die from respiratory complications. Fortunately, there are a growing number of treatment options that can improve both survival and quality of life for patients with ALS. This review discusses the respiratory evaluation and treatment of patients with ALS, about which a great deal is known. It also includes material on related problems, such as speech and swallowing difficulties and end-of-life care.


Effectiveness of MB-102, a novel fluorescent tracer agent, for conducting ocular angiography in dogs.
To evaluate the effectiveness of a novel fluorescence tracer agent, MB-102, for conducting ocular angiography in dogs. 10 ophthalmologically normal dogs (2 to 4 years old) and 10 dogs with retinal degeneration or primary open-angle glaucoma (< 6 years old). While anesthetized, all dogs received sodium fluorescein (20 mg/kg, IV) or MB-102 (20 or 40 mg/kg, IV) first and then the other dye in a second treatment session 2 days later in a randomized crossover design. Anterior fluorescence angiography was performed on one eye and posterior fluorescence angiography on the other. Imaging was performed with a full-spectrum camera and camera adaptor system. Filter sets that were tailored to match the excitation and emission characteristics of each angiographic fluorescent agent were used. All phases and phase intervals during anterior and posterior segment angiography were identified, regardless of the dye used. However, agent fluorescence and visualization of the iridal blood vessels were hindered in some dogs, irrespective of agent, owing to the degree of iridal pigmentation present. No significant difference was noted between the 2 dyes in any phase or phase interval, and slight improvement in image contrast was observed with MB-102 during the venous phases owing to a reduction of vessel wall staining in both normal and diseased eyes. Results suggested that MB-102 would be useful for conducting ocular angiography in dogs.


Novel methodology to characterize electromagnetic exposure of the brain.
Due to the greatly non-uniform field distribution induced in brain tissues by radio frequency electromagnetic sources, the exposure of anatomical and functional regions of the brain may be a key issue in interpreting laboratory findings and epidemiological studies concerning endpoints related to the central nervous system. This paper introduces the Talairach atlas in characterization of the electromagnetic exposure of the brain. A hierarchical labeling scheme is mapped onto high-resolution human models. This procedure is fully automatic and allows identification of over a thousand different sites all over the brain. The electromagnetic absorption can then be extracted and interpreted in every region or combination of regions in the brain, depending on the characterization goals. The application examples show how this methodology enhances the dosimetry assessment of the brain based on results obtained by either finite difference time domain simulations or measurements delivered by test compliance dosimetry systems. Applications include, among others, the detailed dosimetric analysis of the exposure of the brain during cell phone use, improved design of exposure setups for human studies or medical diagnostic and therapeutic devices using electromagnetic fields or ultrasound.


Target identification by image analysis.
Covering: 1997 to the end of 2015Each biologically active compound induces phenotypic changes in target cells that are characteristic for its mode of action. These phenotypic alterations can be directly observed under the microscope or made visible by labelling structural elements or selected proteins of the cells with dyes. A comparison of the cellular phenotype induced by a compound of interest with the phenotypes of reference compounds with known cellular targets allows predicting its mode of action. While this approach has been successfully applied to the characterization of natural products based on a visual inspection of images, recent studies used automated microscopy and analysis software to increase speed and to reduce subjective interpretation. In this review, we give a general outline of the workflow for manual and automated image analysis, and we highlight natural products whose bacterial and eucaryotic targets could be identified through such approaches.


Formation of [20R]-dihydrodigoxin from digoxin in humans.
The objective of this research was to determine the stereochemical identity of dihydrodigoxin (DHD3), a metabolite of digoxin excreted in urine after digoxin administration in man. Separation of the individual epimers in reference DHD3 was effected by a chemical derivatization-HPLC procedure. Comparison of individual derivatized epimers of DHD3 with the known 20R and 20S epimers of derivatized dihydrodigoxigenin using chromatographic data and NMR spectroscopy permitted identification of the 20R and 20S epimers in reference DHD3. Material corresponding chromatographically to R-DHD3 was isolated from urine of a volunteer taking oral digoxin daily. The NMR spectrum of the chromatographically pure, derivatized urinary isolate was identical to the NMR spectrum of derivatized R-DHD3. Urine samples from 20 patients on chronic digoxin therapy and from two volunteers were surveyed for chromatographic evidence of R- or S-DHD3 using HPLC of a fluorescent derivative as well as HPLC of a UV-absorbing derivative. The more sensitive fluorescence procedure yielded evidence for R-DHD3 in eight patients and both volunteers. There was a sufficient concentration in four patients and both volunteers for independent evidence of R-DHD3 using the UV-absorbing derivative. Chromatographic evidence (fluorescent derivative) for S-DHD3 was found in urine from three patients. However, this evidence for S-DHD3 was demonstrated to be an artifact by the absence of the peak expected for S-DHD3 in the chromatogram of the UV-absorbing derivative as well as by the inability of fecal incubates to convert digoxin to S-DHD3. The results of the investigation indicate that the DHD3 metabolite formed in humans is the 20R epimer.


Effect of standardized cranberry extract on the activity and expression of selected biotransformation enzymes in rat liver and intestine.
The use of dietary supplements containing cranberry extract is a common way to prevent urinary tract infections. As consumption of these supplements containing a mixture of concentrated anthocyanins and proanthocyanidins has increased, interest in their possible interactions with drug-metabolizing enzymes has grown. In this in vivo study, rats were treated with a standardized cranberry extract (CystiCran®) obtained from Vaccinium macrocarpon in two dosage schemes (14 days, 0.5 mg of proanthocyanidins/kg/day; 1 day, 1.5 mg of proanthocyanidins/kg/day). The aim of this study was to evaluate the effect of anthocyanins and proanthocyanidins contained in this extract on the activity and expression of intestinal and hepatic biotransformation enzymes: cytochrome P450 (CYP1A1, CYP1A2, CYP2B and CYP3A), carbonyl reductase 1 (CBR1), glutathione-S-transferase (GST) and UDP-glucuronosyl transferase (UGT). Administration of cranberry extract led to moderate increases in the activities of hepatic CYP3A (by 34%), CYP1A1 (by 38%), UGT (by 40%), CBR1 (by 17%) and GST (by 13%), while activities of these enzymes in the small intestine were unchanged. No changes in the relative amounts of these proteins were found. Taken together, the interactions of cranberry extract with simultaneously administered drugs seem not to be serious.


Comparative evaluation of prophylactic use of pregabalin, gabapentin and diclofenac sodium for prevention of succinylcholine-induced myalgia: a randomized, double-blinded study.
Succinylcholine a depolarizing muscle relaxant with rapid onset, predictable course and short duration of action is associated with myalgia. The aim of this study is to evaluate the efficacy of pregabalin, gabapentin and diclofenac on the incidence and severity of succinylcholine-induced myalgia. Tertiary Care Teaching Hospital. A total of 120 patients undergoing laparoscopic cholecystectomy were randomly assigned into three groups: Pregabalin group received 150 mg of pregabalin, gabapentin group received 600 mg of gabapentin and diclofenac group received 100 mg of diclofenac sodium orally 2 h prior to surgery. Anesthesia was induced with fentanyl 3 μg/kg, propofol 2-2.5 mg/kg and succinylcholine 1.5 mg/kg and was maintained with oxygen with sevoflurane in the air and intermittent vecuronium bromide. A blinded observer recorded post-operative pain scores on visual analog scale at different time intervals and myalgia at 24 h. Post-operative pain relief was provided with fentanyl based patient-controlled analgesia. Fentanyl consumption in 24 h was recorded as a primary outcome. Patients' characteristics and total fentanyl consumption were compared using one-way ANOVA followed by post-hoc test. Pain score was compared amongst the groups using Kruskal Wallis test. The myalgia occurred in 15, 14 and 13 patients in pregabalin, gabapentin and diclofenac sodium group respectively (P > 0.85). Patients in diclofenac group had significantly higher fentanyl consumption (674.85 ± 115.58 μg) compared with pregabalin group (601.87 ± 129.57 μg) (95% confidence interval [CI] = 34.8-120.7) and gabapentin group (612.29 ± 105.12 μg) (95% CI = 14.9-170.5). However, there was no significant difference in fentanyl consumption between pregabalin and gabapentin groups (95% CI = -34.8-120.7). There was a significant difference in visual analog score at time points 12, 18 and 24 h among the study groups. Pre-treatment with pregabalin, gabapentin and diclofenac had equal efficacy in reducing the incidence and severity of succinylcholine-induced myalgia. However, pre-treatment with pregabalin and gabapentin decreased post-operative pain scores and fentanyl consumption.