Optimizing tamoxifen-inducible Cre/loxp system to reduce tamoxifen effect on bone turnover in long bones of young mice.
For tamoxifen-dependent Cre recombinase, also known as CreER recombinase, tamoxifen (TAM) is used to activate the Cre to generate time- and tissue-specific mouse mutants. TAM is a potent CreER system inducer; however, TAM is also an active selective estrogen receptor modulator (SERM) that can influence bone homeostasis. The purpose of this study was to optimize the TAM dose for Cre recombinase activation while minimizing the effects of TAM on bone turnover in young growing mice. To evaluate the effects of TAM on bone turnover and bone mass, 1-month-old wild-type male and female mice were intraperitoneally injected with TAM at 0, 1, 10 or 100mg/kg/day for four consecutive days, or 100, 300 mg/kg/day for one day. The distal femurs were analyzed one month after the last TAM injection by microCT, mechanical test, and surface-based bone histomorphometry. Similar doses of TAM were used in Col1 (2.3 kb)-CreERT2; mT/mG reporter male mice to evaluate the dose-dependent efficacy of Cre-ER activation in bone tissue. A TAM dose of 100 mg/kg × 4 days significantly increased trabecular bone volume/total volume (BV/TV) of the distal femur, femur length, bone strength, and serum bone turnover markers compared to the 0mg control group. In contrast, TAM doses ≤ 10 mg/kg did not significantly change any of these parameters compared to the 0mg group, although a higher bone strength was observed in the 10mg group. Surface-based histomorphometry revealed that the 100mg/kg dose of TAM dose significantly increased trabecular bone formation and decreased periosteal bone formation at 1-week post-TAM treatment. Using the reporter mouse model Col1-CreERT2; mT/mG, we found that 10mg/kg TAM induced Col1-CreERT2 activity in bone at a comparable level to the 100mg/kg dose. TAM treatment at 100mg/kg/day × 4 days significantly affects bone homeostasis, resulting in an anabolic bone effect on trabecular bone in 1-month-old male mice. However, a lower dose of TAM at 10 mg/kg/day × 4 days can yield similar Col1-CreERT2 induction efficacy with minimum effects on bone turnover in young male mice.


Critical nucleotides in the interaction of CatR with the pheBA promoter: conservation of the CatR-mediated regulation mechanisms between the pheBA and catBCA operons.
The promoter of the plasmid-borne pheBA genes encoding enzymes for phenol degradation resembles the catBCA promoter and is activated by CatR, the regulator of the chromosomally encoded catechol-degradative catBCA genes in Pseudomonas putida. In this study, site-directed mutagenesis of the pheBA promoter region was performed. The interrupted inverted repeat sequence of the CatR recognition binding site (RBS) of the pheBA promoter is highly homologous to that of the catBCA promoter. However, the RBS was shown not to be the sole important feature for high-affinity binding of CatR to this site. Mutagenesis of the activation binding site (ABS) of CatR, which overlaps the -35 hexamer sequence TTGGAT of the promoter, revealed that the two G nucleotides in this sequence are important for promoter activity but not for CatR binding. All other substitutions made in the ABS negatively affected both the promoter activity and CatR binding. The spacer sequence of the pheBA and catBCA promoters between the -10 and -35 hexamers is 19 bp, which is longer than optimal. However, reducing the spacer region of the pheBA promoter was not sufficient for CatR-independent promoter activation. An internal binding site (IBS) for CatR is located downstream of the transcriptional start site of the catBCA genes and it negatively regulates the operon. A similar IBS was identified in the case of the pheBA operon and tested for its functionality. The results indicate a conservation of CatR-mediated regulation mechanisms between the pheBA promoter and the catBCA promoter. This universal mechanism of CatR-mediated transcriptional activation could be of great importance in enabling catechol-degrading bacteria to expand their substrate range via horizontal transfer of the phenol degradative genes.


Molecular analysis of human endometrium: short-term tibolone signaling differs significantly from estrogen and estrogen + progestagen signaling.
Tibolone, a tissue-selective compound with a combination of estrogenic, progestagenic, and androgenic properties, is used as an alternative for estrogen or estrogen plus progesterone hormone therapy for the treatment of symptoms associated with menopause and osteoporosis. The current study compares the endometrial gene expression profiles after short-term (21 days) treatment with tibolone to the profiles after treatment with estradiol-only (E(2)) and E(2) + medroxyprogesterone acetate (E(2) + MPA) in healthy postmenopausal women undergoing hysterectomy for endometrial prolapse. The impact of E(2) treatment on endometrial gene expression (799 genes) was much higher than the effect of tibolone (173 genes) or E(2) + MPA treatment (174 genes). Furthermore, endometrial gene expression profiles after tibolone treatment show a weak similarity to the profiles after E(2) treatment (overlap 72 genes) and even less profile similarity to E(2) + MPA treatment (overlap 17 genes). Interestingly, 95 tibolone-specific genes were identified. Translation of profile similarity into biological processes and pathways showed that ER-mediated downstream processes, such as cell cycle and cell proliferation, are not affected by E2 + MPA, slightly by tibolone, but are significantly affected by E(2). In conclusion, tibolone treatment results in a tibolone-specific gene expression profile in the human endometrium, which shares only limited resemblance to E(2) and even less resemblance to E2 + MPA induced profiles.


Determining whether a class of random graphs is consistent with an observed contact network.
We demonstrate a general method to analyze the sensitivity of attack rate in a network model of infectious disease epidemiology to the structure of the network. We use Moore and Shannon's "network reliability" statistic to measure the epidemic potential of a network. A number of networks are generated using exponential random graph models based on the properties of the contact network structure of one of the Add Health surveys. The expected number of infections on the original Add Health network is significantly different from that on any of the models derived from it. Because individual-level transmissibility and network structure are not separately identifiable parameters given population-level attack rate data it is possible to re-calibrate the transmissibility to fix this difference. However, the temporal behavior of the outbreak remains significantly different. Hence any estimates of the effectiveness of time dependent interventions on one network are unlikely to generalize to the other. Moreover, we show that in one case even a small perturbation to the network spoils the re-calibration. Unfortunately, the set of sufficient statistics for specifying a contact network model is not yet known. Until it is, estimates of the outcome of a dynamical process on a particular network obtained from simulations on a different network are not reliable.


A common haplotype of the TNF receptor 2 gene modulates endotoxin tolerance.
Endotoxin tolerance is characterized by the suppression of further TNF release upon recurrent exposure to LPS. This phenomenon is proposed to act as a homeostatic mechanism preventing uncontrolled cytokine release such as that observed in bacterial sepsis. The regulatory mechanisms and interindividual variation of endotoxin tolerance induction in man remain poorly characterized. In this paper, we describe a genetic association study of variation in endotoxin tolerance among healthy individuals. We identify a common promoter haplotype in TNFRSF1B (encoding TNFR2) to be strongly associated with reduced tolerance to LPS (p = 5.82 × 10(-6)). This identified haplotype is associated with increased expression of TNFR2 (p = 4.9 × 10(-5)), and we find basal expression of TNFR2, irrespective of genotype and unlike TNFR1, is associated with secondary TNF release (p < 0.0001). Functional studies demonstrate a positive-feedback loop via TNFR2 of LPS-induced TNF release, confirming this previously unrecognized role for TNFR2 in the modulation of LPS response.


Assembly and extension of yeast artificial chromosomes to build up a large locus.
For the assembly of a large human locus, overlapping regions on yeast artificial chromosomes (YACs) and cosmids were linked up using their regions of homology. By site-specific recombination a YAC of 410 kb was created accommodating the major part of the human lambda light chain locus in authentic configuration with 28 variable (V) genes, all joining (J) segments, all constant (C) genes and the downstream enhancer. A contiguous region was first constructed from three overlapping cosmids. Each of these was linearized at unique sites in the vectors and YAC arms were ligated to the 5' and 3' ends. After cells of Saccharomyces cerevisiae were transformed with the three cosmids, YACs of 120 kb were obtained which contained the reassembled 3' J-C region in authentic configuration. The assembled YAC was further extended by mitotic recombination with a YAC containing a 280-kb region of the C-proximal part of the V gene cluster with a 15-kb 3' overlap. For this, a simple three-way selection procedure was developed involving the integration of different selectable marker genes at specific sites in the left and right YAC arms. Rare recombination events between two overlapping YACs could be identified in yeast clones able to grow in lysine- and adenine-deficient medium in the presence of 5-fluoro-orotic acid which is toxic for yeast cells containing a YAC with a functional URA3 gene. This approach made it possible to assemble and extend large YACs from an unlimited number of smaller overlapping YACs by positive-negative selection.


Systematic determination of order parameters for chain dynamics using diffusion maps.
We employ the diffusion map approach as a nonlinear dimensionality reduction technique to extract a dynamically relevant, low-dimensional description of n-alkane chains in the ideal-gas phase and in aqueous solution. In the case of C8 we find the dynamics to be governed by torsional motions. For C16 and C24 we extract three global order parameters with which we characterize the fundamental dynamics, and determine that the low free-energy pathway of globular collapse proceeds by a "kink and slide" mechanism, whereby a bend near the end of the linear chain migrates toward the middle to form a hairpin and, ultimately, a coiled helix. The low-dimensional representation is subtly perturbed in the solvated phase relative to the ideal gas, and its geometric structure is conserved between C16 and C24. The methodology is directly extensible to biomolecular self-assembly processes, such as protein folding.


Effect of temporal lakes on avifaunal composition at the Southeast of Isthmus of Tehuantepec, Oaxaca, Mexico.
Oaxaca hosts one of the greatest biodiversity in México, occupying first place in avian diversity compared to other regions of the country. However, the area is undergoing serious problems such as high defor- estation rates, soil erosion and over exploitation and extinction of species. These factors have all contributed to the current loss of biodiversity. Also, biological inventories are still incomplete. One of the least explored sites is the semiarid zone of Tehuantepec isthmus, around the locality of Santa Maria del Mar, Oaxaca, México. The area includes floodable grasslands, mangrove areas and dry forest, providing a range of potential habitats for different species. Our objective was to evaluate the effect of temporal lakes on spatial and temporal composition of the avifauna in Santa Maria del Mar, in order to generate information regarding this group within the region and the state, and to understand the importance of flood areas for resident and transitory birds. We conducted 12 avifauna surveys between July 2006 and June 2008, and established two transects of 2km length in each of four habitat types (beach, grassland, dry forest, and mangrove). We found a total of 75 species, corresponding to 16 orders and 30 families. Within an area of 26 km2, we significantly found 10.1% of the total number of bird species recorded for the entire state, and 6.6% of the total reported in Mexico. The families most repre- sented were: Ardeidae, Laridae and Scolopacidae. Over the entire study period, dry forest was the most diverse habitat; followed by mangrove, grassland and the beach. Of all the species recorded, 38.6% were found at the edge or in the temporal lakes. We found a significant difference in species composition between seasons in the grassland, but no difference in the other habitats. Our results showed a significant effect of temporary lakes on avian diversity during the wet season; it also demonstrated the importance of grassland conservation given its relevance for presence of waterbirds. We suggested a continued and more intense monitoring; and proposed the designation of the area as an Important Conservation Bird Area (AICA).


Automated discrimination of psychotropic drugs in mice via computer vision-based analysis.
We developed an inexpensive computer vision-based method utilizing an algorithm which differentiates drug-induced behavioral alterations. The mice were observed in an open-field arena and their activity was recorded for 100 min. For each animal the first 50 min of observation were regarded as the drug-free period. Each animal was exposed to only one drug and they were injected (i.p.) with either amphetamine or cocaine as the stimulant drugs or morphine or diazepam as the inhibitory agents. The software divided the arena into virtual grids and calculated the number of visits (sojourn counts) to the grids and instantaneous speeds within these grids by analyzing video data. These spatial distributions of sojourn counts and instantaneous speeds were used to construct feature vectors which were fed to the classifier algorithms for the final step of matching the animals and the drugs. The software decided which of the animals were drug-treated at a rate of 96%. The algorithm achieved 92% accuracy in sorting the data according to the increased or decreased activity and then determined which drug was delivered. The method differentiated the type of psychostimulant or inhibitory drugs with a success ratio of 70% and 80%, respectively. This method provides a new way to automatically evaluate and classify drug-induced behaviors in mice.


Need for Two-Year Patient-Reported Outcomes Score for Lumbar Spine Surgery Is Procedure-Specific: Analysis From a Prospective Longitudinal Spine Registry.
Retrospective analysis of prospectively collected data. The aim of this study was to determine whether 1-year patient-reported outcomes (PROs) can accurately assess effective care for patients undergoing surgery for degenerative lumbar spine disease. Prospective longitudinal PROs registries provide a means to accurately assess outcomes and determine the relative effectiveness of various spine treatments. Obtaining long-term PROs can be costly and challenging. Patients enrolled into a prospective registry who underwent lumbar spine surgery for degenerative disease were included. Baseline, 1-year, and 2-year Oswestry Disability Index (ODI) scores were captured. Previously published minimum clinically important difference (MCID) for ODI (14.9) was used. Multivariable linear regression model was created to derive model-estimated 2-year ODI scores. Absolute differences between 1-year and 2-year ODI were compared to absolute differences between 2-year and model-estimated 2-year ODI. Concordance rates in achieving MCID at 1-year and 2-year and predictive values were calculated. A total of 868 patients were analyzed. One-year ODI scores differed from 2-year scores by an absolute difference of 9.7 ± 8.9 points and predictive model-estimated 2-year scores differed from actual 2-year scores by 8.8 ± 7.3 points. The model-estimated 2-year ODI was significantly different than actual 1-year ODI in assessing actual 2-year ODI for all procedures (P = 0.001) except for primary (P = 0.932) and revision microdiscectomy (P = 0.978) and primary laminectomy (P = 0.267). The discordance rates of achieving or not achieving MCID for ODI ranged from 8% to 27%. Concordance rate was about 90% for primary and revision microdiscectomy. The positive and negative predictive value of 1-year ODI to predict 2-year ODI was 83% and 67% for all procedures and 92% and 67% for primary and 100% and 86% for revision microdiscectomy respectively. One-year disability outcomes can potentially estimate 2-year outcomes for patient populations, but cannot reliably predict 2-year outcomes for individual patients, except for patients undergoing primary and revision microdiscectomy. 4.


A peptide derived from the intercellular adhesion molecule-2 regulates the avidity of the leukocyte integrins CD11b/CD18 and CD11c/CD18.
beta 2 integrin (CD11a,b,c/CD18)-mediated cell adhesion is required for many leukocyte functions. Under normal circumstances, the integrins are nonadhesive, and become adhesive for their cell surface ligands, the intercellular adhesion molecules (ICAMs), or soluble ligands such as fibrinogen and iC3b, when leukocytes are activated. Recently, we defined a peptide derived from ICAM-2, which specifically binds to purified CD11a/CD18. Furthermore, this peptide strongly induces T cell aggregation mainly mediated by CD11a/CD18-ICAM-1 interaction, and natural killer cell cytotoxicity. In the present study, we show that the same ICAM-2 peptide also avidly binds to purified CD11b/CD18, but not to CD11c/CD18. This binding can be blocked by the CD11b antibody OKM10. The peptide strongly stimulates CD11b/CD18-ICAM-1-mediated cell aggregations of the monocytic cell lines THP-1 and U937. The aggregations are energy and divalent cation-dependent. The ICAM-2 peptide also induces CD11b/CD18 and CD11c/CD18-mediated binding of THP-1 cells to fibrinogen and iC3b coated on plastic. These findings indicate that in addition to induction of CD11a/CD18-mediated cell adhesion, the ICAM-2 peptide may also serve as a "trigger" for high avidity ligand binding of other beta 2 integrins.


Insights into the molecular regulation of monolignol-derived product biosynthesis in the growing hemp hypocotyl.
Lignin and lignans are both derived from the monolignol pathway. Despite the similarity of their building blocks, they fulfil different functions in planta. Lignin strengthens the tissues of the plant, while lignans are involved in plant defence and growth regulation. Their biosyntheses are tuned both spatially and temporally to suit the development of the plant (water conduction, reaction to stresses). We propose to study the general molecular events related to monolignol-derived product biosynthesis, especially lignin. It was previously shown that the growing hemp hypocotyl (between 6 and 20 days after sowing) is a valid system to study secondary growth and the molecular events accompanying lignification. The present work confirms the validity of this system, by using it to study the regulation of lignin and lignan biosynthesis. Microscopic observations, lignin analysis, proteomics, together with in situ laccase and peroxidase activity assays were carried out to understand the dynamics of lignin synthesis during the development of the hemp hypocotyl. Based on phylogenetic analysis and targeted gene expression, we suggest a role for the hemp dirigent and dirigent-like proteins in lignan biosynthesis. The transdisciplinary approach adopted resulted in the gene- and protein-level quantification of the main enzymes involved in the biosynthesis of monolignols and their oxidative coupling (laccases and class III peroxidases), in lignin deposition (dirigent-like proteins) and in the determination of the stereoconformation of lignans (dirigent proteins). Our work sheds light on how, in the growing hemp hypocotyl, the provision of the precursors needed to synthesize the aromatic biomolecules lignin and lignans is regulated at the transcriptional and proteomic level.


Interleukin-1, tumor necrosis factor-alpha, and transforming growth factor-beta 1 and integrative meniscal repair: influences on meniscal cell proliferation and migration.
Interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) are up-regulated in injured and osteoarthritic knee joints. IL-1 and TNF-α inhibit integrative meniscal repair; however, the mechanisms by which this inhibition occurs are not fully understood. Transforming growth factor-β1 (TGF-β1) increases meniscal cell proliferation and accumulation, and enhances integrative meniscal repair. An improved understanding of the mechanisms modulating meniscal cell proliferation and migration will help to improve approaches for enhancing intrinsic or tissue-engineered repair of the meniscus. The goal of this study was to examine the hypothesis that IL-1 and TNF-α suppress, while TGF-β1 enhances, cellular proliferation and migration in cell and tissue models of meniscal repair. A micro-wound assay was used to assess meniscal cell migration and proliferation in response to the following treatments for 0, 24, or 48 hours: 0 to 10 ng/mL IL-1, TNF-α, or TGF-β1, in the presence or absence of 10% serum. Proliferated and total cells were fluorescently labeled and imaged using confocal laser scanning microscopy and the number of proliferated, migrated, and total cells was determined in the micro-wound and edges of each image. Meniscal cell proliferation was also assessed throughout meniscal repair model explants treated with 0 or 10 ng/mL IL-1, TNF-α, or TGF-β1 for 14 days. At the end of the culture period, biomechanical testing and histological analyses were also performed. Statistical differences were assessed using an ANOVA and Newman-Keuls post hoc test. IL-1 and TNF-α decreased cell proliferation in both cell and tissue models of meniscal repair. In the presence of serum, TGF-β1 increased outer zone cell proliferation in the micro-wound and in the cross section of meniscal repair model explants. Both IL-1 and TNF-α decreased the integrative shear strength of repair and extracellular matrix deposition in the meniscal repair model system, while TGF-β1 had no effect on either measure. Meniscal cell proliferation in vivo may be diminished following joint injury due to the up-regulation of inflammatory cytokines, thereby limiting native cellular repair of meniscal lesions. Therefore, therapies that can promote meniscal cell proliferation have promise to enhance meniscal repair and improve tissue engineering strategies.


Exercise-induced muscle damage and the repeated bout effect: evidence for cross transfer.
We examined whether a prior bout of eccentric exercise in the elbow flexors provided protection against exercise-induced muscle damage in the contralateral arm. Fifteen males (age 22.7 ± 2.1 years; height 178.6 ± 6.8 cm, mass 75.8 ± 9.3 kg) were randomly assigned to two groups who performed two bouts of 60 eccentric contractions (30°/s) separated by 2 weeks: ipsilateral (n = 7, both bouts performed in the same arm), contralateral (n = 8, one bout performed in each arm). Strength, muscle soreness and resting arm angle (RAA) were measured at baseline and at 1, 24 and 48 h post exercise. Surface electromyography was recorded during both bouts of exercise. The degree of strength loss was attenuated (p < 0.05) in the ipsilateral group after the second bout of eccentric exercise (-22 cf. -3% for bout 1 and 2 at 24 h, respectively). Strength loss following eccentric exercise was also attenuated (p < 0.05) at 24 h in the contralateral group (-30 cf. 13% for bout 1 and 2, respectively). Muscle soreness (≈34 cf 19 mm) and change in RAA (≈5 cf. 3%) were also lower following the second bout of eccentric exercise (p < 0.05), although there was no difference in the overall change in these values between groups. Median frequency (MF) was decreased by 31% between bouts, with no difference between groups. Data support observations that the repeated bout effect transfers to the opposite (untrained) limb. The similar reduction in MF between bouts for the two groups provides evidence for a centrally mediated, neural adaptation.


Opinions of clinical veterinarians at a US veterinary teaching hospital regarding antimicrobial use and antimicrobial-resistant infections.
To determine opinions of faculty members with clinical appointments, clinical veterinarians, residents, and interns at a US veterinary teaching hospital regarding antimicrobial use and antimicrobial-resistant infections. Cross-sectional survey. 71 veterinarians. An online questionnaire was sent to all veterinarians with clinical service responsibilities at the North Carolina State University veterinary teaching hospital (n = 167). The survey included 23 questions regarding demographic information, educational experiences, current prescribing practices, and personal opinions related to antimicrobial selection, antimicrobial use, restrictions on antimicrobial use, and antimicrobial resistance. Of the 167 veterinarians eligible to participate, 71 (43%) responded. When respondents were asked to rate their level of concern (very concerned = 1; not concerned = 5) about antimicrobial-resistant infections, most (41/70 [59%]) assigned a score of 1, with mean score for all respondents being 1.5. Most survey participants rated their immediate colleagues (mean score, 1.9) as more concerned than other veterinary medical professionals (mean score, 2.3) and their clients (mean score, 3.4). Fifty-nine of 67 (88%) respondents felt that antimicrobials were overprescribed at the hospital, and 32 of 69 (46%) respondents felt uncomfortable prescribing at least one class of antimicrobials (eg, carbapenems or glycopeptides) because of public health concerns. Findings indicated that veterinarians at this teaching hospital were concerned about antimicrobial resistance, thought antimicrobials were overprescribed, and supported restricting use of certain antimicrobial classes in companion animals. Findings may be useful in educating future veterinarians and altering prescribing habits and antimicrobial distribution systems in veterinary hospitals.


Reliability evaluation of thermophysical properties from first-principles calculations.
Thermophysical properties, such as heat capacity, bulk modulus and thermal expansion, are of great importance for many technological applications and are traditionally determined experimentally. With the rapid development of computational methods, however, first-principles computed temperature-dependent data are nowadays accessible. We evaluate various computational realizations of such data in comparison to the experimental scatter. The work is focussed on the impact of different first-principles codes (QUANTUM ESPRESSO and VASP), pseudopotentials (ultrasoft and projector augmented wave) as well as phonon determination methods (linear response and direct force constant method) on these properties. Based on the analysis of data for two pure elements, Cr and Ni, consequences for the reliability of temperature-dependent first-principles data in computational thermodynamics are discussed.


Detection of autonomic sympathetic dysfunction in diabetic patients. A study using laser Doppler imaging.
To study signs of the disturbed reflex autonomic sympathetic nerve function in type 1 and type 2 diabetic patients. Measurements were made on 15 type 1 (duration 13-32 years) and on 50 recently diagnosed type 2 diabetic patients (duration 3-4 years). The vasoconstrictor responses in the distal phalanx of the middle finger (locally heated to 40 degrees C) to the cooling of the contralateral arm were measured using Laser Doppler Imaging (LDI). A vasoconstriction index (VAC) was calculated taking age into account and was compared with reference values obtained in 80 control subjects. The diabetic patients were also studied with deep-breathing tests (i.e., the heart-rate variation expressed as the expiration-to-inspiration [E/I] ratio, a test of parasympathetic nerve function). The vasoconstrictor responses to indirect cooling (VAC) were significantly reduced in the fingers of the diabetic patients, both type 2 (0.77 +/- 0.02 V; P < 0.01) and type 1 (0.83 +/- 0.04 V; P < 0.001), compared with the healthy control subjects (0.65 +/- 0.01); the age-corrected VAC (VACz) was slightly more impaired in type 1 than in type 2 diabetic patients. The frequency of an abnormal VACz corresponded well to the frequency of an abnormal E/I ratio in type 1 diabetic patients (approximately 50%), whereas the frequency of an abnormal VACz was significantly higher than an abnormal E/I ratio among type 2 diabetic patients (11/50 vs. 4/50; P < 0.05). Both type 1 and type 2 diabetic patients have impaired cutaneous blood flow regulation. The VAC index seems to be a promising tool for detection of subclinical changes in autonomic sympathetic function.


Post-traumatic head injury pituitary dysfunction.
Partial or complete pituitary dysfunction affects 33-50% of all traumatic brain injury (TBI) survivors and is a significant contributor to the overall disability burden. The hypophyseal vessels are anatomically vulnerable to shearing injuries, raised intracranial pressure and anterior base of skull fractures, and pituitary ischaemia or haemorrhage is a common finding at autopsy. Post-traumatic hypopituitarism (PTHP) can affect all grades of severity of injury and is often difficult to diagnose, as its features largely overlap with common post-concussive symptoms. PTHP has a wide range of manifestations, including fatigue, myopathy, cognitive difficulties, depression, behavioural changes or life-threatening complications such as sodium dysregulation and adrenal crisis. In some instances, mild PTHP can recover, at least partially, but cases of late onset are also known. At present, there is no consensus on whether all TBI patients should be screened (including mild TBI) and at what time points, given that neuroendocrine tests in the acute phase are simply likely to reflect a non-specific trauma response rather than true pituitary damage and that the time course of PTHP is unclear. A full investigation of the hypothalamic-pituitary axis requires specialized neuroendocrine assessment, including stimulation tests, as random hormone levels can be misleading in this context. Given the high incidence of TBI, this may have significant resource implications for Endocrinology services but, on the other hand, patients with PTHP may receive suboptimal rehabilitation unless the underlying hormone deficiency is identified and treated.


Production of giant mouse oocyte nucleoli and assessment of their protein content.
Compared with advanced developmental stage embryos and somatic cells, fully grown mammalian oocytes contain specific nucleolus-like structures (NPB - nucleolus precursor bodies). It is commonly accepted that they serve as a store of material(s) from which typical nucleoli are gradually formed. Whilst nucleoli from somatic cells can be collected relatively easily for further biochemical analyses, a sufficient number of oocyte nucleoli is very difficult to obtain. We have found that isolated oocytes nucleoli fuse very efficiently when contact is established between them. Thus, well visible giant nucleoli can be obtained, relatively easily handled and then used for further biochemical analyses. With the use of colloidal gold staining, we estimated that a single fully grown mouse oocyte nucleolus contains approximately 1.6 ng of protein. We do believe that this approach will accelerate further research aiming at analyzing the composition of oocyte nucleoli in more detail.


Classical and Quantum Shortcuts to Adiabaticity in a Tilted Piston.
Adiabatic quantum state evolution can be accelerated through a variety of shortcuts to adiabaticity. In one approach, a counterdiabatic quantum Hamiltonian, ĤCD, is constructed to suppress nonadiabatic excitations. In the analogous classical problem, a counterdiabatic classical Hamiltonian, HCD, ensures that the classical action remains constant even under rapid driving. Both the quantum and classical versions of this problem have been solved for the special case of scale-invariant driving, characterized by linear expansions, contractions, or translations of the system. Here we investigate an example of a non-scale-invariant system, a tilted piston. We solve exactly for the classical counterdiabatic Hamiltonian, HCD(q, p, t), which we then quantize to obtain a Hermitian operator, ĤCD(t). Using numerical simulations, we find that ĤCD effectively suppresses nonadiabatic excitations under rapid driving. These results offer a proof of principle, beyond the special case of scale-invariant driving, that quantum shortcuts to adiabaticity can successfully be constructed from their classical counterparts.


Clinical Sequencing Contributes to a BRCA-Associated Cancer Rediagnosis That Guides an Effective Therapeutic Course.
Cancer is currently classified and treated using an approach based on tissue of origin. Ambiguous or incorrect diagnoses, however, are common and often go unnoticed. Clinical cancer sequencing can provide diagnostic precision, therapeutic direction, and hereditary cancer risk assessment. This report presents a patient with an initial diagnosis of metastatic pancreatic adenocarcinoma (PDA), a disease with a dismal prognosis. Tumor sequencing revealed genomic abnormalities inconsistent with PDA, instead suggesting serous ovarian cancer. This molecular rediagnosis was further refined by the identification of a BRCA2 truncating mutation in the tumor, subsequently confirmed to be a germline event. These findings prompted the initiation of platinum-based chemotherapy, which produced a life-altering response, and referral to genetic counseling for her offspring. These results suggest that clinical tumor sequencing can simultaneously clarify diagnoses, guide therapy, and inform familial risk, even in patients with end-stage metastatic disease, making the case for the development of specific strategies to deploy sequencing coupled with big data in oncology to improve clinical cancer management.


Three-dimensional Force and Kinematic Interactions in V1 Skating at High Speeds.
To describe the detailed kinetics and kinematics associated with use of the V1 skating technique at high skiing speeds and to identify factors that predict performance. Fifteen elite male cross-country skiers performed an incremental roller-skiing speed test (Vpeak) on a treadmill using the V1 skating technique. Pole and plantar forces and whole-body kinematics were monitored at four submaximal speeds. The propulsive force of the "strong side" pole was greater than that of the "weak side" (P < 0.01), but no difference was observed for the legs. The poles generated approximately 44% of the total propulsion, being more effective than the legs in this respect (∼59% vs 11%, P < 0.001). Faster skiers exhibited more well-synchronized poling, exhibited more symmetric edging by and forces from the legs, and were more effective in transformation of resultant forces into propulsion. Cycle length was not correlated with either Vpeak or the impulse of total propulsive forces. The present findings provide novel insights into the coordination, kinetics, and kinematics of the arm and leg motion by elite athletes while V1 skating at high speeds. The faster skiers exhibit more symmetric leg motion on the "strong" and "weak" sides, as well as more synchronized poling. With respect to methods, the pressure insoles and three-dimensional kinematics in combination with the leg push-off model described here can easily be applied to all skating techniques, aiding in the evaluation of skiing techniques and comparison of effectiveness.


Models and impact of patient and public involvement in studies carried out by the Medical Research Council Clinical Trials Unit at University College London: findings from ten case studies.
Patient and public involvement (PPI) in studies carried out by the UK Medical Research Council Clinical Trials Unit (MRC CTU) at University College London varies by research type and setting. We developed a series of case studies of PPI to document and share good practice. We used purposive sampling to identify studies representing the scope of research at the MRC CTU and different approaches to PPI. We carried out semi-structured interviews with staff and patient representatives. Interview notes were analysed descriptively to categorise the main aims and motivations for involvement; activities undertaken; their impact on the studies and lessons learned. We conducted 19 interviews about ten case studies, comprising one systematic review, one observational study and 8 randomised controlled trials in HIV and cancer. Studies were either open or completed, with start dates between 2003 and 2011. Interviews took place between March and November 2014 and were updated in summer 2015 where there had been significant developments in the study (i.e. if the study had presented results subsequent to the interview taking place). A wide range of PPI models, including representation on trial committees or management groups, community engagement, one-off task-focused activities, patient research partners and participant involvement had been used. Overall, interviewees felt that PPI had a positive impact, leading to improvements, for example in the research question; study design; communication with potential participants; study recruitment; confidence to carry out or complete a study; interpretation and communication of results; and influence on future research. A range of models of PPI can benefit clinical studies. Researchers should consider different approaches to PPI, based on the desired impact and the people they want to involve. Use of multiple models may increase the potential impacts of PPI in clinical research.


Thyroid cancer in Friuli Venezia Giulia, northeastern Italy: incidence, overdiagnosis, and impact of type of surgery on survival.
Incidence rates of thyroid cancer (TC) increased in the last decades worldwide. This study aimed to describe TC incidence in the Friuli Venezia Giulia (FVG) region, to estimate the quota of overdiagnosis, and to investigate the impact of surgery on long-term survival after TC diagnosis. TC cases reported to the FVG population-based cancer registry during 2002-2013, aged <85 years, were included. Age standardized rates (ASR) on the European population were computed, while proportion of TC overdiagnosis was estimated in comparison with expected age-specific incidence rates from published time series. Adjusted hazard ratios of death, with 95% confidence intervals, were also estimated. During 2002-2013, 1701 TC cases were reported to the FVG cancer registry, with papillary TC (78.2%) as the most frequent histologic type. ASR increased from 12.4 to 16.5 in women and from 4.3 to 6.2 in men (+33.1% and +44.2%, respectively). Overdiagnosis was estimated as 79% of TC cases in women and 64% in men. Almost all TC cases (97.1%) underwent surgery, including 84.6% of women and 78.9% of men who underwent total thyroidectomy. Up to 10 years after TC diagnosis, the type of surgery did not appear to influence survival. This study documented an increase in TC incidence in FVG in the last decade, with overdiagnosis accounting for a large proportion of TC diagnoses and total thyroidectomy in more than 80% of cases. These findings suggest reconsidering thyroid screening practice and aggressive therapeutic strategies, as recommended by new TC guidelines.


Inspiratory resistive loading improves cycling capacity: a placebo controlled trial.
Respiratory muscle training has been shown to improve both its strength and endurance. The effect of these improvements on whole-body exercise performance remains controversial. To assess the effect of a 10 week inspiratory resistive loading (IRL) intervention on respiratory muscle performance and whole-body exercise endurance. Fifteen apparently healthy subjects (10 men, 5 women) were randomly allocated to one of three groups. One group underwent IRL set at 80% of maximum inspiratory pressure with ever decreasing work/rest ratios until task failure, for three days a week for 10 weeks (IRL group). A second placebo group performed the same training procedure but with a minimal resistance (PLA group). IRL and placebo training were performed at rest. The remaining five control subjects performed no IRL during the 10 week study period (CON group). Cycling endurance capacity at 75% VO(2)peak was measured before and after the intervention. After the 10 week IRL intervention, respiratory muscle strength (maximum inspiratory pressure) and endurance (sum of sustained maximum inspiratory pressure) had significantly improved (by 34% and 38% respectively). An increase in diaphragm thickness was also observed. These improvements translated into a 36% increase in cycling time to exhaustion at 75% VO(2)peak. During cycling trials, heart rate, ventilation, and rating of perceived exertion were attenuated in the IRL group. No changes were observed for the PLA or CON group either in the time to exhaustion or cardiorespiratory response to the same intensity of exercise. Ten weeks of IRL attenuated the heart rate, ventilatory, and perceptual response to constant workload exercise, and improved the cycling time to exhaustion. Familiarisation was not a factor and the placebo effect was minimal.


Changes in telomere length distribution in low-dose X-ray-irradiated human umbilical vein endothelial cells.
Ionizing radiation (IR) is known to be a cause of telomere dysfunction in tumor cells; however, very few studies have investigated X-ray-related changes in telomere length and the telomerase activity in normal human cells, such as umbilical vein endothelial cells (HUVECs). The loss of a few hundred base pairs from a shortened telomere has been shown to be important with respect to cellular senescence, although it may not be detected according to traditional mean telomere length [assessed as the terminal restriction fragment (TRF)] analyses. In the present study, a continuous time window from irradiation was selected to examine changes in the telomere length, including the mean TRF length, percentage of the telomere length, telomerase activity, apoptotic rate, and survival rate in HUVECs from the first day to the fourth day after the administration of a 0.5-Gy dose of irradiation. The mean TRF length in the irradiated HUVECs showed shorter telomere length in first 3 days, but they were not statistically significant. On the other hand, according to the percentage analysis of the telomere length, a decreasing tendency was noted in the longer telomere lengths (9.4-4.4 kb), with a significant increase in the shortest telomeres (4.4-2.3 kb) among the irradiated cells versus the controls from the first day to the third after irradiation; no significant differences were noted on the fourth day. These results suggest that the shortest telomeres are sensitive to the late stage of radiation damage. The proliferation of irradiated cells was suppressed after IR in contrast to the non-irradiated cells. The apoptotic rate was elevated initially both in IR- and non-IR-cells, but that of IR-cells was maintained at an elevated level thereafter in contrast to that of non-IR-cells decreasing promptly. Therefore, a 0.5-Gy dose of IR induces persistent apoptosis leading to an apparent growth arrest of the normal HUVECs.


Deletion analysis and clinical correlations in patients with Xp21 linked muscular dystrophy.
We carried out molecular deletion analysis on 142 patients with Duchenne/Becker muscular dystrophy which covered 25 exons of the dystrophin gene. We also evaluated the results by comparing with the clinical findings and examples in the literature. A deletion ratio of 63.7% was achieved. Exon 46 was the most frequently affected region. Interestingly we also observed four cases with muscle promoter (Mp) region deletions which have been rarely reported in the literature.


Water preparation practices in south Kalimantan, Indonesia.
This study investigated water preparation practices, water sources, and sanitation measures in 400 households in four villages in Banjar district of South Kalimantan, Indonesia. A survey of randomly selected households in each of the four villages was undertaken during the season of low prevalence of diarrhoeal disease (February-March 1989). A follow-up survey of the same households was conducted 6 months later to measure prevalence during the peak season. Twenty-three in-depth interviews were carried out with key persons in local villages. Observations on behaviour in the local food stalls were recorded. The results of the study indicate that 97% of the households report that they regularly boil their drinking water. However, 37% of the households regularly or occasionally mix boiled with unboiled water for drinking, or use unboiled water alone. The mixing of boiled with unboiled water is particularly frequent in the preparation of 'cold tea', a popular drink in households and food stalls. The occasional or regular use of unboiled water varies by ethnicity, education and literacy, and economic status. The use of unboiled water is associated with higher rates of childhood diarrhoeas in the households studied.


Turner syndrome and autoimmune diseases: record-linkage study.
There is increasing evidence that Turner syndrome is associated with an elevated risk of a range of autoimmune disorders. We aimed to document this in a national study. Use of a record-linked dataset of all hospital admissions in England, 1999-2011, to construct a retrospective cohort of people with Turner syndrome and a control cohort of people without it. Statistical follow-up to identify the occurrence of 29 separate autoimmune disorders in each cohort. Calculation of rate ratios, comparing the Turner and control cohorts. In the Turner syndrome cohort (2459 people), rate ratios were elevated for 16 of the 29 conditions. Examples included coeliac disease (rate ratio 14.0, 95% CI 10.2 to 18.8), Crohn's disease (5.3, 3.5 to 7.8), ulcerative colitis (3.9, 2.3 to 6.1), hypothyroidism (8.8, 7.8 to 9.9) and hyperthyroidism (4.9, 3.2 to 7.1). The increased risk of autoimmune disorders in people with Turner syndrome covers a wide range of conditions.


Outcome Measures for Acute/Subacute Cervical Sensorimotor Complete (AIS-A) Spinal Cord Injury During a Phase 2 Clinical Trial.
Effective treatment after cervical spinal cord injury (SCI) is imperative as so many activities of daily living (ADLs) are dependent on functional recovery of arm and hand actions. We focus on defining and comparing neurological and functional endpoints that might be used during acute or subacute Phase 2 clinical trials involving subjects with cervical sensorimotor complete SCI (ASIA Impairment Scale [AIS-A]). For the purposes of this review, the trial would examine the effects of a pharmaceutical small molecule, drug, biologic, or cell transplant on spinal tissue. Thus, neurological improvement is the intended consequence and is most directly measured by assessing neurological impairment (eg, motor aspects of the International Standards Neurological Classification of Spinal Cord Injury [ISNCSCI]). However, changes in neurological function, even if statistically significant, may not be associated with a clear functional impact (ie, a meaningful improvement in individual activity, such as independent self-care ADLs). The challenge is to measure improvement as precisely as possible (change in impairment), but to define a clinically meaningful response in the context of functional improvement (impact on activity limitations). The principal comparisons focused on elements of the ISNCSCI assessment, including upper extremity motor score and motor level. Personal activity capabilities were also examined at various time points. The data suggest that an improvement of 2 or more motor levels after cervical sensorimotor complete SCI may be a clinically meaningful endpoint threshold that could be used for acute and subacute Phase 2 trials with subjects having sensorimotor complete cervical SCI.


Suppressor of MEK null (SMEK)/protein phosphatase 4 catalytic subunit (PP4C) is a key regulator of hepatic gluconeogenesis.
Fasting promotes hepatic gluconeogenesis to maintain glucose homeostasis. The cAMP-response element binding protein (CREB)-regulated transcriptional coactivator 2 (CRTC2) is responsible for transcriptional activation of gluconeogenic genes and is critical for conveying the opposing hormonal signals of glucagon and insulin in the liver. Here, we show that suppressor of MEK null 1 (SMEK1) and SMEK2 [protein phosphatase 4 (PP4) regulatory subunits 3a and 3b, respectively] are directly involved in the regulation of hepatic glucose metabolism in mice. Expression of hepatic SMEK1/2 is up-regulated during fasting or in mouse models of insulin-resistant conditions in a Peroxisome Proliferator-Activated Receptor-gamma Coactivator 1α (PGC-1α)-dependent manner. Overexpression of SMEK promotes elevations in plasma glucose with increased hepatic gluconeogenic gene expression, whereas depletion of the SMEK proteins reduces hyperglycemia and enhances CRTC2 phosphorylation; the effect is blunted by S171A CRTC2, which is refractory to salt-inducible kinase (SIK)-dependent inhibition. Taken together, we would propose that mammalian SMEK/PP4C proteins are involved in the regulation of hepatic glucose metabolism through dephosphorylation of CRTC2.


Medical history, lifestyle, family history, and occupational risk factors for follicular lymphoma: the InterLymph Non-Hodgkin Lymphoma Subtypes Project.
Follicular lymphoma (FL) has been linked with cigarette smoking and, inconsistently, with other risk factors. We assessed associations of medical, hormonal, family history, lifestyle, and occupational factors with FL risk in 3530 cases and 22639 controls from 19 case-control studies in the InterLymph consortium. Age-, race/ethnicity-, sex- and study-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using logistic regression. Most risk factors that were evaluated showed no association, except for a few modest or sex-specific relationships. FL risk was increased in persons: with a first-degree relative with non-Hodgkin lymphoma (OR = 1.99; 95% CI = 1.55 to 2.54); with greater body mass index as a young adult (OR = 1.15; 95% CI = 1.04 to 1.27 per 5 kg/m(2) increase); who worked as spray painters (OR = 2.66; 95% CI = 1.36 to 5.24); and among women with Sjögren syndrome (OR = 3.37; 95% CI = 1.23 to 9.19). Lower FL risks were observed in persons: with asthma, hay fever, and food allergy (ORs = 0.79-0.85); blood transfusions (OR = 0.78; 95% CI = 0.68 to 0.89); high recreational sun exposure (OR = 0.74; 95% CI = 0.65 to 0.86, fourth vs first quartile); who worked as bakers or millers (OR = 0.51; 95% CI = 0.28 to 0.93) or university/higher education teachers (OR = 0.58; 95% CI = 0.41 to 0.83). Elevated risks specific to women included current and longer duration of cigarette use, whereas reduced risks included current alcohol use, hay fever, and food allergies. Other factors, including other autoimmune diseases, eczema, hepatitis C virus seropositivity, hormonal drugs, hair dye use, sun exposure, and farming, were not associated with FL risk. The few relationships observed provide clues suggesting a multifactorial etiology of FL but are limited in the extent to which they explain FL occurrence.


Effects of ozonated sewage effluent on reproduction and behavioral endpoints in zebrafish (Danio rerio).
Pharmaceutical residues and other micro-contaminants may enter aquatic environments through effluent from sewage treatment plants (STPs) and could cause adverse effects in wild fish. One strategy to alleviate this situation is to improve wastewater treatment by ozonation. To test the effectiveness of full-scale wastewater effluent ozonation at a Swedish municipal STP, the added removal efficiency was measured for 105 pharmaceuticals. In addition, gene expression, reproductive and behavioral endpoints were analyzed in zebrafish (Danio rerio) exposed on-site over 21 days to ozonated or non-ozonated effluents as well as to tap water. Ozone treatment (7 g O3/m3) removed pharmaceuticals by an average efficiency of 77% in addition to the conventional treatment, leaving 11 screened pharmaceuticals above detection limits. Differences in biological responses of the exposure treatments were recorded in gene expression, reproduction and behavior. Hepatic vitellogenin gene expression was higher in male zebrafish exposed to the ozonated effluent compared to the non-ozonated effluent and tap water treatments. The reproductive success was higher in fish exposed to ozonated effluent compared to non-ozonated effluent and to tap water. The behavioral measurements showed that fish exposed to the ozonated STP effluent were less active in swimming the first minute after placed in a novel vessel. Ozonation is a capable method for removing pharmaceuticals in effluents. However, its implementation should be thoroughly evaluated for any potential biological impact. Future research is needed for uncovering the factors which produced the in vivo responses in fish.


NR5A1 gene mutations: clinical, endocrine and genetic features in two girls with 46,XY disorder of sex development.
Steroidogenic factor 1, encoded by the NR5A1 gene, is a key regulator of endocrine function within the hypothalamic-pituitary-steroidogenic axis. Both homozygous, compound heterozygous and heterozygous mutations in the NR5A1 gene may determine 46,XY disorders of sex development (DSD). NR5A1 gene sequencing was performed in a cohort of 6 patients with 46,XY DSD without specific diagnosis. Heterozygous NR5A1 gene mutations were found in 2 girls, aged 0.5 years and 14 years. The older girl harbored the c.250C>T transition in exon 4 (p.Arg84Cys), previously reported in a Japanese girl. The younger girl presented a de novo novel exon 6 heterozygous frameshift mutation (c.1074dupG) in codon 359 associated with the p.Gly146Ala polymorphism the latter inherited from her father. This baby showed severe impairment of androgen secretion from the first months of life. Overt adrenal insufficiency did not occur, but the older girl showed subnormal cortisol peak after ACTH stimulation. NR5A1 gene mutations are a relatively frequent cause of 46,XY DSD in humans. Clear indications for management of these individuals remain elusive, mainly when diagnosis is made in infancy. Long-term monitoring of adrenal function should be recommended.


Severe embryotoxicity of artemisinin derivatives in experimental animals, but possibly safe in pregnant women.
Preclinical studies in rodents have demonstrated that artemisinins, especially injectable artesunate, can induce fetal death and congenital malformations at a low dose range. The embryotoxicity can be induced in those animals only within a narrow window in early embryogenesis. Evidence was presented that the mechanism by which embryotoxicity of artemisinins occurs seems to be limited to fetal erythropoiesis and vasculogenesis/ angiogenesis on the very earliest developing red blood cells, causing severe anemia in the embryos with higher drug peak concentrations. However, this embryotoxicity has not been convincingly observed in clinical trials from 1,837 pregnant women, including 176 patients in the first trimester exposed to an artemisinin agent or artemisinin-based combination therapy (ACT) from 1989 to 2009. In the rodent, the sensitive early red cells are produced synchronously over one day with single or multiple exposures to the drug can result in a high proportion of cell deaths. In contrast, primates required a longer period of treatment of 12 days to induce such embryonic loss. In humans only limited information is available about this stage of red cell development; however, it is known to take place over a longer time period, and it may well be that a limited period of treatment of 2 to 3 days for malaria would not produce serious toxic effects. In addition, current oral intake, the most commonly used route of administration in pregnant women with an ACT, results in lower peak concentration and shorter exposure time of artemisinins that demonstrated that such a concentration-course profile is unlikely to induce the embryotoxicity. When relating the animal and human toxicity of artemisinins, the different drug sensitive period and pharmacokinetic profiles as reviewed in the present report may provide a great margin of safety in the pregnant women.


Copper nitrate: a privileged reagent for organic synthesis.
Copper has been explored as an ideal candidate for replacing noble metals in organic synthesis, especially for practical large scale preparation. Recent decades have witnessed the renaissance and improvement of copper-catalyzed and copper-mediated organic reactions. Copper nitrate is a common inorganic copper salt which has been proved to be a ubiquitous reactant in organic synthesis due to its commercial availability, stability, inexpensiveness and environmentally benign nature. Copper nitrate could be used as a nitration reagent, oxidant, catalyst or promoter, and Lewis acid as well. Remarkably, great attention has been devoted to the efficient transformation of copper nitrate into functionalized or complicated compounds through various reaction types including cyclization, C-H activation, difunctionalization, nitration, rearrangement and asymmetric synthesis with chiral ligands. Further modification of copper nitrate, such as solid-supported copper nitrate or copper nitrate complexes, extends its applications in organic synthesis. The present review highlights recent advances of copper nitrate in organic synthesis, along with the mechanisms.


Testing a variable-length Cognitive Processing Therapy intervention for posttraumatic stress disorder in active duty military: Design and methodology of a clinical trial.
Combat-related trauma exposures have been associated with increased risk for posttraumatic stress disorder (PTSD) and comorbid mental health conditions. Cognitive Processing Therapy (CPT) is a 12-session manualized cognitive-behavioral therapy that has emerged as one of the leading evidence-based treatments for combat-related PTSD among military personnel and veterans. However, rates of remission have been less in both veterans and active duty military personnel compared to civilians, suggesting that studies are needed to identify strategies to improve upon outcomes in veterans of military combat. There is existing evidence that varying the number of sessions in the CPT protocol based on patient response to treatment improves outcomes in civilians. This paper describes the rationale, design, and methodology of a clinical trial examining a variable-length CPT intervention in a treatment-seeking active duty sample with PTSD to determine if some service members would benefit from a longer or shorter dose of treatment, and to identify predictors of length of treatment response to reach good end-state functioning. In addition to individual demographic and trauma-related variables, the trial is designed to evaluate factors related to internalizing/externalizing personality traits, neuropsychological measures of cognitive functioning, and biological markers as predictors of treatment response. This study attempts to develop a personalized approach to achieving positive treatment outcomes for service members suffering from PTSD. Determining predictors of treatment response can help to develop an adaptable treatment regimen that returns the greatest number of service members to full functioning in the shortest amount of time.


Priming with cold-adapted influenza A does not prevent infection but elicits long-lived protection against supralethal challenge with heterosubtypic virus.
We show in this study several novel features of T cell-based heterosubtypic immunity against the influenza A virus in mice. First, T cell-mediated heterosubtypic protection against lethal challenge can be generated by a very low priming dose. Second, it becomes effective within 5-6 days. Third, it provides protection against a very high dose challenge for >70 days. Also novel is the finding that strong, long-lasting, heterosubtypic protection can be elicited by priming with attenuated cold-adapted strains. We demonstrate that priming does not prevent infection of the lungs following challenge, but leads to earlier clearance of the virus and 100% survival after otherwise lethal challenge. Protection is dependent on CD8 T cells, and we show that CD4 and CD8 T cells reactive to conserved epitopes of the core proteins of the challenge virus are present after priming. Our results suggest that intranasal vaccination with cold-adapted, attenuated live virus has the potential to provide effective emergency protection against emerging influenza strains for several months.


The Multi-Fungicide Resistance Status of Aspergillus fumigatus Populations in Arable Soils and the Wider European Environment.
The evolution and spread of pan-azole resistance alleles in clinical and environmental isolates of Aspergillus fumigatus is a global human health concern. The identification of hotspots for azole resistance development in the wider environment can inform optimal measures to counteract further spread by minimizing exposure to azole fungicides and reducing inoculum build-up and pathogen dispersal. We investigated the fungicide sensitivity status of soil populations sampled from arable crops and the wider environment and compared these with urban airborne populations. Low levels of azole resistance were observed for isolates carrying the CYP51A variant F46Y/M172V/E427K, all belonging to a cluster of related cell surface protein (CSP) types which included t07, t08, t13, t15, t19, and t02B, a new allele. High levels of resistance were found in soil isolates carrying CYP51A variants TR34/L98H and TR46/Y121F/T289A, all belonging to CSP types t01, t02, t04B, or t11. TR46/Y121F/M172V/T289A/G448S (CSP t01) and TR46/Y121F/T289A/S363P/I364V/G448S (CSP t01), a new haplotype associated with high levels of resistance, were isolated from Dutch urban air samples, indicating azole resistance evolution is ongoing. Based on low numbers of pan-azole resistant isolates and lack of new genotypes in soils of fungicide-treated commercial and experimental wheat crops, we consider arable crop production as a coldspot for azole resistance development, in contrast to previously reported flower bulb waste heaps. This study also shows that, in addition to azole resistance, several lineages of A. fumigatus carrying TR-based CYP51A variants have also developed acquired resistance to methyl benzimidazole carbamate, quinone outside inhibitor and succinate dehydrogenase (Sdh) inhibitor fungicides through target-site alterations in the corresponding fungicide target proteins; beta-tubulin (F200Y), cytochrome b (G143A), and Sdh subunit B (H270Y and H270R), respectively. Molecular typing showed that several multi-fungicide resistant strains found in agricultural soils in this study were clonal as identical isolates have been found earlier in the environment and/or in patients. Further research on the spread of different fungicide-resistant alleles from the wider environment to patients and vice versa can inform optimal practices to tackle the further spread of antifungal resistance in A. fumigatus populations and to safeguard the efficacy of azoles for future treatment of invasive aspergillosis.


Relations of arterial stiffness and endothelial function to brain aging in the community.
To determine the association of arterial stiffness and pressure pulsatility, which can damage small vessels in the brain, with vascular and Alzheimer-type brain aging. Stroke- and dementia-free Framingham Offspring Study participants (n = 1,587, 61 ± 9 years, 45% male) underwent study of tonometric arterial stiffness and endothelial function (1998-2001) and brain MRI and cognition (1999-2002). We related carotid-femoral pulse wave velocity (CFPWV), mean arterial and central pulse pressure, and endothelial function to vascular brain aging by MRI (total cerebral brain volume [TCBV], white matter hyperintensity volume, silent cerebral infarcts) and vascular and Alzheimer-type cognitive aging (Trails B minus Trails A and logical memory-delayed recall, respectively). Higher CFPWV was associated with lower TCBV, greater white matter hyperintensity volume, and greater prevalence of silent cerebral infarcts (all p < 0.05). Each SD greater CFPWV was associated with lower TCBV equivalent to 1.2 years of brain aging. Mean arterial and central pulse pressure were associated with greater white matter hyperintensity volume (p = 0.005) and lower TCBV (p = 0.02), respectively, and worse verbal memory (both p < 0.05). Associations of tonometry variables with TCBV and white matter hyperintensity volume were stronger among those aged 65 years and older vs those younger than 65 years (p < 0.10 for interaction). Brachial artery endothelial function was unrelated to MRI measures (all p > 0.05). Greater arterial stiffness and pressure pulsatility are associated with brain aging, MRI vascular insults, and memory deficits typically seen in Alzheimer dementia. Future investigations are warranted to evaluate the potential impact of prevention and treatment of unfavorable arterial hemodynamics on neurocognitive outcomes.


Novel preparation techniques for controlling microbubble uniformity: a comparison.
In recent years, there has been increasing interest in the use of coated microbubbles as vehicles for ultrasound mediated targeted drug delivery. This application requires a high degree of control over the size and uniformity of microbubbles, in order to ensure accurate dosing of a given drug and to maximise delivery efficiency. Similarly, as more advanced imaging techniques are developed which exploit the complex nonlinear features of the microbubble signal and/or enable quantification of tissue perfusion, the ability to predetermine the acoustic response of a microbubble suspension is becoming increasingly important. Consequently, a number of new preparation technologies have been developed to meet the demand for improved control over microbubble characteristics. The aim of the work described in this paper was to compare a conventional microbubble preparation technique, sonication, with two more recent methods: coaxial electrohydrodynamic atomisation and microfluidic (T-junction) processing, in terms of their ability to produce bubbles which are sufficiently small and stable for in vivo use, microbubble uniformity, relative production rates and other practical and economic considerations.


Understanding the uptake of cervical cancer screening: the contribution of the health belief model.
The health belief model, explaining health and illness behaviour, is 25 years old. Criticisms of the model have included its abstract nature and its emphasis on the rationality of patients' behaviour. Its lack of predictive power means it provides a useful framework rather than a true model. The health belief model is used here to review the literature that has advanced our understanding of the factors affecting uptake of cervical screening. The influence of age and social class on perceptions of vulnerability, and the costs and benefits of screening are highlighted. The body of work reviewed has helped expose inherent limitations of screening programmes. The main obstacles to the success of cervical screening are organizational, for example, the inaccuracy of address registers. Numerous ways of encouraging uptake are identified. These include appropriately worded invitations and educational material, personalized approaches from members of the primary health care team and flexible surgery hours. The incentives introduced under the 1990 general practitioner contract are likely to help increase uptake.


Biomechanical consequences of total plantar fasciotomy: a review of the literature.
Plantar fascia release for chronic plantar fasciitis has provided excellent pain relief and rapid return to activities with few reported complications. Cadaveric studies have led to the identification of some potential postoperative problems, most commonly weakness of the medial longitudinal arch and pain in the lateral midfoot. An electronic search was conducted of the MEDLINE, ScienceDirect, SportDiscus, EMBASE, CINAHL, Cochrane, and AMED databases. The keywords used to search these databases were plantar fasciotomy and medial longitudinal arch. Articles published between 1976 and 2008 were identified. Collectively, results of cadaveric studies suggested that plantar fasciotomy leads to loss of integrity of the medial longitudinal arch and that total plantar fasciotomy is more detrimental to foot structure than is partial fasciotomy. In vivo studies, although limited in number, concluded that although clinical outcomes were satisfactory, medial longitudinal arch height decreased and the center of pressure of the weightbearing foot was excessively medially deviated postoperatively. Plantar fasciotomy, in particular total plantar fasciotomy, may lead to loss of stability of the medial longitudinal arch and abnormalities in gait, in particular an excessively pronated foot. Further in vivo studies on the long-term biomechanical effects of plantar fasciotomy are required.


A peptide-based target screen implicates the protein kinase CK2 in the global regulation of caspase signaling.
The convergence of caspase and protein kinase signaling pathways has become increasingly evident, as illustrated by the protection of caspase substrates from cleavage upon undergoing phosphorylation at or near to their caspase recognition motifs. To investigate the global role of phosphorylation in the regulation of caspase signaling, we designed a peptide match program to identify sequences from the human proteome that contained overlapping recognition motifs for caspases and kinases. We identified the protein kinase CK2 as the most prominent kinase with a consensus site for phosphorylation that overlapped with caspase recognition motifs. We then evaluated potential targets of CK2 and caspases by combining peptide array target screens with identification of caspase substrates. We identified numerous shared candidate targets of CK2 and caspases, including procaspase-3, which functions at a level at which both intrinsic and extrinsic apoptotic signals converge. Together, these data support a role for CK2-dependent phosphorylation as a global mechanism for inhibiting caspase signaling pathways.


The clinical effectiveness of concise cognitive behavioral therapy with or without pharmacotherapy for depressive and anxiety disorders; a pragmatic randomized controlled equivalence trial in clinical practice.
Depressive and anxiety disorders contribute to a high disease burden. This paper investigates whether concise formats of cognitive behavioral- and/or pharmacotherapy are equivalent with longer standard care in the treatment of depressive and/or anxiety disorders in secondary mental health care. A pragmatic randomized controlled equivalence trial was conducted at five Dutch outpatient Mental Healthcare Centers (MHCs) of the Regional Mental Health Provider (RMHP) 'Rivierduinen'. Patients (aged 18-65 years) with a mild to moderate anxiety and/or depressive disorder, were randomly allocated to concise or standard care. Data were collected at baseline, 3, 6 and 12 months by Routine Outcome Monitoring (ROM). Primary outcomes were the Brief Symptom Inventory (BSI) and the Web Screening Questionnaire (WSQ). We used Generalized Estimating Equations (GEE) to assess outcomes. Between March 2010 and December 2012, 182 patients, were enrolled (n=89 standard care; n=93 concise care). Both intention-to-treat and per-protocol analyses demonstrated equivalence of concise care and standard care at all time points. Severity of illness reduced, and both treatments improved patient's general health status and subdomains of quality of life. Moreover, in concise care, the beneficial effects started earlier. Concise care has the potential to be a feasible and promising alternative to longer standard secondary mental health care in the treatment of outpatients with a mild to moderate depressive and/or anxiety disorder. For future research, we recommend adhering more strictly to the concise treatment protocols to further explore the beneficial effects of the concise treatment. The study is registered in the Netherlands Trial Register, number NTR2590. Clinicaltrials.gov identifier: NCT01643642.


Chiral 2D Organic Inorganic Hybrid Perovskite with Circular Dichroism Tunable Over Wide Wavelength Range.
The effect of chemical-composition modification on the chiroptical property of chiral organic ammonium cation-containing organic inorganic hybrid perovskite (chiral OIHP) is investigated. Varying the mixing ratio of bromide and iodide anions in S- or R-C6H5CH2(CH3)NH3)2PbI4(1-x)Br4x modifies the band gap of chiral OIHP, leading to a shift of the circular dichroism (CD) signal from 495 to 474 nm. However, it is also found that an abrupt crystalline structure transition occurs, and the CD signal is turned off when iodide-determinant phases are transformed into the bromide-determinant phase. To obtain CD in the wavelength range where the bromide-determinant phase is supposed to exhibit chiroptical activity, that is, <474 nm, S- or R-C12H7CH2(CH3)NH3 with a larger spacer group can be adopted; thus, the CD signal can be further blue-shifted to ∼375 nm. Here, we show that chemical-composition modification of chiral OIHP affects the chiroptical properties of chiral OIHP in two ways: (1) tuning the wavelength of CD by modulating the excitonic band structure and (2) switching the CD on and off by inducing a crystalline-structure change. These properties can be utilized for structural engineering of high-performance chiroptical materials for spin-polarized light-emitting devices and polarization-based optoelectronics.


Molecular clustering identifies complement and endothelin induction as early events in a mouse model of glaucoma.
Glaucoma is one of the most common neurodegenerative diseases. Despite this, the earliest stages of this complex disease are still unclear. This study was specifically designed to identify early stages of glaucoma in DBA/2J mice. To do this, we used genome-wide expression profiling of optic nerve head and retina and a series of computational methods. Eyes with no detectable glaucoma by conventional assays were grouped into molecularly defined stages of disease using unbiased hierarchical clustering. These stages represent a temporally ordered sequence of glaucoma states. We then determined networks and biological processes that were altered at these early stages. Early-stage expression changes included upregulation of both the complement cascade and the endothelin system, and so we tested the therapeutic value of separately inhibiting them. Mice with a mutation in complement component 1a (C1qa) were protected from glaucoma. Similarly, inhibition of the endothelin system with bosentan, an endothelin receptor antagonist, was strongly protective against glaucomatous damage. Since endothelin 2 is potently vasoconstrictive and was produced by microglia/macrophages, our data provide what we believe to be a novel link between these cell types and vascular dysfunction in glaucoma. Targeting early molecular events, such as complement and endothelin induction, may provide effective new treatments for human glaucoma.


Finding Bugs in Cryptographic Hash Function Implementations.
Cryptographic hash functions are security-critical algorithms with many practical applications, notably in digital signatures. Developing an approach to test them can be particularly difficult, and bugs can remain unnoticed for many years. We revisit the NIST hash function competition, which was used to develop the SHA-3 standard, and apply a new testing strategy to all available reference implementations. Motivated by the cryptographic properties that a hash function should satisfy, we develop four tests. The Bit-Contribution Test checks if changes in the message affect the hash value, and the Bit-Exclusion Test checks that changes beyond the last message bit leave the hash value unchanged. We develop the Update Test to verify that messages are processed correctly in chunks, and then use combinatorial testing methods to reduce the test set size by several orders of magnitude while retaining the same fault-detection capability. Our tests detect bugs in 41 of the 86 reference implementations submitted to the SHA-3 competition, including the rediscovery of a bug in all submitted implementations of the SHA-3 finalist BLAKE. This bug remained undiscovered for seven years, and is particularly serious because it provides a simple strategy to modify the message without changing the hash value returned by the implementation. We detect these bugs using a fully-automated testing approach.


Differentially methylated regions of imprinted genes in prenatal, perinatal and postnatal human tissues.
Epigenetic plasticity in relation to in utero exposures may mechanistically explain observed differences in the likelihood of developing common complex diseases including hypertension, diabetes and cardiovascular disease through the cumulative effects of subtle alterations in gene expression. Imprinted genes are essential mediators of growth and development and are characterized by differentially methylated regulatory regions (DMRs) that carry parental allele-specific methylation profiles. This theoretical 50% level of methylation provides a baseline from which endogenously- or exogenously-induced deviations in methylation can be detected. We quantified DNA methylation at imprinted gene DMRs in a large panel of human conceptal tissues, in matched buccal cell specimens collected at birth and at one year of age, and in the major cell fractions of umbilical cord blood to assess the stability of methylation at these regions. DNA methylation was measured using validated pyrosequencing assays at seven DMRs regulating the IGF2/H19, DLK1/MEG3, MEST, NNAT and SGCE/PEG10 imprinted domains. DMR methylation did not significantly differ for the H19, MEST and SGCE/PEG10 DMRs across all conceptal tissues analyzed (ANOVA p>0.10). Methylation differences at several DMRs were observed in tissues from brain (IGF2 and MEG3-IG DMRs), liver (IGF2 and MEG3 DMRs) and placenta (both DLK1/MEG3 DMRs and NNAT DMR). In most infants, methylation profiles in buccal cells at birth and at one year of age were comparable, as was methylation in the major cell fractions of umbilical cord blood. Several infants showed temporal deviations in methylation at multiple DMRs. Similarity of inter-individual and intra-individual methylation at some, but not all of the DMRs analyzed supports the possibility that methylation of these regions can serve as useful biosensors of exposure.


A shorter telomere is the key factor in preventing cultured human mesenchymal stem cells from senescence escape.
Mesenchymal stem cells (MSCs) from various animals undergo spontaneous transformation in vitro,establishing some malignant characteristics. However,this phenomenon seems seldom appearing in human (h)MSCs. To address the question whether the hMSCs really do not undergo the spontaneous transformation and why,the present study compared MSCs from two species under the same conditions, the commercialized primary hMSCs whose in vitro life span is very uniform, and the rat (r)MSCs whose spontaneous transformation in vitro is well defined.It was demonstrated that in rMSCs, there were small numbers of re-proliferating cells appearing after a substantial senescent period. These "senescence-escaped"rMSCs were highly proliferative and did not show any sign of growth arrest during the following subcultures upto observed passage 32. Whereas after entering senescence, hMSCs no longer re-proliferated and finally died from apoptosis. Compared with rMSCs, the hMSCs possessed a much shorter telomere, and lacked both telomerase reverse transcriptase expression and telomerase activity. When proliferating from pre-senescent to senescent stages,the hMSCs had a greater loss of relative telomere length(51 % in hMSC vs. 15 % in rMSC), but both cells displayed a similar average telomere shortening per population doubling (0.50 ± 0.06 kb in rMSC vs. 0.49 ± 0.06 kbin hMSC; p > 0.05), indicating that the greater relative shortening of the hMSC telomeres was due to their original shorter length, rather than lack of telomere maintenance mechanisms. In conclusion, the hMSCs do not spontaneously initiate transformation, because they cannot escape senescence. This is particularly due to their much shorter telomere.


Dopamine Transporter Dynamics of N-Substituted Benztropine Analogs with Atypical Behavioral Effects.
Atypical dopamine transporter (DAT) inhibitors, despite high DAT affinity, do not produce the psychomotor stimulant and abuse profile of standard DAT inhibitors such as cocaine. Proposed contributing features for those differences include off-target actions, slow onsets of action, and ligand bias regarding DAT conformation. Several 3α-(4',4''-difluoro-diphenylmethoxy)tropanes were examined, including those with the following substitutions: N-(indole-3''-ethyl)- (GA1-69), N-(R)-2''-amino-3''-methyl-n-butyl- (GA2-50), N-2''aminoethyl- (GA2-99), and N-(cyclopropylmethyl)- (JHW013). These compounds were previously reported to have rapid onset of behavioral effects and were presently evaluated pharmacologically alone or in combination with cocaine. DAT conformational mode was assessed by substituted-cysteine accessibility and molecular dynamics (MD) simulations. As determined by substituted-cysteine alkylation, all BZT analogs except GA2-99 showed bias for a cytoplasmic-facing DAT conformation, whereas cocaine stabilized the extracellular-facing conformation. MD simulations suggested that several analog-DAT complexes formed stable R85-D476 "outer gate" bonds that close the DAT to extracellular space. GA2-99 diverged from this pattern, yet had effects similar to those of other atypical DAT inhibitors. Apparent DAT association rates of the BZT analogs in vivo were slower than that for cocaine. None of the compounds was self-administered or stimulated locomotion, and each blocked those effects of cocaine. The present findings provide more detail on ligand-induced DAT conformations and indicate that aspects of DAT conformation other than "open" versus "closed" may facilitate predictions of the actions of DAT inhibitors and may promote rational design of potential treatments for psychomotor-stimulant abuse.


Malignant pleural effusion management: keeping the flood gates shut.
With no cure for malignant pleural effusion, efforts are focused on symptomatic management. Historically, this symptomatic management was achieved with the instillation of a sclerosant agent into the pleural space to achieve pleurodesis. The development of the tunnelled indwelling pleural catheter and ambulatory pleural drainage changed the management of malignant pleural effusion, not solely by offering an alternative management pathway, but by challenging how health-care providers view success in a palliative condition. Furthermore, with additional treatment options available, increased imperative exists to better characterise patients to enable a personalised approach to their care. We have done a review of the scientific literature and clinical trial registries to provide an overview of the current and ground-breaking research published in the past 10 years.


Rett syndrome like phenotypes in the R255X Mecp2 mutant mouse are rescued by MECP2 transgene.
Rett syndrome (RTT) is a severe neurodevelopmental disorder that is usually caused by mutations in Methyl-CpG-binding Protein 2 (MECP2). Four of the eight common disease causing mutations in MECP2 are nonsense mutations and are responsible for over 35% of all cases of RTT. A strategy to overcome disease-causing nonsense mutations is treatment with nonsense mutation suppressing drugs that allow expression of full-length proteins from mutated genes with premature in-frame stop codons. To determine if this strategy is useful in RTT, we characterized a new mouse model containing a knock-in nonsense mutation (p.R255X) in the Mecp2 locus (Mecp2(R255X)). To determine whether the truncated gene product acts as a dominant negative allele and if RTT-like phenotypes could be rescued by expression of wild-type protein, we genetically introduced an extra copy of MECP2 via an MECP2 transgene. The addition of MECP2 transgene to Mecp2(R255X) mice abolished the phenotypic abnormalities and resulted in near complete rescue. Expression of MECP2 transgene Mecp2(R255X) allele also rescued mTORC1 signaling abnormalities discovered in mice with loss of function and overexpression of Mecp2. Finally, we treated Mecp2(R255X) embryonic fibroblasts with the nonsense mutation suppressing drug gentamicin and we were able to induce expression of full-length MeCP2 from the mutant p.R255X allele. These data provide proof of concept that the p.R255X mutation of MECP2 is amenable to the nonsense suppression therapeutic strategy and provide guidelines for the extent of rescue that can be expected by re-expressing MeCP2 protein.


Performance of deep neural network-based artificial intelligence method in diabetic retinopathy screening: a systematic review and meta-analysis of diagnostic test accuracy.
Automatic diabetic retinopathy screening system based on neural networks has been used to detect diabetic retinopathy (DR). However, there is no quantitative synthesis of performance of these methods. We aimed to estimate the sensitivity and specificity of neural networks in DR grading. Medline, Embase, IEEE Xplore, and Cochrane Library were searched up to 23 July 2019. Studies that evaluated performance of neural networks in detection of moderate or worse DR or diabetic macular edema using retinal fundus images with ophthalmologists' judgment as reference standard were included. Two reviewers extracted data independently. Risk of bias of eligible studies was assessed using QUDAS-2 tool. Twenty-four studies involving 235 235 subjects were included. Quantitative random-effects meta-analysis using the Rutter and Gatsonis hierarchical summary receiver operating characteristics (HSROC) model revealed a pooled sensitivity of 91.9% (95% CI: 89.6% to 94.3%) and specificity of 91.3% (95% CI: 89.0% to 93.5%). Subgroup analyses and meta-regression did not provide any statistically significant findings for the heterogeneous diagnostic accuracy in studies with different image resolutions, sample sizes of training sets, architecture of convolutional neural networks, or diagnostic criteria. State-of-the-art neural networks could effectively detect clinical significant DR. To further improve diagnostic accuracy of neural networks, researchers might need to develop new algorithms rather than simply enlarge sample sizes of training sets or optimize image quality.


Patch test results of hand eczema patients: relation to clinical types.
Allergic contact dermatitis is a well-known cause of hand eczema, although the influence of contact allergens on different clinical types of hand eczema remains still unclear. To identify most common positive tested allergens among hand eczema patients and to define the relation between specific contact allergies and clinical types of hand eczema according to the guidelines of the Danish Contact Dermatitis Group (DCDG). We included 1571 hand eczema subjects who were patch tested from 1 January 2002 to 31 December 2013. They were retrospectively classified according to the guidelines of the DCDG into six clinical types: recurrent vesicular hand eczema, chronic fissured hand eczema, hyperkeratotic palmar eczema, pulpitis, interdigital eczema and nummular hand eczema according to a newly developed flow chart. The prevalence of sensitizations and association with clinical type, atopic dermatitis, age and gender were studied. A total of 1395 subjects were classified into one of the six clinical types. The most frequently found clinical types were recurrent vesicular hand eczema (39.7%) and chronic fissured hand eczema (35.5%). Subjects with recurrent vesicular hand eczema were significantly more likely to have a contact allergy (OR 1.55), whereas subjects with hyperkeratotic palmar eczema and pulpitis were less likely to be sensitized (OR 0.51; OR 0.44). Overall, metals (nickel sulphate, cobalt chloride), fragrances and preservatives (methylchloroisothiazoline/methylisothiazoline, methyldibromoglutaronitrile) were the most frequent sensitizers in patients with hand eczema. This did not deviate in the different clinical types, although subjects with recurrent vesicular hand eczema were significantly more frequently sensitized to nickel sulphate and other allergens compared to other clinical types of hand eczema. In the diagnostic work up of hand eczema subjects with recurrent vesicular hand eczema should be patch tested, especially women of older age, although the need for patch testing in males with hyperkeratotic palmar eczema might be less imperative.


MRI abnormalities of the hippocampus and cavum septi pellucidi in females with schizotypal personality disorder.
This study examined MRI hippocampal volume and cavum septi pellucidi (CSP) in female subjects with schizotypal personality disorder (SPD) and comparison subjects. MRI was performed on 20 SPD and 29 comparison subjects with delineation of left and right hippocampi. Number of slices containing the CSP was counted. Subjects were given a working memory task, the Delayed Alternation task and other measures of working memory including the Wechsler Memory Test-Revised and the California Verbal Learning Test. Clinical measures were derived from the SCID-II. SPD females evinced bilaterally smaller hippocampal volumes compared with non-psychiatric female subjects (15.1% on left, 15.7% on right). Additionally, SPD subjects showed statistically significantly more slices containing CSP, and a trend level difference when large CSP was defined as four or more slices (20% vs. 6.9%). SPD subjects demonstrated more errors, more perseverations, and a trend toward more failure to maintain set on the Delayed Alternating task, which were associated with smaller left hippocampal volumes. There was no difference between groups in logical memory, verbal learning or semantic clustering nor a significant correlation between these measures and hippocampal volumes. Clinically, in SPD subjects, right hippocampal volumes correlated negatively with odd appearance/behavior and positively with suspiciousness/paranoia, and odd speech was positively correlated with the number of slices containing a CSP in exploratory analyses. Female SPD subjects showed bilaterally smaller hippocampal volumes and larger CSP than comparison subjects, similar to what has been shown in schizophrenia. Moreover, these abnormalities have clinically significant associations which may help to explain some of the manifestations of the disorder.


Presence of T 145 on cytolytic T cell lines and their lectin-resistant mutants.
Murine cytolytic T cell lines have been analyzed for the expression of two surface glycoproteins called T145 and T130. T145, known to be expressed by activated cytolytic T cells, is also expressed by such lines, but T130, which has been described by a universal T cell marker, is not. Our results suggest a structural relationship between T145 and T130. Vicia villosa lectin, which binds selectively to T145 of activated T cells and which is cytotoxic for cytolytic T cell lines, has been used to select lectin-resistant mutants from these lines. Five independent lectin-resistant mutants have been obtained. All of them are cytolytically active, bind up to 100-fold less lectin than the parental lines, but still express T145 or a closely related glycoprotein.


Representation of Naturalistic Image Structure in the Primate Visual Cortex.
The perception of complex visual patterns emerges from neuronal activity in a cascade of areas in the primate cerebral cortex. We have probed the early stages of this cascade with "naturalistic" texture stimuli designed to capture key statistical features of natural images. Humans can recognize and classify these synthetic images and are insensitive to distortions that do not alter the local values of these statistics. The responses of neurons in the primary visual cortex, V1, are relatively insensitive to the statistical information in these textures. However, in the area immediately downstream, V2, cells respond more vigorously to these stimuli than to matched control stimuli. Humans show blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI responses in V1 and V2) that are consistent with the neuronal measurements in macaque. These fMRI measurements, as well as neurophysiological work by others, show that true natural scenes become a more prominent driving feature of cortex downstream from V2. These results suggest a framework for thinking about how information about elementary visual features is transformed into the specific representations of scenes and objects found in areas higher in the visual pathway.


Relationship of neuroimaging to typical sleep times during a clinical reasoning task: a pilot study.
Sleep deprivation and fatigue have been associated with medical errors, clinical performance decrements, and reduced quality of life for both practicing physicians and medical students. Greater understanding of the impact of sleep quantity on clinical reasoning could improve patient care. The purpose of our pilot study was to examine relationships between clinical reasoning (assessed by functional magnetic resonance imaging) and sleep time (measured in different ways by actigraphy) while answering multiple-choice questions (MCQs) from licensing agencies. Residents and faculty were administered a clinical reasoning exercise (MCQs from licensing bodies) during functional magnetic resonance imaging. Usual sleep patterns were sampled with actigraphy. Covariate analysis was used to examine the relationship between sleep duration (mean sleep, minimum sleep, maximum sleep) and brain activity during clinical reasoning (solving MCQs from licensing bodies). The mean sleep time over the duration of monitoring for the group was 7.19 hours (SD 0.66) with a range of 6.1 to 8.1 hours (internal medicine faculty 7.1 hours, SD 0.41; internal medicine residents 7.27 hours, SD 0.92). There was a negative relationship between activation in the prefrontal cortex and minimum sleep time while reflecting on MCQs. Our findings provide evidence that the quantity of sleep can modulate brain activity while completing a clinically meaningful task that should be confirmed in larger studies. Our findings suggest that the construct of sleepiness may be more complex than appreciated by many and that the most important of these sleep measures in terms of outcomes remains to be determined.


Chronic obstructive pulmonary disease (COPD) and lung cancer screening.
The primary aim of lung cancer screening is to improve survival from lung cancer by identifying early stage non-small cell lung cancers and prolong survival through their surgical removal. In a post-hoc analysis of 10,054 screening participants from the National Lung Screening Trial (NLST) we show that the risk of lung cancer, according to the PLCOm2012 model, is closely related to the likelihood of having chronic obstructive pulmonary disease (COPD). Those at greatest risk for lung cancer have the highest prevalence of COPD and greater likelihood of dying of a non-lung cancer cause. This "competing cause of death" effect occurs because smokers eligible for lung cancer screening have a high prevalence of comorbid disease and greater likelihood of dying from cardiovascular disease, respiratory disease or other cancers. This means high risk smokers at greatest risk of lung cancer may not necessarily benefit from screening due to greater inoperability and premature death. In this analysis we show that the benefit of annual computed tomography (CT) screening is greatest in those with normal lung function or only mild-to-moderate COPD. We found no mortality benefit in those with severe or very severe COPD (GOLD 3-4). We also show that the efficiency of screening, based on optimizing the number of lung cancer deaths averted per 1,000 persons screened, is best achieved by screening those at intermediate risk. By combining clinical risk variables with a gene-based risk score, even greater reductions in lung cancer mortality can be achieved with CT. We suggest a biomarker-led outcomes-based approach may help to better define which eligible smokers might defer screening (low risk of lung cancer), discontinue screening (high risk of overtreatment with little benefit) or continue screening to achieve the greatest reduction in lung cancer mortality.


Imaging of intracellular sodium with shift reagent aided (23)Na CSI in isolated rat hearts.
23Na chemical shift imaging (CSI) in conjunction with shift reagents was used to obtain images of intracellular (Na(i)) and extracellular sodium (Na(e)) in isolated rat hearts. It was demonstrated that the increase of Na(i) concentration in ischemic myocardium can be detected with this technique. 3D acquisition-weighted (23)Na CSI datasets with a nominal spatial resolution of 1.7 x 1.7 x 2.9 mm were acquired in 30 min in normoxic hearts and in globally or locally ischemic hearts. The shift reagent Tm(DOTP)(5-) was used to discriminate Na(i) and Na(e) signals. Na(i) maps could be generated in ischemic hearts, but not in normoxic hearts as the signal-to-noise ratio is too low. The Na(i) signal increased by more than 100% and the Na(e) signal decreased by more than 50% in myocardium of globally ischemic hearts (n = 3) compared to normoxic hearts (n = 3). In hearts with an acute occlusion of the left anterior descending coronary artery (n = 3), there was a local Na(i) signal increase in the anterior wall in the range of 60-110% compared to remote, normoxic tissue.