Observational study of dermatological manifestations in patients admitted to a tertiary poison center in Iran
Background/aim: Acute unintentional and deliberate poisoning by medications and chemicals is a frequent emergency, especially in Iran. This study aimed to evaluate the frequency and character of skin findings occurring in patients with acute intentional and aunintentional poisoning. Materials and methods: This prospective observational study was performed at the Loghman Hakim Hospital Poison Center over a period of 6 months from April 2016 to September 2016. Data including patient demographics, cause of poisoning, and level of consciousness were collected. Pediatric patients (under the age of 13) and patients who died in the first hours of admission were excluded from the study. Results: The most common cause of toxicity-related admission in our patients was methadone overdose. The most common skin finding in these patients was xerosis. According to our results, there was an association between tramadol poisoning and self-induced lesions. Shin hyperpigmentation was found to be significantly more frequent in patients with lead poisoning. Conclusion: Further study is recommended to shed light on the possible association of drug poisoning and skin lesions.


Plasmon-assisted grafting of anisotropic nanoparticles - spatially selective surface modification and the creation of amphiphilic SERS nanoprobes.
Amphiphilic nanoparticles (NPs) with a spatially selective distribution of grafted functional groups have great potential in the field of sensing, advanced imaging, and therapy due to their unique surface properties. The main techniques for the spatially selective functionalization of NPs utilize the surface-assisted approaches, which significantly restrict their production throughput. In this work, we propose an alternative plasmon-based route for the spatially selective grafting of anisotropic gold nanorods (AuNRs) using iodonium and diazonium salts. Utilization of longer laser wavelengths leads to the excitation of longitudinal plasmon resonances on AuNR tips, plasmon-assisted homolysis of the C-I bond in iodonium salts and the formation of aryl radicals, which are further grafted to the tips of AuNRs. The sides of AuNRs were subsequently decorated through spontaneous diazonium surface grafting. As a result, the AuNRs with spatially separated functional groups were prepared in a versatile way, primarily in solution and without the need for a sophisticated technique of NP immobilization or surface screening. The versatility of the proposed approach was proved on three kinds of AuNRs with different architectures and wavelength positions of plasmon absorption bands. Moreover, the applicability of the prepared amphiphilic AuNRs was shown by efficient trapping and SERS sensing of amphiphilic biomolecules.


Primary hormonogenic sites as conserved autoepitopes on thyroglobulin in murine autoimmune thyroiditis. Secondary role of iodination.
We hypothesized earlier that conserved T cell epitopes and those unique to mouse thyroglobulin (MTg) contributed to its total thyroiditogenicity in murine autoimmune thyroiditis. Recent studies of synthetic peptides from human Tg (HTg) revealed no immunodominant epitopes. The role of iodine residues, considered by some to render Tg immunogenic, became unclear, since only one 12-mer peptide contained thyroxine (T4) positioned at hormonogenic site 2553. It primed T cells for thyroiditis transfer, but noniodinated peptide containing thyronine (T0) was not compared. To determine 1) whether other primary hormonogenic sites were likewise immunogenic and 2) whether iodination was requisite for this and other sites to be an autoepitope, we derivatized three pairs of 12-mer peptides, 1-12, 2549-2560, 2559-2570, containing T0 or T4 at positions 5, 2553, or 2567, respectively. The six peptides were used to stimulate MTg-primed cells in vitro and to immunize mice. None directly induced thyroiditis; peptide Abs were the lowest in mice given hT0(2567) or hT4(2567). Of the three T4-containing peptides, hT4(5) and hT4(2553), but not hT4(2567), stimulated MTg-primed or HTg-primed T cells in vitro, with hT4(2553) being the stronger. Comparing hT0(2553) with hT4(2553), both activated MTg-primed, or peptide-primed, T cells to transfer thyroiditis. The marked immunogenicity of noniodinated hT0(2553) and the poor antigenicity of hT4(5) and hT4(2567) demonstrate that immunogenicity of a conserved hormonogenic site is dependent more on its amino acid sequence than on T4 substitution. Iodination may enhance antigenicity and/or binding affinity, but it is not required for a Tg hormonogenic site to be an autoepitope.


Multilevel summation with B-spline interpolation for pairwise interactions in molecular dynamics simulations.
The multilevel summation method for calculating electrostatic interactions in molecular dynamics simulations constructs an approximation to a pairwise interaction kernel and its gradient, which can be evaluated at a cost that scales linearly with the number of atoms. The method smoothly splits the kernel into a sum of partial kernels of increasing range and decreasing variability with the longer-range parts interpolated from grids of increasing coarseness. Multilevel summation is especially appropriate in the context of dynamics and minimization, because it can produce continuous gradients. This article explores the use of B-splines to increase the accuracy of the multilevel summation method (for nonperiodic boundaries) without incurring additional computation other than a preprocessing step (whose cost also scales linearly). To obtain accurate results efficiently involves technical difficulties, which are overcome by a novel preprocessing algorithm. Numerical experiments demonstrate that the resulting method offers substantial improvements in accuracy and that its performance is competitive with an implementation of the fast multipole method in general and markedly better for Hamiltonian formulations of molecular dynamics. The improvement is great enough to establish multilevel summation as a serious contender for calculating pairwise interactions in molecular dynamics simulations. In particular, the method appears to be uniquely capable for molecular dynamics in two situations, nonperiodic boundary conditions and massively parallel computation, where the fast Fourier transform employed in the particle-mesh Ewald method falls short.


Genotypes and phenotypes of G6PD deficiency among Indonesian females across diagnostic thresholds of G6PD activity guiding safe primaquine therapy of latent malaria.
Plasmodium vivax occurs as a latent infection of liver and a patent infection of red blood cells. Radical cure requires both blood schizontocidal and hypnozoitocidal chemotherapies. The hypnozoitocidal therapies available are primaquine and tafenoquine, 8-aminoquinoline drugs that can provoke threatening acute hemolytic anemia in patients having an X-linked G6PD-deficiency. Heterozygous females may screen as G6PD-normal prior to radical cure and go on to experience hemolytic crisis. This study examined G6PD phenotypes in 1928 female subjects living in malarious Sumba Island in eastern Indonesia to ascertain the prevalence of females vulnerable to diagnostic misclassification as G6PD-normal. All 367 (19%) females having <80% G6PD normal activity were genotyped. Among those, 103 (28%) were G6PD wild type, 251 (68·4%) were heterozygous, three (0·8%) were compound heterozygotes, and ten (2·7%) were homozygous deficient. The variants Vanua Lava, Viangchan, Coimbra, Chatham, and Kaiping occurred among them. Below the 70% of normal G6PD activity threshold, just 18 (8%) were G6PD-normal and 214 (92%) were G6PD-deficient. Among the 31 females with <30% G6PD normal activity were all ten homozygotes, all three compound heterozygotes, and just 18 were heterozygotes (7% of those). In this population, most G6PD heterozygosity in females occurred between 30% and 70% of normal (69·3%; 183/264). The prevalence of females at risk of G6PD misclassification as normal by qualitative screening was 9·5% (183/1928). Qualitative G6PD screening prior to 8-aminoquinoline therapies against P. vivax may leave one in ten females at risk of hemolytic crisis, which may be remedied by point-of-care quantitative tests.


Effects of Lutein and Astaxanthin Intake on the Improvement of Cognitive Functions among Healthy Adults: A Systematic Review of Randomized Controlled Trials.
Fruits and vegetables are generally rich in antioxidants such as carotenoids. Consumption of carotenoids is expected to have benefits on cognitive functions in humans. However, previous randomized controlled trials (RCT) using carotenoids have reported inconsistent results. Therefore, this systematic review (SR) aimed to summarize the effect of carotenoid intake on cognitive functions in humans. PubMed, Cochrane Library, Web of Science, and PsychoINFO were searched for research papers on carotenoid intake with the criteria that 1) oral carotenoid intake was evaluated using RCTs, 2) participants were healthy young, middle-aged, or older, and 3) cognitive functions were measured using RCTs. Five studies using lutein and two studies using astaxanthin met the inclusion criteria. Consumption of lutein and its isomer showed consistent results in selective improvement of visual episodic memory in young and middle-aged adults while inhibition was observed in middle-aged and older adults. One of the two included astaxanthin studies reported a significant improvement of verbal episodic memory performance in middle-aged adults. This SR showed that the 10 mg lutein per day for twelve months can lead to improvement of cognitive functions. Due to the small number of studies, it is difficult to conclude whether astaxanthin would have a positive effect on cognitive functions.


Gambling and Impulsivity Traits: A Recipe for Criminal Behavior?
Gambling disorder (GD) is a psychiatric condition that was recently recategorized as a non-substance-related addiction in the Diagnostic and Statistical Manual of Mental Health Disorders. Criminal activity is commonly associated with gambling; however, few empirical studies to date have examined sociodemographic and psychological variables in this population. In this study, we explored criminal behavior history in a sample of consecutively recruited treatment-seeking gamblers (n = 382) and compared subjects with a history of illegal acts (n = 103, 26.9%) to those with no criminal record (n = 279, 73.1%). Impulsivity and personality traits were specifically explored, along with other gambling-related severity factors. We found that gamblers who engaged in illegal activity were more likely to endorse high levels of urgency (i.e., the tendency to act out when experiencing heightened emotional states) and increased lack of premeditation. Gamblers with a history of criminal behavior also had greater GD severity levels and gambling-related debts. Additionally, these gamblers reported lower levels of self-directedness, which is characterized by difficulty in establishing and redirecting behavior toward one's goals. Likewise, gamblers who had conducted criminal acts showed a tendency to engage in greater risk-taking behavior. These results shed new light on this understudied population and provide insights for developing targeted harm-prevention interventions and treatment protocols.


Gaussian process based nonlinear latent structure discovery in multivariate spike train data.
A large body of recent work focuses on methods for extracting low-dimensional latent structure from multi-neuron spike train data. Most such methods employ either linear latent dynamics or linear mappings from latent space to log spike rates. Here we propose a doubly nonlinear latent variable model that can identify low-dimensional structure underlying apparently high-dimensional spike train data. We introduce the Poisson Gaussian-Process Latent Variable Model (P-GPLVM), which consists of Poisson spiking observations and two underlying Gaussian processes-one governing a temporal latent variable and another governing a set of nonlinear tuning curves. The use of nonlinear tuning curves enables discovery of low-dimensional latent structure even when spike responses exhibit high linear dimensionality (e.g., as found in hippocampal place cell codes). To learn the model from data, we introduce the decoupled Laplace approximation, a fast approximate inference method that allows us to efficiently optimize the latent path while marginalizing over tuning curves. We show that this method outperforms previous Laplace-approximation-based inference methods in both the speed of convergence and accuracy. We apply the model to spike trains recorded from hippocampal place cells and show that it compares favorably to a variety of previous methods for latent structure discovery, including variational auto-encoder (VAE) based methods that parametrize the nonlinear mapping from latent space to spike rates with a deep neural network.


Human polyomavirus related to African green monkey lymphotropic polyomavirus.
While studying the virome of the skin surface of a patient with a Merkel cell carcinoma (MCC) by using unbiased, high-throughput sequencing, we identified a human polyomavirus nearly identical to human polyomavirus 9, a virus recently reported in blood and urine of renal transplantion patients and closely related to the African green monkey lymphotropic polyomavirus. Specific PCR analysis further identified this virus in 2/8 patients with MCC but in only 1/111 controls without MCC. This virus was shed for ≥20 months by the MCC index patient and was on the skin of the spouse of the index patient. These results provide information on the viral ecology of human skin and raise new questions regarding the pathology of virus-associated skin disorders.


Effect of endothelin receptor antagonists on ventricular susceptibility in postinfarcted rats.
This study investigated whether selective endothelin (ET) type A (ET(A)) or nonselective ET(A)/ET(B) receptor blockade exerted antiarrhythmic effects through attenuated sympathetic reinnervation after infarction. Twenty-four hours after ligation of the left anterior descending artery, male Wistar rats received either vehicle, ABT-627 (selective ET(A) receptor antagonist), bosentan (nonselective ET(A)/ET(B) receptor antagonist), or hydralazine for 4 wk. The measurement of myocardial ET-1 levels at the remote zone revealed a significant increase in vehicle-treated infarcted rats compared with sham-operated rats, consistent with increased activities of ET-1 after infarction. Sympathetic nerve function changes assessed by the norepinephrine content of myocardium and the dialysate and plasma dihydroxyphenylglycol levels were parallel to ET-1 levels. Immunohistochemical analysis for tyrosine hydroxylase, growth-associated protein 43, and neurofilament also confirmed the change of nerve function. This was accompanied with a significant upregulation of nerve growth factor protein expression and mRNA in the vehicle-treated infarcted rats, which reduced after the administration of either ET(A) or ET(A)/ET(B) blockade to a similar extent. The beneficial effects of ET receptor antagonists on sympathetic nerve function and structures were dissociated from their blood pressure-lowering effect because ET receptor antagonists and hydralazine reduced arterial pressure similarly. Arrhythmic severity during programmed stimulation in ET receptor antagonists-treated rats was significantly lower than that in vehicle-treated infarcted rats. Our data indicate that the ET system, especially via ET(A) receptors, plays an important role in attenuating sympathetic reinnervation after infarction. Independent of their hemodynamic effects, a chronic use of either ET(A) or ET(A)/ET(B) antagonists may modify the arrhythmogenic response to programmed electrical stimulation.


Effect of a diuretic adjustment algorithm and nonpharmacologic management in patients with heart failure: study protocol for a randomized controlled trial.
One of the challenges in treating patients with heart failure (HF) is achieving clinical stability and reducing the hospital readmission rate. A diuretic dose adjustment algorithm developed in the United States (Diuretic Treatment Algorithm, DTA) and later validated for use in Brazil (as the Algoritmo de Ajuste de Diurético, AAD) has proved feasible and readily applicable, but its effect on clinical outcomes has yet to be assessed. This report aims to describe a randomized clinical trial protocol designed to assess the effectiveness of the AAD and of nonpharmacologic management in improving clinical stability and reducing the readmission rate at 90 days in patients with HF. A PROBE (prospective randomized open blinded endpoint) parallel-group design will be used. Adult patients with a diagnosis of reduced ejection fraction HF, who are being treated at a specialized HF clinic are being recruited. Those with indications for loop diuretic dose adjustment during routine clinic visits will be randomized to take part in the trial. Participants in the intervention group (IG) shall have their diuretic doses adjusted in accordance with the AAD and receive four telephone calls (one per week) over 30 days to reinforce guidance on nonpharmacological management (fluid and sodium restriction). Participants in the control group (CG) shall have their diuretic doses adjusted by a physician during the first trial visit and shall not receive any telephone calls. Patients in both groups shall return at 1 month for face-to-face reassessment. The study endpoints shall comprise readmission and/or emergency department visits due to HF decompensation within 90 days and clinical instability. All participants shall be required to have a scale at home (or easy access to one), a telephone number, agree to telephone-based follow-up, and be available to return for a 1-month trial visit. Overall, 135 patients are expected to be enrolled in each group. This trial shall assess the effectiveness of the AAD algorithm and non-pharmacologic management by early identification of clinical deterioration and establishment of a combined intervention to reduce emergency department visits, readmission rate, or a composite endpoint thereof. ClinicalTrials.gov Identifier, NCT02068937 (23 February 2014).


Prioritization of unregulated disinfection by-products in drinking water distribution systems for human health risk mitigation: A critical review.
Water disinfection involves the use of different types of disinfectants, which are oxidizing agents that react with natural organic matter (NOM) to form disinfection by-products (DBPs). The United States Environmental Protection Agency (USEPA) has established threshold limits on some DBPs, which are known as regulated DBPs (R-DBPs). The human health risks associated with R-DBPs in drinking water distribution systems (DWDSs) and application of stricter regulations have led water utilities to switch from conventional disinfectant (i.e., chlorination) to alternative disinfectants. However, the use of alternative disinfectants causes formation of a new suit of DBPs known as unregulated DBPs (UR-DBPs), which in many cases can be more toxic. There is a growing concern of UR-DBPs formation in drinking water. This review prioritizes some commonly occurring UR-DBP groups and species in DWDSs based on their concentration level, reported frequency, and toxicity using an indexing method. There are nine UR-DBPs group and 36 species that have been identified based on recent published peer-reviewed articles. Haloacetonitriles (HANs) and haloacetaldehydes (HALs) are identified as important UR-DBP groups. Dichloroacetonitrile (DCAN) and trichloroacetaldehye (TCAL) are identified as critical UR-DBPs species. The outcomes of this review can help water regulators to identify the most critical UR-DBPs species in the context of drinking water safety and provide them with useful information to develop guidelines or threshold limits for UR-DBPs. The outcomes can also help water utilities in selecting water treatment processes for the mitigation of human health risk posed by UR-DBPs through drinking water.


Domestic horses (Equus caballus) prefer to approach humans displaying a submissive body posture rather than a dominant body posture.
Signals of dominance and submissiveness are central to conspecific communication in many species. For domestic animals, sensitivities to these signals in humans may also be beneficial. We presented domestic horses with a free choice between two unfamiliar humans, one adopting a submissive and the other a dominant body posture, with vocal and facial cues absent. Horses had previously been given food rewards by both human demonstrators, adopting neutral postures, to encourage approach behaviour. Across four counterbalanced test trials, horses showed a significant preference for approaching the submissive posture in both the first trial and across subsequent trials, and no individual subject showed an overall preference for dominant postures. There was no significant difference in latency to approach the two postures. This study provides novel evidence that domestic horses may spontaneously discriminate between, and attribute communicative significance to, human body postures of dominance; and further, that familiarity with the signaller is not a requirement for this response. These findings raise interesting questions about the plasticity of social signal perception across the species barrier.


Estimating the Population Sizes of Men Who Have Sex With Men in US States and Counties Using Data From the American Community Survey.
In the United States, male-to-male sexual transmission accounts for the greatest number of new human immunodeficiency virus (HIV) diagnoses and a substantial number of sexually transmitted infections (STI) annually. However, the prevalence and annual incidence of HIV and other STIs among men who have sex with men (MSM) cannot be estimated in local contexts because demographic data on sexual behavior, particularly same-sex behavior, are not routinely collected by large-scale surveys that allow analysis at state, county, or finer levels, such as the US decennial census or the American Community Survey (ACS). Therefore, techniques for indirectly estimating population sizes of MSM are necessary to supply denominators for rates at various geographic levels. Our objectives were to indirectly estimate MSM population sizes at the county level to incorporate recent data estimates and to aggregate county-level estimates to states and core-based statistical areas (CBSAs). We used data from the ACS to calculate a weight for each county in the United States based on its relative proportion of households that were headed by a male who lived with a male partner, compared with the overall proportion among counties at the same level of urbanicity (ie, large central metropolitan county, large fringe metropolitan county, medium/small metropolitan county, or nonmetropolitan county). We then used this weight to adjust the urbanicity-stratified percentage of adult men who had sex with a man in the past year, according to estimates derived from the National Health and Nutrition Examination Survey (NHANES), for each county. We multiplied the weighted percentages by the number of adult men in each county to estimate its number of MSM, summing county-level estimates to create state- and CBSA-level estimates. Finally, we scaled our estimated MSM population sizes to a meta-analytic estimate of the percentage of US MSM in the past 5 years (3.9%). We found that the percentage of MSM among adult men ranged from 1.5% (Wyoming) to 6.0% (Rhode Island) among states. Over one-quarter of MSM in the United States resided in 1 of 13 counties. Among counties with over 300,000 residents, the five highest county-level percentages of MSM were San Francisco County, California at 18.5% (66,586/359,566); New York County, New York at 13.8% (87,556/635,847); Denver County, Colorado at 10.5% (25,465/243,002); Multnomah County, Oregon at 9.9% (28,949/292,450); and Suffolk County, Massachusetts at 9.1% (26,338/289,634). Although California (n=792,750) and Los Angeles County (n=251,521) had the largest MSM populations of states and counties, respectively, the New York City-Newark-Jersey City CBSA had the most MSM of all CBSAs (n=397,399). We used a new method to generate small-area estimates of MSM populations, incorporating prior work, recent data, and urbanicity-specific parameters. We also used an imputation approach to estimate MSM in rural areas, where same-sex sexual behavior may be underreported. Our approach yielded estimates of MSM population sizes within states, counties, and metropolitan areas in the United States, which provide denominators for calculation of HIV and STI prevalence and incidence at those geographic levels.


Exerciser for rehabilitation of the Arm (ERA): Development and unique features of a 3D end-effector robot.
Stroke is one of the leading causes of disability worldwide. Consequently, many stroke survivors exhibit difficulties undergoing voluntary movement in their affected upper limb, compromising their functional performance and level of independence. To minimize the negative impact of stroke disabilities, exercises are recognized as a key element in post-stroke rehabilitation. In order to provide the practice of exercises in a uniform and controlled manner as well as increasing the efficiency of therapists' interventions, robotic training has been found, and continues to prove itself, as an innovative intervention for post-stroke rehabilitation. However, the complexity as well as the limited degrees of freedom and workspace of currently commercially available robots can limit their use in clinical settings. Up to now, user-friendly robots covering a sufficiently large workspace for training of the upper limb in its full range of motion are lacking. This paper presents the design and implementation of ERA, an upper-limb 3-DOF force-controlled exerciser robot, which presents a workspace covering the entire range of motion of the upper limb. The ERA robot provides 3D reaching movements in a haptic virtual environment. A description of the hardware and software components of the ERA robot is also presented along with a demonstration of its capabilities in one of the three operational modes that were developed.


Retention force and allowable range of the angle of an implant-supported overdenture attachment system using healing screws and a silicone resilient denture liner.
Changes in the intraoral condition immediately after implant placement and in patients using fixed prosthetic appliances with implant abutments cause problems that require the use of an easily adjustable implant-supported overdenture system. The purpose of this in vitro study was to develop a simulated implant-supported overdenture attachment system by using healing screws for the patrix and a silicone resilient denture liner for the matrix and to investigate the initial retention force, time-course changes, and allowable range of the angle between attachments. Tests on the retention force and allowable range of the angle were performed. Attachments using tissue-level healing screws (height: 2.0 and 3.0 mm) for the patrix and a silicone resilient denture liner for the matrix were prepared. In the retention force measurement test, the frequency of insertion and removal was set at 3 per day to simulate a 4-month relief period. The joined attachment model was pulled apart, and the maximum traction (N) required to remove it was defined as the retention force. The retention force was measured every 90 times (representing the number of insertions and removals per month). To test the allowable angle range, 2 patrices were used. The angle between the 2 patrices was set at 0, 10, 20, and 30 degrees, and the angular limit for joining with the matrix was measured. The initial retention force of the healing screw attachments was compared with that of a polymeric O-ring by using 1-way layout ANOVA followed by the Bonferroni test (α=.05). To analyze time-course changes in the 2 types of healing screw attachments, the retention force before insertion and removal was compared with that after repeated insertions and removals for each simulated period using 1-way layout ANOVA followed by the Dunnett test (α=.05). The initial retention force of the 2.0- and 3.0-mm healing screws was 2.4 ±0.1 and 2.6 ±0.2 N. After repeating insertions and removals to simulate use for 4 months, the retention force of the 2.0- and 3.0-mm healing screws was 1.8 ±0.2 and 2.2 ±0.1 N, respectively, both showing significant differences from the initial retention force (P<.05). The allowable angle range test revealed that insertion and removal of the healing screws was possible up to 30 degrees. Under the conditions of this in vitro study, repeated insertion and removal attenuated the retention force but was still equivalent to the retention force of the O-ring. The allowable range of the angle between patrices for insertion and removal was up to 30 degrees.


Analysis of genetic linkage data for Mendelian traits.
This unit describes linkage analysis, an invaluable tool in mapping human disease genes. Linkage analysis is one of several methods used to map genes. Specifically, in linkage analysis, cosegregation of two or more genes (traits) is examined in a family unit to determine if they segregate independently of each other according to Mendel's laws or if they do not segregate independently because of their close physical proximity. The procedures outlined in this unit are not always straightforward, and a number of caveats are presented in the commentary regarding possible complications and confounding factors that may arise. Specific examples are given, but it is not possible to cover all possible scenarios or variables. The intention is to enable the reader to master basic principles used in the linkage approach, and thus be in a position to undertake, with proper consultation, a linkage-analysis study. This unit deals specifically with lod score analysis in Mendelian disorders.


Endometrium and steroids, a pathologic overview.
Normal endometrial function requires of cell proliferation and differentiation; therefore, disturbances in these processes could lead to pathological entities such as hyperplasia and endometrial adenocarcinoma, where cell proliferation is increased. The development of these pathologies is highly related to alterations in the levels and/or action of sexual steroids. In the present review, it has been analyzed how steroids, particularly estrogens, androgens and progestagens are involved in the etiopathogenesis of hyperplasia and endometrial endometrioid adenocarcinoma. The emphasis is given on pathological and pharmacological conditions that are presented as risk factors for endometrial pathologies, such as obesity, polycystic ovarian syndrome and hormone replacement postmenopausal women therapy, among others. Steroids alterations may promote changes at molecular level that enhance the development of hyperplasia and endometrioid cancer. In fact, there are solid data that indicate that estrogens stimulate cell-proliferation in this tissue; meanwhile, progestagens are able to stop cell proliferation and to increase differentiation. Nevertheless, the role of androgens is less clear, since there is contradictory information. It is most likely that the major contribution of steroids to the development of cell proliferation pathologies in endometria would be in early stages, where there is a high sensitivity to these molecules. This phenomenon is present even in stages previous to the occurrence of hyperplasia, like in the condition of polycystic ovarian syndrome, where the endometria have a greater sensitivity to steroids and high expression of cell cycle molecules. These abnormalities would contribute to the pathogenesis of hyperplasia and then in the progression to endometrioid adenocarcinoma.


STRING: known and predicted protein-protein associations, integrated and transferred across organisms.
A full description of a protein's function requires knowledge of all partner proteins with which it specifically associates. From a functional perspective, 'association' can mean direct physical binding, but can also mean indirect interaction such as participation in the same metabolic pathway or cellular process. Currently, information about protein association is scattered over a wide variety of resources and model organisms. STRING aims to simplify access to this information by providing a comprehensive, yet quality-controlled collection of protein-protein associations for a large number of organisms. The associations are derived from high-throughput experimental data, from the mining of databases and literature, and from predictions based on genomic context analysis. STRING integrates and ranks these associations by benchmarking them against a common reference set, and presents evidence in a consistent and intuitive web interface. Importantly, the associations are extended beyond the organism in which they were originally described, by automatic transfer to orthologous protein pairs in other organisms, where applicable. STRING currently holds 730,000 proteins in 180 fully sequenced organisms, and is available at http://string.embl.de/.


Molecular diagnosis of suspected tuberculosis from archived smear slides from the Balimo region, Papua New Guinea.
Tuberculosis (TB) is a serious health problem in Papua New Guinea (PNG) with an estimated 30000 new cases and 3800 deaths each year. In the Balimo region of the Western Province, diagnosis relies on clinical manifestations and on the microscopic detection of acid-fast bacilli (AFB) in sputum smears, a technique with limited sensitivity. A molecular diagnosis assay targeting DNA extracted from archived sputum smear slides collected from the Balimo region (2012-2014) was conducted, without the need for a viable culture. The presence of Mycobacterium sp on 1162 slides prepared from 345 sputum samples was assessed using a real-time PCR (qPCR) approach. The qPCR technique identified the presence of mycobacteria in 35.4% of the smear slides and 59.7% of the tested sputum samples. Poor agreement was observed between the two diagnosis methods (smear AFB microscopy versus qPCR), with 100 AFB-positive sputum samples compared to 206 qPCR-positive sputum samples overall. Treatment was initiated in 90.2% of the smear-positive cases. Unnecessary treatment of 'false-positive' TB cases (AFB-negative/qPCR-negative) was very low (8.6%) and was even lower when the nine patients diagnosed with extrapulmonary TB were excluded from the analysis. However, the prevalence of false-negatives (AFB-negative/qPCR-positive) was high (28.5%). Undetected smear-negative TB is occurring in the Balimo region of PNG, as well as some unnecessary empirical treatment. Molecular methods of diagnosis could greatly reduce the frequency of inappropriate clinical assessment, as well as providing point-of-care diagnosis. This may provide substantial patient and programmatic benefits, including lowering the economic burden on patients from rural areas seeking medical diagnosis in Balimo.


Tocopherol (vitamin E) in Alzheimer's disease and other neurodegenerative disorders.
In this article, we review the evidence that tocopherol (vitamin E) may have a role to play in the prevention and treatment of Alzheimer's disease and other neurological diseases. The theoretical rationale for the effectiveness of tocopherol as treatment and/or prevention of Alzheimer's disease is based on its antioxidant properties. Results from animal and in vitro studies provide evidence to support use of tocopherol for prevention and treatment of degenerative neurological diseases. Furthermore, several, but not all, epidemiological, cross-sectional, prospective studies indicate that tocopherol may have protective effects in Alzheimer's disease, although dietary and supplemental forms of the vitamin may differ in their efficacy. Mixed results have been obtained from clinical trials. Evidence of the use of tocopherol as a protective measure or as therapy in neurological diseases other than Alzheimer's disease is less compelling. To date, there are no clear-cut answers as to whether tocopherol is worth prescribing, but current clinical practice favours its use in the treatment of Alzheimer's disease.


Statin use and tooth loss in chronic periodontitis patients.
Statins have anti-inflammatory and bone stimulating properties that may positively affect chronic periodontitis. Our objective in this study was to evaluate whether statin use by chronic periodontitis patients had a beneficial impact on tooth loss. In a retrospective cohort study (N patients = 1,021; mean follow-up = 7.1 years), dental records were merged with pharmacy data. Any statin use during 3 years, statin use during each of 3 consecutive years (regular use), and any statin use during the first 3 years after the initial periodontal exam were evaluated as predictors of tooth loss using negative binomial regression models with adjustment for potential confounding factors. Any statin use during 3 years was not associated with tooth loss rate in the year subsequent to the 3-year period (rate ratio [RR] = 1.00; 95% confidence interval [CI] = 0.56 to 1.81). Regular statin use during 3 years was associated with a non-significant 37% reduced tooth loss rate in the year subsequent to the 3-year period (RR = 0.63; 95% CI = 0.32 to 1.25). Any statin use during the first 3 years after the initial periodontal exam was associated with a 48% decreased tooth loss rate in year 4 and subsequent years (RR = 0.52; 95% CI = 0.29 to 0.95). Our findings were mixed for an association of statin use with reduced tooth loss in chronic periodontitis patients. Lack of control for some potential confounders, particularly smoking, and evaluation of different patterns of statin usage hamper the interpretation of the results. Exploration of these findings in additional epidemiological studies may be worthwhile.


Role of Bicaudal-D in patterning the Drosophila egg chamber in mid-oogenesis.
The Bicaudal-D (Bic-D) gene is required early in Drosophila oogenesis for the differentiation of an oocyte from one of a cluster of 16 interconnected germarial cells. To analyze the role of Bic-D later in oogenesis, we have constructed Drosophila lines in which Bic-D expression is under the control of the hsp70 promoter. In these flies, Bic-D activity can be induced early in oogenesis, allowing an oocyte to be made. Then, by shifting females to non-inducing conditions, Bic-D levels are depleted for the remainder of oogenesis. Using this system, we find that Bic-D is indeed required in the later stages of oogenesis. In ovaries from mutant females, oocyte growth is reduced, apparently due to defects in nurse-cell-to-oocyte transport. Smaller oocyte size results in the misalignment of follicle cells and the underlying germ line, leading to ventralization of dorsal follicle cells and to defects in centripetal cell migration. In addition, we show that Bic-D is required for the localization of specific mRNAs at both the anterior and posterior of the oocyte.


A randomized comparison of cardiocerebral and cardiopulmonary resuscitation using a swine model of prolonged ventricular fibrillation.
Cardiocerebral resuscitation (CCR) is reportedly superior to cardiopulmonary resuscitation (CPR) for primary cardiac arrest in the prehospital setting. This study was done using a swine model of prolonged ventricular fibrillation (VF) to quantify the effect of the emergency medical services component of CCR with intraosseous access (CCR-IO) compared with standard CPR with intravenous access (CPR-IV) as it is typically performed during out-of-hospital cardiac arrest (OHCA) resuscitation in a prospective randomized fashion. Fifty-three animals were instrumented under anesthesia and VF was electrically induced. After 10 min of untreated VF, baseline characteristics were recorded, and animals were block randomized to one of two resuscitation schemes. The controls had mechanical chest compressions at 100/min with ventilations at a ratio of 30:2. Consistent with clinical practice, two 30-s pauses in chest compressions occurred to simulate attempts to accomplish endotracheal intubation at minutes 1 and 3 of CPR and successful IV access was simulated to occur three additional minutes after endotracheal intubation. The CCR group had continuous uninterrupted mechanical chest compressions at 100/min. No active ventilations were provided. A tibial IO needle was placed in real time for vascular access. Both groups received epinephrine (0.1 mg/kg) as soon as access became available followed by 2.5 min of chest compressions before the first 120 J rescue shock attempt. After successful rescue shock, standardized post-resuscitative care was provided to a 20-min endpoint. Failed rescue shock was followed by continued chest compressions with positive pressure ventilation in both groups, repeat doses of epinephrine (0.01 mg/kg) every 3 min, and rescue shock every minute as long as a shockable rhythm persisted. Group comparisons were assessed using descriptive statistics. Proportions with 95% confidence intervals were calculated for VF termination, ROSC, and survival. Baseline characteristics and chemistries between the two groups at VF induction and after 10 min of non-treatment were mathematically the same. The proportions of VF termination (0.50 vs. 0.82), ROSC (0.30 vs. 0.59), and 20-min survival (0.19 vs. 0.40) all strongly favored the CCR-IO group. In this swine model of witnessed VF arrest with no bystander-initiated resuscitation, CCR-IO resulted in substantial improvement in all three outcomes relative to typical emergency medical services provided CPR-IV.


Three-line fluorescence thermometry of optically thick shock-tunnel flow.
Two-line fluorescence thermometry measurements based on excitation of the A-X (0, 0) band of nitric oxide were performed in the free-stream flow of a large-scale shock tunnel, the Arnold Engineering Development Center Impulse Facility. Because the flow is optically thick, effects of laser absorption must be taken into account. This can be accomplished by extending the two-line thermometry concept to three lines. We illustrate the three-line technique by combining two-line planar laser-induced fluorescence measurements from separate facility runs. The resulting free-stream temperature T(0) = 468 K ? 65 K is insensitive to assumptions regarding the laser propagation distance through the flow, the vibrational temperature of the flow, and the precise nature of the absorption line shape.


Carbohydrate-attached fullerene derivative for selective localization in ordered carbohydrate-block-poly(3-hexylthiophene) nanodomains.
A carbohydrate-based fullerene derivative (AcMal7-C61) is designed, synthesized and applied to a lamellar-forming high-χ block copolymer system, poly(3-hexylthiophene)-block-peracetylated maltoheptaose (P3HT-b-AcMal7), to actualize an ordered donor/acceptor (D/A) network. A well-defined D/A lamellar structure of the P3HT-b-AcMal7:AcMal7-C61 blend with sub-10 nm domain features is achieved upon thermal annealing. The AcMal7-C61 molecules are localized in the phase-separated AcMal7 nanodomains without causing the formation of fullerene crystals while maintaining the lamellar morphology up to 1:0.5 (D:A) blending ratio. The cross-sectional TEM observation and GISAXS measurement reveals that the P3HT-b-AcMal7 tends to spontaneously organize into lamellar structures oriented perpendicular to the film surface at the air/film interface while the domain orientation at the bottom interface depends on the nature of the substrate.


Electric field gradients in Hg compounds: molecular orbital (MO) analysis and comparison of 4-component and 2-component (ZORA) methods.
We examine the performance of Density Functional Theory (DFT) approaches based on the Zeroth-Order Regular Approximation (ZORA) Hamiltonian (with and without inclusion of spin-orbit coupling) for predictions of electric field gradients (EFGs) at the heavy atom Hg nucleus. This is achieved by comparing with benchmark DFT and CCSD-T data (Arcisauskaite et al., Phys. Chem. Chem. Phys., 2012, 14, 2651-2657) obtained from 4-component Dirac-Coulomb Hamiltonian calculations. The investigated set of molecules comprises linear HgL(2) (L = Cl, Br, I, CH(3)) and bent HgCl(2) mercury compounds as well as the trigonal planar [HgCl(3)](-) system. In 4-component calculations we used the dyall.cv3z basis set for Hg, Br, I and the cc-pCVTZ basis set for H, C, Cl, whereas in ZORA calculations we used the QZ4P basis set for all the atoms. ZORA-4 reproduces the fully relativistic 4-component DFT reference values within 6% for all studied Hg compounds and employed functionals (BH&H, BP86, PBE0), whereas scalar relativistic (SR)-ZORA-4 results show deviations of up to 15%. Compared to our 4-component CCSD-T benchmark the BH&H functional performs best at both 4-component and ZORA levels. We furthermore observe that changes in the largest component of the diagonalised EFG tensor, V(zz), of linear HgCl(2) show a slightly stronger dependence than the r(-3) scaling upon bond length r(Hg-Cl) alterations. The 4-component/BH&H V(zz) value of -9.26 a.u. for a bent HgCl(2) (∠Cl-Hg-Cl = 120°) is close to -9.60 a.u. obtained for the linear HgCl(2) structure. Thus a point charge model for EFG calculations completely fails in this case. By means of a projection analysis of molecular orbital (MO) contributions to V(zz) in terms of the atomic constituents, we conclude that this is due to the increased importance of the Hg 5d orbitals upon bending HgCl(2) compared to the linear HgCl(2) structure. Changing ligand leads to only minor changes in V(zz) (from -9.60 a.u. (HgCl(2)) to -8.85 a.u. (HgI(2)) at the 4-component/BH&H level). This appears to be due to cancellation of contributions with opposite signs to V(zz) arising from: (i) increasing electron donation from occupied ligand orbitals to the formally empty Hg 6p orbitals and (ii) an increasing bond length and a decreasing negative charge on the ligand along the series.


Contribution of horizontal gene transfer to the evolution of Saccharomyces cerevisiae.
The genomes of the hemiascomycetes Saccharomyces cerevisiae and Ashbya gossypii have been completely sequenced, allowing a comparative analysis of these two genomes, which reveals that a small number of genes appear to have entered these genomes as a result of horizontal gene transfer from bacterial sources. One potential case of horizontal gene transfer in A. gossypii and 10 potential cases in S. cerevisiae were identified, of which two were investigated further. One gene, encoding the enzyme dihydroorotate dehydrogenase (DHOD), is potentially a case of horizontal gene transfer, as shown by sequencing of this gene from additional bacterial and fungal species to generate sufficient data to construct a well-supported phylogeny. The DHOD-encoding gene found in S. cerevisiae, URA1 (YKL216W), appears to have entered the Saccharomycetaceae after the divergence of the S. cerevisiae lineage from the Candida albicans lineage and possibly since the divergence from the A. gossypii lineage. This gene appears to have come from the Lactobacillales, and following its acquisition the endogenous eukaryotic DHOD gene was lost. It was also shown that the bacterially derived horizontally transferred DHOD is required for anaerobic synthesis of uracil in S. cerevisiae. The other gene discussed in detail is BDS1, an aryl- and alkyl-sulfatase gene of bacterial origin that we have shown allows utilization of sulfate from several organic sources. Among the eukaryotes, this gene is found in S. cerevisiae and Saccharomyces bayanus and appears to derive from the alpha-proteobacteria.


The RANK/RANK ligand system is involved in interleukin-6 and interleukin-11 up-regulation by human myeloma cells in the bone marrow microenvironment.
The receptor activator of NF-kB ligand (RANKL) has a critical role in osteoclast activation. Recently it has been demonstrated that human multiple myeloma (MM) cells do not express RANKL but up-regulate RANKL in bone marrow stromal cells (BMSC). To further investigate the role of RANKL in the pathophysiology of MM we evaluated the expression of its receptor RANK in MM cells and in the BM environment and the potential role of RANKL in the interaction of myeloma cells with the microenvironment. RANK mRNA and protein expression were evaluated by reverse transcription polymerase chain reaction and Western blot analysis in human myeloma cell lines (HMCL), fresh purified MM cells, BMSC and endothelial cells. Moreover the effect and the role of RANKL on cytokine secretion were evaluated in BMSC, in endothelial cells and in co-culture conditions with myeloma cells. We found that RANK is expressed in BMSC and endothelial cells but not in myeloma cells. Consistently, RANKL did not have a direct effect on myeloma cell survival, but RANKL treatment induced a significant increase of interleukin (IL)-6 and IL-11 secretion by both BMSC and endothelial cells. Moreover, in a co-culture system we found that myeloma cells up-regulated both IL-6 and IL-11 secretion by BMSC and endothelial cells through cell-to-cell contact. The presence of the RANK-Fc that blocks the RANK/RANKL interaction significantly inhibited HMCL-induced secretion of IL-6 and IL-11. Our data provide new notions on the role of the RANKL system in the pathophysiology of MM.


Cellular localization of cytochrome c550. Its specific association with cyanobacterial photosystem II.
Cellular localization of cytochrome (cyt) c550, a low potential, c-type monoheme cytochrome, in a thermophilic cyanobacterium Synechococcus vulcanus was investigated by systematic fractionation of the cells followed by its enzymatic and immunological detection. While cyt c-553, a soluble cyt that donates an electron to P700, was detected in the supernatant of osmotic disruption of lysozyme-treated cells, cyt c550 was detected only in the thylakoid membrane fraction, being tightly bound to thylakoids, and its removal required sonication in the presence of 1 M CaCl2. Upon further fractionation of the thylakoids into photosystem (PS) I and PSII by lauryl dimethylamine N-oxide solubilization, cyt c550 was exclusively concentrated in the crude PSII fraction together with cyt f, with no significant amount being detected in any of the soluble and PSI fractions. Upon further fractionation of the crude PSII by n-dodecyl beta-D-maltoside solubilization followed by column chromatography, cyt c550 was detected exclusively in the purified PSII core complex fraction but not in any other fractions. A 12-kDa protein, one of the extrinsic components of cyanobacterial PSII, exhibited completely the same behavior as that of cyt c550 during these fractionation procedures. These results, coupled with our previous results that cyt c550 binds stoichiometrically to the cyanobacterial PSII core complex and enhances O2 evolution (Shen, J.-R., and Inoue, Y. (1993) Biochemistry 32, 1825-1832), indicate that there is only one species of cyt c550 in cyanobacterial cells and that this cyt is exclusively associated with PSII as a functional component for O2 evolution.


Artemis is a phosphorylation target of ATM and ATR and is involved in the G2/M DNA damage checkpoint response.
Mutations in Artemis in both humans and mice result in severe combined immunodeficiency due to a defect in V(D)J recombination. In addition, Artemis mutants are radiosensitive and chromosomally unstable, which has been attributed to a defect in nonhomologous end joining (NHEJ). We show here, however, that Artemis-depleted cell extracts are not defective in NHEJ and that Artemis-deficient cells have normal repair kinetics of double-strand breaks after exposure to ionizing radiation (IR). Artemis is shown, however, to interact with known cell cycle checkpoint proteins and to be a phosphorylation target of the checkpoint kinase ATM or ATR after exposure of cells to IR or UV irradiation, respectively. Consistent with these findings, our results also show that Artemis is required for the maintenance of a normal DNA damage-induced G2/M cell cycle arrest. Artemis does not appear, however, to act either upstream or downstream of checkpoint kinase Chk1 or Chk2. These results define Artemis as having a checkpoint function and suggest that the radiosensitivity and chromosomal instability of Artemis-deficient cells may be due to defects in cell cycle responses after DNA damage.


Child abuse: A classic case report with literature review.
Child abuse and neglect are serious global problems and can be in the form of physical, sexual, emotional or just neglect in providing for the child's needs. These factors can leave the child with serious, long-lasting psychological damage. In the present case report, a 12-year-old orphaned boy was physically abused by a close relative who caused actual bodily and emotional trauma to the boy. After satisfactorily managing the trauma and emotional effects to the patient, in addition to the counseling services provided to the caregiver, the patient made a steady recovery. He was also referred to a child support group for social support, and prepare him together with his siblings for placement in a children's home in view of the hostile environment in which they were living.


Global self-organization of the cellular metabolic structure.
Over many years, it has been assumed that enzymes work either in an isolated way, or organized in small catalytic groups. Several studies performed using "metabolic networks models" are helping to understand the degree of functional complexity that characterizes enzymatic dynamic systems. In a previous work, we used "dissipative metabolic networks" (DMNs) to show that enzymes can present a self-organized global functional structure, in which several sets of enzymes are always in an active state, whereas the rest of molecular catalytic sets exhibit dynamics of on-off changing states. We suggested that this kind of global metabolic dynamics might be a genuine and universal functional configuration of the cellular metabolic structure, common to all living cells. Later, a different group has shown experimentally that this kind of functional structure does, indeed, exist in several microorganisms. Here we have analyzed around 2.500.000 different DMNs in order to investigate the underlying mechanism of this dynamic global configuration. The numerical analyses that we have performed show that this global configuration is an emergent property inherent to the cellular metabolic dynamics. Concretely, we have found that the existence of a high number of enzymatic subsystems belonging to the DMNs is the fundamental element for the spontaneous emergence of a functional reactive structure characterized by a metabolic core formed by several sets of enzymes always in an active state. This self-organized dynamic structure seems to be an intrinsic characteristic of metabolism, common to all living cellular organisms. To better understand cellular functionality, it will be crucial to structurally characterize these enzymatic self-organized global structures.


Immunosuppressive activity enhances central carbon metabolism and bioenergetics in myeloid-derived suppressor cells in vitro models.
The tumor microenvironment contains a vast array of pro- and anti-inflammatory cytokines that alter myelopoiesis and lead to the maturation of immunosuppressive cells known as myeloid-derived suppressor cells (MDSCs). Incubating bone marrow (BM) precursors with a combination of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-6 (IL-6) generated a tumor-infiltrating MDSC-like population that impaired anti-tumor specific T-cell functions. This in vitro experimental approach was used to simulate MDSC maturation, and the cellular metabolic response was then monitored. A complementary experimental model that inhibited L-arginine (L-Arg) metabolizing enzymes in MSC-1 cells, an immortalized cell line derived from primary MDSCs, was used to study the metabolic events related to immunosuppression. Exposure of BM cells to GM-CSF and IL-6 activated, within 24 h, L-Arg metabolizing enzymes which are responsible for the MDSCs immunosuppressive potential. This was accompanied by an increased uptake of L-glutamine (L-Gln) and glucose, the latter being metabolized by anaerobic glycolysis. The up-regulation of nutrient uptake lead to the accumulation of TCA cycle intermediates and lactate as well as the endogenous synthesis of L-Arg and the production of energy-rich nucleotides. Moreover, inhibition of L-Arg metabolism in MSC-1 cells down-regulated central carbon metabolism activity, including glycolysis, glutaminolysis and TCA cycle activity, and led to a deterioration of cell bioenergetic status. The simultaneous increase of cell specific concentrations of ATP and a decrease in ATP-to-ADP ratio in BM-derived MDSCs suggested cells were metabolically active during maturation. Moreover, AMP-activated protein kinase (AMPK) was activated during MDSC maturation in GM-CSF and IL-6-treated cultures, as revealed by the continuous increase of AMP-to-ATP ratios and the phosphorylation of AMPK. Likewise, AMPK activity was decreased in MSC-1 cells when L-Arg metabolizing enzymes were inhibited. Finally, inhibition of AMPK activity by the specific inhibitor Compound C (Comp-C) resulted in the inhibition of L-Arg metabolizing enzyme activity and abolished MDSCs immunosuppressive activity. We anticipate that the inhibition of AMPK and the control of metabolic fluxes may be considered as a novel therapeutic target for the recovery of the immunosurveillance process in cancer-bearing hosts.


Quantification of Transcriptome Responses of the Rumen Epithelium to Butyrate Infusion using RNA-seq Technology.
Short-chain fatty acids (SCFAs), such as butyrate, produced by gut microorganisms, play a critical role in energy metabolism and physiology of ruminants as well as in human health. In this study, the temporal effect of elevated butyrate concentrations on the transcriptome of the rumen epithelium was quantified via serial biopsy sampling using RNA-seq technology. The mean number of genes transcribed in the rumen epithelial transcriptome was 17,323.63 ± 277.20 (±SD; N = 24) while the core transcriptome consisted of 15,025 genes. Collectively, 80 genes were identified as being significantly impacted by butyrate infusion across all time points sampled. Maximal transcriptional effect of butyrate on the rumen epithelium was observed at the 72-h infusion when the abundance of 58 genes was altered. The initial reaction of the rumen epithelium to elevated exogenous butyrate may represent a stress response as Gene Ontology (GO) terms identified were predominantly related to responses to bacteria and biotic stimuli. An algorithm for the reconstruction of accurate cellular networks (ARACNE) inferred regulatory gene networks with 113,738 direct interactions in the butyrate-epithelium interactome using a combined cutoff of an error tolerance (ɛ = 0.10) and a stringent P-value threshold of mutual information (5.0 × 10(-11)). Several regulatory networks were controlled by transcription factors, such as CREBBP and TTF2, which were regulated by butyrate. Our findings provide insight into the regulation of butyrate transport and metabolism in the rumen epithelium, which will guide our future efforts in exploiting potential beneficial effect of butyrate in animal well-being and human health.


Automated mass spectrometric analysis of urinary free catecholamines using on-line solid phase extraction.
Analysis of catecholamines (epinephrine, norepinephrine and dopamine) in plasma and urine is used for diagnosis and treatment of catecholamine-producing tumors. Current analytical techniques for catecholamine quantification are laborious, time-consuming and technically demanding. Our aim was to develop an automated on-line solid phase extraction method coupled to high performance liquid chromatography-tandem mass spectrometry (XLC-MS/MS) for the quantification of free catecholamines in urine. Five microlitre urine equivalent was pre-purified by automated on-line solid phase extraction, using phenylboronic acid complexation. Reversed phase (pentafluorophenylpropyl column) chromatography was applied. Mass spectrometric detection was operated in multiple reaction monitoring mode using a quadrupole tandem mass spectrometer with positive electrospray ionization. Urinary reference intervals were set in 24-h urine collections of 120 healthy subjects. XLC-MS/MS was compared with liquid chromatography with electrochemical detection (HPLC-ECD). Total run-time was 14 min. Intra- and inter-assay analytical variations were <10%. Linearity was excellent (R2>0.99). Quantification limits were 1.47 nmol/L, 15.8 nmol/L and 11.7 nmol/L for epinephrine, norepinephrine and dopamine, respectively. XLC-MS/MS correlated well with HPLC-ECD (correlation coefficient >0.98). Reference intervals were 1-10 micromol/mol, 10-50 micromol/mol and 60-225 micromol/mol creatinine for epinephrine, norepinephrine and dopamine, respectively. Advantages of the XLC-MS/MS catecholamine method include its high analytical performance by selective PBA affinity and high specificity and sensitivity by unique MS/MS fragmentation.


Sliding sleeves of XRCC4-XLF bridge DNA and connect fragments of broken DNA.
Non-homologous end joining (NHEJ) is the primary pathway for repairing DNA double-strand breaks (DSBs) in mammalian cells. Such breaks are formed, for example, during gene-segment rearrangements in the adaptive immune system or by cancer therapeutic agents. Although the core components of the NHEJ machinery are known, it has remained difficult to assess the specific roles of these components and the dynamics of bringing and holding the fragments of broken DNA together. The structurally similar XRCC4 and XLF proteins are proposed to assemble as highly dynamic filaments at (or near) DSBs. Here we show, using dual- and quadruple-trap optical tweezers combined with fluorescence microscopy, how human XRCC4, XLF and XRCC4-XLF complexes interact with DNA in real time. We find that XLF stimulates the binding of XRCC4 to DNA, forming heteromeric complexes that diffuse swiftly along the DNA. Moreover, we find that XRCC4-XLF complexes robustly bridge two independent DNA molecules and that these bridges are able to slide along the DNA. These observations suggest that XRCC4-XLF complexes form mobile sleeve-like structures around DNA that can reconnect the broken ends very rapidly and hold them together. Understanding the dynamics and regulation of this mechanism will lead to clarification of how NHEJ proteins are involved in generating chromosomal translocations.


Reciprocal enhancement of gene expression and viral replication between human cytomegalovirus and human immunodeficiency virus type 1.
Biological interactions between human cytomegalovirus (HCMV) and the human immunodeficiency virus type 1 (HIV-1) were analysed in transfection and infection experiments, carried out in a human osteogenic sarcoma cell line (HOS) and in the same cell line chronically infected with HCMV (E155). When HOS and E155 cells were transfected with recombinant plasmids containing the HIV long terminal repeat (LTR) linked to the bacterial chloramphenicol acetyltransferase (CAT) gene, LTR-directed CAT expression was 20 times higher in E155 cells than in HOS cells. HOS cells co-infected with HCMV and HIV-1 showed enhanced production of the HIV-1 p24 antigen. In reciprocal experiments, an increase in HCMV immediate early gene expression was observed when HCMV-infected HOS cells and E155 cells were either transfected with a recombinant plasmid containing the HIV transactivator gene (pTAT), or when infected with HIV-1. DNA hybridization analysis of E155 and HCMV-infected HOS cells revealed higher levels of HCMV DNA in cells transfected with pTAT than in cells transfected with other non-specific recombinant plasmids. E155 cells transfected with pTAT also produced higher titres of infectious HCMV than control cultures of E155 cells transfected with other recombinant plasmids, including pMTAT carrying a mutant tat gene. The functional reciprocity in vitro between HCMV and HIV is discussed with respect to its possible implications for the clinical development of AIDS.


Comparing immunochromatography with latex antigen agglutination testing for the diagnosis of cryptococcosis in cats, dogs and koalas.
Although the point-of-care cryptococcal antigen lateral flow assay (LFA) has revolutionized the diagnosis of cryptococcosis in human patients, to date there has been no large-scale examination of this test in animals. We therefore assessed the LFA in cats, dogs and koalas suspected of having cryptococcosis. In sum, 528 serum specimens (129 from cats, 108 from dogs, 291 from koalas) were tested using the LFA and one of two commercially available latex cryptococcal antigen agglutination test (LCAT) kits. The LCAT is a proven and well-accepted method in veterinary patients and therefore taken as the "gold standard" against which the LFA was compared. The LFA achieved a sensitivity of 92%, 100%, and 98% in cats, dogs, and koalas, respectively, with corresponding negative predictive values of 94%, 100%, and 98%. The specificity of the LFA was 81%, 84%, and 62% for cats, dogs, and koalas, respectively, with corresponding positive predictive values of 76%, 48%, and 69%. These findings suggest the most appropriate role for the LFA is as a screening test to rule out a diagnosis of cryptococcosis in cats, dogs, and koalas. Point-of-care accessibility makes it equally suited for use in the field and as a cage-side test in veterinary hospitals. The suboptimal specificity of the LFA makes it less suited to definitive confirmation of cryptococcosis in animals; therefore, all LFA-positive test results should be confirmed by LCAT testing. The discrepancy between these observations and the high specificity of the LFA in humans may reflect differences in the host-pathogen interactions amongst the species.


Analysis of Risk Factors of Pancreatic Injury during Elective Laparoscopic Splenectomy in Children.
Laparoscopic splenectomy (LS) became the standard choice for splenectomy in children with benign hematological disease. There are few reports about pancreatic injury during LS. The purpose of this study is to spot on factors increasing the risk of pancreatic injury during LS in children. A total of 140 children had LS for benign causes. Children were categorized into A and B groups. LigaSure™ was used to control pedicle in Group A, while endoscopic staplers were used in Group B. Preoperative levels of amylase, lipase, and lactate dehydrogenase (LDH) were obtained. The mean of pancreatic enzymes and LDH values was calculated on the 3 postoperative successive days. A total of 71 boys and 69 girls had LS. The mean splenic size was 13.50 cm in Group A and 12.51 cm in Group B. The mean operative time in Group A was 41.91 min and in Group B was 56.36 min. The mean level of amylase was 42.99 IU/ml in Group A and 75.70 IU/ml in Group B (P = 0.001). The mean level of lipase was 37 IU/ml in Group A and 76.66 IU/ml in Group B (P = 0.001). Pancreatic injury during LS is a rare complication usually presented on biochemical level. We believe that it is a hemostatic-dependent complication rather than splenic size or nature of disease.


What is new for resveratrol? Is a new set of recommendations necessary?
Numerous scientific papers have suggested health-promoting effects of resveratrol, including claims in the prevention of diseases such as coronary heart disease, diabetes, and cancer. Therefore, it was proposed that the scientific community needed to express recommendations on the human use of resveratrol. Such recommendations were formulated after the first international resveratrol conference in Denmark, Resveratrol2010. The working group stated that the evidence was "not sufficiently strong to justify recommendation for the chronic administration of resveratrol to human beings, beyond the dose which can be obtained from dietary sources." It was a disappointing conclusion relative to the positive claims about the therapeutic potential of resveratrol made by the media. However, since 2010, results from the first clinical trials on resveratrol have been made available. Because of these emerging results, it is necessary to formulate updated versions of the recommendations.


Limited English proficiency and psychological distress among Latinos and Asian Americans.
English proficiency is increasingly recognized as an important factor that is related to the mental health of immigrants and ethnic minorities. However, few studies have examined how the association between English proficiency and mental health operates and whether the pattern of association is similar or different among various ethnic minority groups. This paper investigates how limited English proficiency directly and indirectly affects psychological distress through pathways of discrimination for both Latinos and Asian Americans in the United States. Findings suggest that, for Asian Americans, limited English proficiency has an independent relationship with psychological distress over and above demographic variables, socioeconomic and immigration-related factors and discrimination. For Latinos, however, socio-demographic variables and discrimination show a stronger association than limited English proficiency in affecting psychological distress. Different forms of discrimination - everyday discrimination and racial/ethnic discrimination - are equally important for both ethnic groups. Findings underscore the differential role of limited English proficiency for the mental health of Asian Americans and Latinos and suggest the distinctive racial experiences and backgrounds of these two ethnic groups.


Trophic effects of adipose-tissue-derived and bone-marrow-derived mesenchymal stem cells enhance cartilage generation by chondrocytes in co-culture.
Combining mesenchymal stem cells (MSCs) and chondrocytes has great potential for cell-based cartilage repair. However, there is much debate regarding the mechanisms behind this concept. We aimed to clarify the mechanisms that lead to chondrogenesis (chondrocyte driven MSC-differentiation versus MSC driven chondroinduction) and whether their effect was dependent on MSC-origin. Therefore, chondrogenesis of human adipose-tissue-derived MSCs (hAMSCs) and bone-marrow-derived MSCs (hBMSCs) combined with bovine articular chondrocytes (bACs) was compared. hAMSCs or hBMSCs were combined with bACs in alginate and cultured in vitro or implanted subcutaneously in mice. Cartilage formation was evaluated with biochemical, histological and biomechanical analyses. To further investigate the interactions between bACs and hMSCs, (1) co-culture, (2) pellet, (3) Transwell® and (4) conditioned media studies were conducted. The presence of hMSCs-either hAMSCs or hBMSCs-increased chondrogenesis in culture; deposition of GAG was most evidently enhanced in hBMSC/bACs. This effect was similar when hMSCs and bAC were combined in pellet culture, in alginate culture or when conditioned media of hMSCs were used on bAC. Species-specific gene-expression analyses demonstrated that aggrecan was expressed by bACs only, indicating a predominantly trophic role for hMSCs. Collagen-10-gene expression of bACs was not affected by hBMSCs, but slightly enhanced by hAMSCs. After in-vivo implantation, hAMSC/bACs and hBMSC/bACs had similar cartilage matrix production, both appeared stable and did not calcify. This study demonstrates that replacing 80% of bACs by either hAMSCs or hBMSCs does not influence cartilage matrix production or stability. The remaining chondrocytes produce more matrix due to trophic factors produced by hMSCs.


Clinical care for opioid-using pregnant and postpartum women: the role of obstetric providers.
We review clinical care issues that are related to illicit and therapeutic opioid use among pregnant women and women in the postpartum period and outline the major responsibilities of obstetrics providers who care for these patients during the antepartum, intrapartum, and postpartum periods. Selected patient treatment issues are highlighted, and case examples are provided. Securing a strong rapport and trust with these patients is crucial for success in delivering high-quality obstetric care and in coordinating services with other specialists as needed. Obstetrics providers have an ethical obligation to screen, assess, and provide brief interventions and referral to specialized treatment for patients with drug use disorders. Opioid-dependent pregnant women often can be treated effectively with methadone or buprenorphine. These medications are classified as pregnancy category C medications by the Food and Drug Administration, and their use in the treatment of opioid-dependent pregnant patients should not be considered "off-label." Except in rare special circumstances, medication-assisted withdrawal during pregnancy should be discouraged because of a high relapse rate. Acute pain management in this population deserves special consideration because patients who use opioids can be hypersensitive to pain and because the use of mixed opioid-agonist/antagonists can precipitate opioid withdrawal. In the absence of other indications, pregnant women who use opioids do not require more intense medical care than other pregnant patients to ensure adequate treatment and the best possible outcomes. Together with specialists in pain and addiction medicine, obstetricians can coordinate comprehensive care for pregnant women who use opioids and women who use opioids in the postpartum period.


Physical therapist-delivered cognitive-behavioral therapy: a qualitative study of physical therapists' perceptions and experiences.
The importance of the biopsychosocial model in assessment and management of chronic musculoskeletal conditions is recognized. Physical therapists have been encouraged to develop psychologically informed practice. Little is known about the process of physical therapists' learning and delivering of psychological interventions within the practice context. The aim of this study was to investigate physical therapists' experiences and perspectives of a cognitive-behavioral-informed training and intervention process as part of a randomized controlled trial (RCT) involving adults with painful knee osteoarthritis. A qualitative design was used. Participants were physical therapists trained to deliver pain coping skills training (PCST). Eight physical therapists trained to deliver PCST were interviewed by telephone at 4 time points during the 12-month RCT period. Interviews were audiorecorded, transcribed verbatim into computer-readable files, and analyzed using Framework Analysis. Thematic categories identified were: training, experience delivering PCST, impact on general clinical practice, and perspectives on PCST and physical therapist practice. Physical therapists reported positive experiences with PCST and program delivery. They thought that their participation in the RCT had enhanced their general practice. Although some components of the PCST program were familiar, the therapists found delivering the program was quite different from regular practice. Physical therapists believed the PCST program, a 3- to 4-day workshop followed by formal mentoring and performance feedback from a psychologist for 3 to 6 months and during the RCT, was critical to their ability to effectively deliver the PCST intervention. They identified a number of challenges in delivering PCST in their normal practice. Physical therapists can be trained to confidently deliver a PCST program. The physical therapists in this study believed that training enhanced their clinical practice. Comprehensive training and mentoring by psychologists was crucial to ensure treatment fidelity.


Comparison of seroprevalence of SARS-CoV-2 infections with cumulative and imputed COVID-19 cases: Systematic review.
Accurate seroprevalence estimates of SARS-CoV-2 in different populations could clarify the extent to which current testing strategies are identifying all active infection, and hence the true magnitude and spread of the infection. Our primary objective was to identify valid seroprevalence studies of SARS-CoV-2 infection and compare their estimates with the reported, and imputed, COVID-19 case rates within the same population at the same time point. We searched PubMed, Embase, the Cochrane COVID-19 trials, and Europe-PMC for published studies and pre-prints that reported anti-SARS-CoV-2 IgG, IgM and/or IgA antibodies for serosurveys of the general community from 1 Jan to 12 Aug 2020. Of the 2199 studies identified, 170 were assessed for full text and 17 studies representing 15 regions and 118,297 subjects were includable. The seroprevalence proportions in 8 studies ranged between 1%-10%, with 5 studies under 1%, and 4 over 10%-from the notably hard-hit regions of Gangelt, Germany; Northwest Iran; Buenos Aires, Argentina; and Stockholm, Sweden. For seropositive cases who were not previously identified as COVID-19 cases, the majority had prior COVID-like symptoms. The estimated seroprevalences ranged from 0.56-717 times greater than the number of reported cumulative cases-half of the studies reported greater than 10 times more SARS-CoV-2 infections than the cumulative number of cases. The findings show SARS-CoV-2 seroprevalence is well below "herd immunity" in all countries studied. The estimated number of infections, however, were much greater than the number of reported cases and deaths in almost all locations. The majority of seropositive people reported prior COVID-like symptoms, suggesting that undertesting of symptomatic people may be causing a substantial under-ascertainment of SARS-CoV-2 infections.


Experimental demyelinating optic neuropathy: a model for combined morphological and electrophysiological studies.
A model is described for the morphological and electrophysiological study of demyelinative optic neuropathy in the rat. Cortical visual evoked potentials (VEPs) to flash stimuli were recorded before and after the intraneural microinjection of lysophosphatidyl choline (LPC) and galacto-cerebroside (Gal-C) antiserum. Progress was monitored by calculating the interocular latency differences (ILD) for the N40 and P60 components of the rat VEP, thus allowing a longitudinal evaluation of optic nerve function. Histological examination of the visual pathway using light and electron microscopy was performed at intervals through the course of the study. LPC produced a less selective lesion than did Gal-C antiserum. In lesions induced by the latter there was relatively little accompanying anoxal degeneration and subsequent remyelination was prominent. Control serum did not result in demyelination. This model should permit accurate electrophysiological and morphological correlations to be made during the developing and the reparative phases of demyelinative optic neuropathy and the assessment of potential therapeutic manoeuvres.


Prevention of serious respiratory syncytial virus-related illness. II: Immunoprophylaxis.
Respiratory syncytial virus (RSV) causes significant morbidity in very young children, preterm infants with and without chronic lung disease, and children with hemodynamically significant congenital heart disease. In the absence of a safe and effective vaccine, alternative means of protecting high-risk infants and young children from serious RSV illness have been studied. Clinical observations and animal model data over the past 30 years suggested that RSV immunoglobulin G (IgG) neutralizing antibodies might offer protection from severe RSV lower respiratory tract disease. Transfer of adequate amounts of IgG to the fetus does not occur efficiently until the third trimester of pregnancy, which helps to explain why premature infants are at high risk of serious RSV illness. Efforts shifted toward the prophylactic monthly administration of standard immunoglobulins and, later, of RSV-enriched immunoglobulin in selected high-risk infants and young children. Although this approach proved effective, RSV-enriched immune globulin was not suitable for all patients and administration was labor intensive. The development of palivizumab, a monoclonal antibody that can bind to a specific antigenic site on the virus and prevent cell-to-cell spread of infection has since become the mainstay of RSV illness prevention in preterm infants and those with significant congenital heart disease. Palivizumab, the only monoclonal antibody approved for the prevention of RSV lower respiratory tract disease must be administered monthly throughout the RSV season and does not always prevent serious RSV illness. Further research to develop more effective and less labor-intensive immunoprophylactic agents is ongoing.


Automatic Estimation of Coronary Blood Flow Velocity Step 1 for Developing a Tool to Diagnose Patients With Micro-Vascular Angina Pectoris.
Aim: Our aim was to automatically estimate the blood velocity in coronary arteries using cine X-ray angiographic sequence. Estimating the coronary blood velocity is a key approach in investigating patients with angina pectoris and no significant coronary artery disease. Blood velocity estimation is central in assessing coronary flow reserve. Methods and Results: A multi-step automatic method for blood flow velocity estimation based on the information extracted solely from the cine X-ray coronary angiography sequence obtained by invasive selective coronary catheterization was developed. The method includes (1) an iterative process of segmenting coronary arteries modeling and removing the heart motion using a non-rigid registration, (2) measuring the area of the segmented arteries in each frame, (3) fitting the measured sequence of areas with a 7° polynomial to find start and stop time of dye propagation, and (4) estimating the blood flow velocity based on the time of the dye propagation and the length of the artery-tree. To evaluate the method, coronary angiography recordings from 21 patients with no obstructive coronary artery disease were used. In addition, coronary flow velocity was measured in the same patients using a modified transthoracic Doppler assessment of the left anterior descending artery. We found a moderate but statistically significant correlation between flow velocity assessed by trans thoracic Doppler and the proposed method applying both Spearman and Pearson tests. Conclusion: Measures of coronary flow velocity using a novel fully automatic method that utilizes the information from the X-ray coronary angiographic sequence were statistically significantly correlated to measurements obtained with transthoracic Doppler recordings.


Compact Fourier-transform volume holographic spectrometer for diffuse source spectroscopy.
We present a new idea for diffuse source spectroscopy using a Fourier-transform volume holographic spectrometer formed by a Fourier-transform lens, a volume hologram, and a CCD. We show that this spectrometer can operate well under spatially incoherent light illumination. Furthermore, this spectrometer is less bulky, less sensitive to input alignment, and potentially more appropriate for implementation of highly sensitive spectrometers than conventional spectrometers.


Biplane wing planform and flight performance of the feathered dinosaur Microraptor gui.
Microraptor gui, a four-winged dromaeosaur from the Early Cretaceous of China, provides strong evidence for an arboreal-gliding origin of avian flight. It possessed asymmetric flight feathers not only on the manus but also on the pes. A previously published reconstruction shows that the hindwing of Microraptor supported by a laterally extended leg would have formed a second pair of wings in tetrapteryx fashion. However, this wing design conflicts with known theropod limb joints that entail a parasagittal posture of the hindlimb. Here, we offer an alternative planform of the hindwing of Microraptor that is concordant with its feather orientation for producing lift and normal theropod hindlimb posture. In this reconstruction, the wings of Microraptor could have resembled a staggered biplane configuration during flight, where the forewing formed the dorsal wing and the metatarsal wing formed the ventral one. The contour feathers on the tibia were positioned posteriorly, oriented in a vertical plane for streamlining that would reduce the drag considerably. Leg feathers are present in many fossil dromaeosaurs, early birds, and living raptors, and they play an important role in flight during catching and carrying prey. A computer simulation of the flight performance of Microraptor suggests that its biplane wings were adapted for undulatory "phugoid" gliding between trees, where the horizontal feathered tail offered additional lift and stability and controlled pitch. Like the Wright 1903 Flyer, Microraptor, a gliding relative of early birds, took to the air with two sets of wings.


Evaluating Molecular Biomarkers for the Early Detection of Lung Cancer: When Is a Biomarker Ready for Clinical Use? An Official American Thoracic Society Policy Statement.
Molecular biomarkers have the potential to improve the current state of early lung cancer detection. The goal of this project was to develop a policy statement that provides guidance about the level of evidence required to determine that a molecular biomarker, used to support early lung cancer detection, is appropriate for clinical use. An ad hoc project steering committee was formed, to include individuals with expertise in the early detection of lung cancer and molecular biomarker development, from inside and outside of the Assembly on Thoracic Oncology. Key questions, generated from the results of a survey of the project steering committee, were discussed at an in-person meeting. Results of the discussion were summarized in a policy statement that was circulated to the steering committee and revised multiple times to achieve consensus. With a focus on the clinical applications of lung cancer screening and lung nodule evaluation, the policy statement outlines categories of results that should be reported in the early phases of molecular biomarker development, discusses the level of evidence that would support study of the clinical utility, describes the outcomes that should be proven to consider a molecular biomarker clinically useful, and suggests study designs capable of assessing these outcomes. The application of molecular biomarkers to assist with the early detection of lung cancer has the potential to substantially improve our ability to select patients for lung cancer screening, and to assist with the characterization of indeterminate lung nodules. We have described relevant considerations and have suggested standards to apply when determining whether a molecular biomarker for the early detection of lung cancer is ready for clinical use.


Understanding the unique experiences, perspectives and sexual and reproductive health needs of very young adolescents: Somali refugees in Ethiopia.
Kobe Refugee camp hosts roughly 39,000 refugees displaced from Somalia during the 2011-2012 Horn of Africa Crisis. Sexual and reproductive health, as with the greater issues of health and well-being for adolescents displaced from this crisis remain largely unknown and neglected. In 2013, the Women's Refugee Commission, Johns Hopkins University, and International Medical Corps in Ethiopia, implemented qualitative and quantitative research to explore the factors and risks that impact the health of very young adolescents (VYAs), those 10-14 years of age, in this setting. This paper presents findings from the qualitative effort. Focus group discussions (FGD), incorporating community mapping and photo elicitation activities, were conducted with 10-12 and 13-14 year-olds to obtain information about their own perspectives, experiences and values. FGDs were also implemented with 15-16 year-olds and adults, to consider their perspectives on the sexual and reproductive health needs and risks of VYAs. This research identified several factors that were found to influence the health and well-being of VYAs in Kobe refugee camp, including newfound access to education and security, combined with gender divisions and parental communication around early SRH and puberty that remained intact from traditional Somali culture. Girls were found to face an additional risk of child marriage and early pregnancy exacerbated since displacement, which significantly limited their ability to access education and achieve future aspirations. Findings from this study could help to inform future programs in Kobe and similar contexts involving long-term displacement from conflict, focusing on the health and development needs of VYAs. Future programs should consider the determinants of positive VYA health and development, including access to education, gender equity, and safety.By better understanding the unique experiences, perspectives and needs of VYAs, practitioners, policy makers and donors can invest in the individual and community assets that reinforce positive behaviors established in early adolescence, in order to achieve long-term SRH impacts.


The Personal Genome Project Canada: findings from whole genome sequences of the inaugural 56 participants.
The Personal Genome Project Canada is a comprehensive public data resource that integrates whole genome sequencing data and health information. We describe genomic variation identified in the initial recruitment cohort of 56 volunteers. Volunteers were screened for eligibility and provided informed consent for open data sharing. Using blood DNA, we performed whole genome sequencing and identified all possible classes of DNA variants. A genetic counsellor explained the implication of the results to each participant. Whole genome sequencing of the first 56 participants identified 207 662 805 sequence variants and 27 494 copy number variations. We analyzed a prioritized disease-associated data set (n = 1606 variants) according to standardized guidelines, and interpreted 19 variants in 14 participants (25%) as having obvious health implications. Six of these variants (e.g., in BRCA1 or mosaic loss of an X chromosome) were pathogenic or likely pathogenic. Seven were risk factors for cancer, cardiovascular or neurobehavioural conditions. Four other variants - associated with cancer, cardiac or neurodegenerative phenotypes - remained of uncertain significance because of discrepancies among databases. We also identified a large structural chromosome aberration and a likely pathogenic mitochondrial variant. There were 172 recessive disease alleles (e.g., 5 individuals carried mutations for cystic fibrosis). Pharmacogenomics analyses revealed another 3.9 potentially relevant genotypes per individual. Our analyses identified a spectrum of genetic variants with potential health impact in 25% of participants. When also considering recessive alleles and variants with potential pharmacologic relevance, all 56 participants had medically relevant findings. Although access is mostly limited to research, whole genome sequencing can provide specific and novel information with the potential of major impact for health care.


Aqueous two-phase microdroplets with reversible phase transitions.
Aqueous two-phase systems contained within microdroplets enable a bottom-up approach to mimicking the dynamic microcompartmentation of biomaterial that naturally occurs within the cytoplasm of cells. Here, we demonstrate the generation of femtolitre aqueous two-phase droplets within a microfluidic oil channel. Gated pressure pulses were used to generate individual, stationary two-phase microdroplets with a well-defined time zero for carrying out controlled and sequential phase transformations over time. Reversible phase transitions between single-phase, two-phase, and core-shell microbead states were obtained via evaporation-induced dehydration and water rehydration. In contrast to other microfluidic aqueous two-phase droplets, which require continuous flows and high-frequency droplet formation, our system enables the controlled isolation and reversible transformation of a single microdroplet and is expected to be useful for future studies in dynamic microcompartmentation and affinity partitioning.


Accelerated evolution of 3'avian FOXE1 genes, and thyroid and feather specific expression of chicken FoxE1.
The forkhead transcription factor gene E1 (FOXE1) plays an important role in regulation of thyroid development, palate formation and hair morphogenesis in mammals. However, avian FOXE1 genes have not been characterized and as such, codon evolution of FOXE1 orthologs in a broader evolutionary context of mammals and birds is not known. In this study we identified the avian FOXE1 gene in chicken, turkey and zebra finch, all of which consist of a single exon. Chicken and zebra finch FOXE1 are uniquely located on the sex-determining Z chromosome. In situ hybridization shows that chicken FOXE1 is specifically expressed in the developing thyroid. Its expression is initiated at the placode stage and is maintained during the stages of vesicle formation and follicle primordia. Based on this expression pattern, we propose that avian FOXE1 may be involved in regulating the evagination and morphogenesis of thyroid. Chicken FOXE1 is also expressed in growing feathers. Sequence analysis identified two microdeletions in the avian FOXE1 genes, corresponding to the loss of a transferable repression domain and an engrailed homology motif 1 (Eh1) C-terminal to the forkhead domain. The avian FOXE1 proteins exhibit a significant sequence divergence of the C-terminus compared to those of amphibian and mammalian FOXE1. The codon evolution analysis (dN/dS) of FOXE1 shows a significantly increased dN/dS ratio in the avian lineages, consistent with either a relaxed purifying selection or positive selection on a few residues in avian FOXE1 evolution. Further site specific analysis indicates that while relaxed purifying selection is likely to be a predominant cause of accelerated evolution at the 3'-region of avian FOXE1, a few residues might have evolved under positive selection. We have identified three avian FOXE1 genes based on synteny and sequence similarity as well as characterized the expression pattern of the chicken FOXE1 gene during development. Our evolutionary analyses suggest that while a relaxed purifying selection is likely to be the dominant force driving accelerated evolution of avian FOXE1 genes, a few residues may have evolved adaptively. This study provides a basis for future genetic and comparative biochemical studies of FOXE1.


Evidences of hepatoprotective and antioxidant effect of Citrullus colocynthis fruits in paracetamol induced hepatotoxicity.
The objective of present study was to explore the hepatoprotective and antioxidant profile of Citrullus colocynthis fruits. Hepatoprotective profile of methanolic extract of Citrullus colocynthis fruits (MECCF) was investigated on rats, which were made hepatotoxic using paracetamol. The antioxidant profile of MECCF was evaluated by conducting Catalase, Super oxide Dismutase, Lipid Peroxidation and Diphenyl Picryl Hydrazyl tests. During hepatoprotective investigation, the Paracetamol treated group II showed significant increase in total bilirubin (TB), serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) and alkaline phosphatase (ALP) level. The results so obtained showed that pretreatment of rats with MECCF 300mg/kg p.o. decreases the elevated TB, SGOT, SGPT and ALP serum levels. Also, MECCF inhibitory profile was found comparable with toxicant group (Paracetamol 2g/kg, p.o.). The present study concludes that MECCF fruit possess significant hepatoprotective and antioxidant activity.


Quantification of risk from fetal exposure to diagnostic ultrasound.
Biomedical ultrasound may induce adverse effects in patients by either thermal or non-thermal means. Temperatures above normal can adversely affect biological systems, but effects also may be produced without significant heating. Thermally induced teratogenesis has been demonstrated in many animal species as well as in a few controlled studies in humans. Various maximum 'safe' temperature elevations have been proposed, although the suggested values range from 0.0 to 2.5 degrees C. Factors relevant to thermal effects are considered, including the nature of the acoustic field in situ, the state of perfusion of the embryo/fetus, and the variation of sensitivity to thermal insult with gestational stage of development. Non-thermal mechanisms of action considered include acoustic cavitation, radiation force, and acoustic streaming. While cavitation can be quite destructive, it is extremely unlikely in the absence of stabilized gas bodies, and although the remaining mechanisms may occur in utero, they have not been shown to induce adverse effects. For example, pulsed, diagnostic ultrasound can increase fetal activity during exposure, apparently due to stimulation of auditory perception by radiation forces on the fetal head or auditory structures. In contrast, pulsed ultrasound also produces vascular damage near developing bone in the late-gestation mouse, but by a unknown mechanism and at levels above current US FDA output limits. It is concluded that: (1) thermal rather than nonthermal mechanisms are more likely to induce adverse effects in utero, and (2) while the probability of an adverse thermal event is usually small, under some conditions it can be disturbingly high.