Case Report: Atraumatic Orbital Emphysema after Bronchoscopy with Endobronchial Biopsy.
Orbital emphysema is a well-documented condition where air enters the orbital soft tissue typically because of direct or indirect trauma; however, atraumatic cases can be encountered. The following is a novel case report of acute unilateral atraumatic orbital emphysema 24 hours after bronchoscopy with endobronchial biopsy. To the best of the authors' knowledge, this is the first known case report of atraumatic orbital emphysema as a postprocedure complication of bronchoscopy with endobronchial biopsy in the literature. A 74-year-old man presented with painless swelling of the left upper and lower eyelids upon awakening. The patient had a history of bronchoscopy and endobronchial biopsy with conscious sedation for a right upper lobe lung mass 24 hours before the visit. Slit-lamp examination revealed air entrapment in the left upper and lower lids with enlarged subconjunctival air pockets of the left eye. Head and maxillofacial computed tomography without contrast revealed extensive emphysema tracking and dissecting along the subcutaneous and deep soft tissue spaces of the neck and face that was more pronounced along the left side. The patient was admitted for overnight observation and prescribed a prophylactic oral course of 250 mg of azithromycin daily for 2 days by the attending hospitalist. The subsequent follow-up visit 3 days later revealed complete resolution of signs and symptoms. This case report demonstrates the importance of considering orbital emphysema in patients with a history of recent bronchoscopy with endobronchial biopsy. Clinicians should be aware of this potential complication and refer for appropriate testing and comanagement.


A Modified Broström Repair with Transosseous Fixation for Chronic Ankle Instability: A Midterm Followup Study in Soldiers.
Various surgical techniques are available to reduce chronic instability of the lateral ankle ligament complex. The most effective method for these procedures remains controversial. This report presents a surgical technique that is similar to the Broström procedure and uses a modified, nonaugmented repair technique. 38 soldiers with a history of chronic lateral ankle instability and poor ankle function underwent plication of the anterior talofibular ligament-lateral capsule complex with transosseous fixation of the calcaneofibular ligament through a fibular bone tunnel between 2004 and 2007. This study included 33 men and 5 women with a mean age of 25.6 years (range 18-36 years) at the time of surgery. Each patient was confirmed to have a history of chronic lateral ankle instability after an inversion injury, and symptoms had been noted for at least 1 year. The patients were followed up with stress radiographs, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot functional score, and the Sefton assessment system. The mean followup period was 77.6 months (range 66-89 months). At the last evaluation, the talar tilt reduced from an average of 13.9° ± 2.4° before surgery to 3.8° ± 1.8° after surgery, and anterior drawer displacement reduced from 9.6 ± 2.9 mm to 2.3 ± 1.6 mm. The mean AOFAS ankle-hindfoot scale score for functional stability increased from 71.6 ± 4.0 points preoperatively to 95.6 ± 4.0 points postoperatively. As evaluated by the Sefton assessment system, 36 patients (95%) reported an excellent or good functional outcome. All patients resumed normal daily activities and active military duty after the surgery. The procedure described here could be considered a viable alternative option to anatomic reconstruction such as the modified Broström procedure and might be appropriate for the general population.


Tuberculosis is the leading cause of lymphadenopathy in HIV-infected persons in India: results of a fine-needle aspiration analysis.
HIV infection is associated with a number of opportunistic infections and malignancies frequently involving the lymph nodes. Lymphadenopathy may occur at any stage of HIV infection. We aimed to determine the utility of fine-needle aspiration cytology in evaluating the causes of lymphadenopathy in HIV-infected individuals. Three hundred HIV-infected individuals with lymphadenopathy were included in the study. Fine-needle aspiration (FNA) was performed on peripheral or deep-seated lymph nodes. The material was used for cytological examination using May-Grunwald-Giemsa and haematoxylin and eosin staining. Special stains such as modified Ziehl-Neelsen staining for acid-fast bacilli and periodic acid-Schiff staining for fungi were also performed. The mean age of the study group was 35.0 ± 8.0 y (range 13-74 y). The median CD4 count was 152 cells/μl. Out of the 300 FNA reports, acid-fast bacteria were reported in 130 and cytological findings indicating mycobacterial infection in a further 43 patients. Cryptococcosis was reported in 4 individuals, histoplasmosis in 2 and aspergillosis in 1. Reactive hyperplasia was seen in 89 individuals. Lymphoma was noted in 7 individuals and suppurative inflammation in 5. In conclusion, tuberculosis is the predominant cause of lymphadenitis in HIV-infected individuals in India, especially in those with low CD4 cell counts.


Alcohol Accessibility and Family Violence-related Ambulance Attendances.
There is substantial evidence supporting the association between alcohol license density and violent crime. However, the impact of different types of alcohol licenses on intimate partner and family violence is sparse. We explored the associations between access to alcohol outlets, and family and intimate partner violence using paramedic clinical records, given this service is often the first to respond to acute crises. Coded ambulance attendance data from 694 postcodes in Victoria, Australia, from July 1, 2016 to June 30, 2018 where alcohol or another drug, mental health or self-harm associated with family or intimate partner violence was indicated were examined. A hybrid model of spatial autoregressive and negative binomial zero-inflated Poisson-based count regression models was used to examine associations with alcohol outlet density and socioeconomic factors. We found that access to a liquor license outlet was significantly associated with family violence-related attendances across all types of outlets, including on-premise (late night) licenses (β = 1.73, SE: 0.18), restaurant licenses (β = 0.83, SE: 0.28), and packaged liquor licenses (β = 0.62, SE: 0.06). Our results demonstrate a significant relationship between alcohol-related harms in the context of family violence and provides evidence of the relationship between alcohol-related family violence in both victims and perpetrators. The findings of this study highlight the need for public health interventions such as licensing policy and town planning changes to reduce these harms by restricting alcohol availability.


Phosphate Transporter PstSCAB of Campylobacter jejuni Is a Critical Determinant of Lactate-Dependent Growth and Colonization in Chickens.
Campylobacter jejuni causes acute gastroenteritis worldwide and is transmitted primarily through poultry, in which it is often a commensal member of the intestinal microbiota. Previous transcriptome sequencing (RNA-Seq) experiment showed that transcripts from an operon encoding a high-affinity phosphate transporter (PstSCAB) of C. jejuni were among the most abundant when the bacterium was grown in chickens. Elevated levels of the pstSCAB mRNA were also identified in an RNA-Seq experiment from human infection studies. In this study, we explore the role of PstSCAB in the biology and colonization potential of C. jejuni Our results demonstrate that cells lacking PstSCAB survive poorly in stationary phase, in nutrient-limiting media, and under osmotic conditions reflective of those in the chicken. Polyphosphate levels in the mutant cells were elevated at stationary phase, consistent with alterations in expression of polyphosphate metabolism genes. The mutant strain was highly attenuated for colonization of newly hatched chicks, with levels of bacteria at several orders of magnitude below wild-type levels. Mutant and wild type grew similarly in complex media, but the pstS::kan mutant exhibited a significant growth defect in minimal medium supplemented with l-lactate, postulated as a carbon source in vivo Poor growth in lactate correlated with diminished expression of acetogenesis pathway genes previously demonstrated as important for colonizing chickens. The phosphate transport system is thus essential for diverse aspects of C. jejuni physiology and in vivo fitness and survival.IMPORTANCECampylobacter jejuni causes millions of human gastrointestinal infections annually, with poultry a major source of infection. Due to the emergence of multidrug resistance in C. jejuni, there is need to identify alternative ways to control this pathogen. Genes encoding the high-affinity phosphate transporter PstSCAB are highly expressed by C. jejuni in chickens and humans. In this study, we address the role of PstSCAB on chicken colonization and other C. jejuni phenotypes. PstSCAB is required for colonization in chicken, metabolism and survival under different stress responses, and during growth on lactate, a potential growth substrate in chickens. Our study highlights that PstSCAB may be an effective target to develop mechanisms for controlling bacterial burden in both chicken and human.


Topical delta 9-tetrahydrocannabinol and aqueous dynamics in glaucoma.
Systemic delta 9-tetrahydrocannabinol (THC), administered either by smoking marihuana or as synthetic THC in soft gelatin capsules, lowers ocular tension in various glaucomas, but at the expense of significant decreases in systolic blood pressure. Topical THC in light mineral oil vehicles, though effective in laboratory animals, was not shown effective in 0.05 and 0.1% topical solutions when administered to six subjects with primary open-angle glaucoma in a randomized, balanced, double-masked protocol. Light mineral oil, which has an affinity for corneal epithelium, is an optimum vehicle for administering drugs whose mechanisms of action are systemic rather than local within the eye. Further glaucoma research should therefore proceed with marihuanas containing insignificant levels of THC (less than 0.4%) and with various local delivery systems of the ocular-active cannabinoid found in Cannabis sativa.


Impaired working-memory after cerebellar infarcts paralleled by changes in BOLD signal of a cortico-cerebellar circuit.
A considerable body of evidence supports the notion that cerebellar lesions lead to neuropsychological deficits, including impairments in working-memory, executive tasks and verbal fluency. Studies employing functional magnetic resonance imaging (fMRI) and anatomical tracing in primates provide evidence for a cortico-cerebellar circuitry as the functional substrate of working-memory. The present fMRI study explores the activation pattern during an n-back working-memory task in patients with an isolated cerebellar infarct. To determine each patient's cognitive impairment, neuropsychological tests of working-memory and attention were carried out. We conducted fMRI in nine patients and nine healthy age-matched controls while they performed a 2-back task in a blocked-design. In both groups we found bilateral activations in a widespread cortico-cerebellar network, consisting of the ventrolateral prefrontal cortex (BA 44, 45), dorsolateral prefrontal cortex (BA 9, 46), parietal cortex (BA 7, 40), pre-supplementary motor area (BA 6) anterior cingulate (BA 32). Relative to healthy controls, patients with isolated cerebellar infarcts demonstrated significantly more pronounced BOLD-activations in the precuneus and the angular gyrus during the 2-back task. The significant increase in activation in the posterior parietal areas of the cerebellar patients could be attributed to a compensatory recruitment to maintain task performance. We conclude that cerebellar lesions affect remote cortical regions that are part of a putative cortico-cerebellar network.


Common variants conferring risk of schizophrenia.
Schizophrenia is a complex disorder, caused by both genetic and environmental factors and their interactions. Research on pathogenesis has traditionally focused on neurotransmitter systems in the brain, particularly those involving dopamine. Schizophrenia has been considered a separate disease for over a century, but in the absence of clear biological markers, diagnosis has historically been based on signs and symptoms. A fundamental message emerging from genome-wide association studies of copy number variations (CNVs) associated with the disease is that its genetic basis does not necessarily conform to classical nosological disease boundaries. Certain CNVs confer not only high relative risk of schizophrenia but also of other psychiatric disorders. The structural variations associated with schizophrenia can involve several genes and the phenotypic syndromes, or the 'genomic disorders', have not yet been characterized. Single nucleotide polymorphism (SNP)-based genome-wide association studies with the potential to implicate individual genes in complex diseases may reveal underlying biological pathways. Here we combined SNP data from several large genome-wide scans and followed up the most significant association signals. We found significant association with several markers spanning the major histocompatibility complex (MHC) region on chromosome 6p21.3-22.1, a marker located upstream of the neurogranin gene (NRGN) on 11q24.2 and a marker in intron four of transcription factor 4 (TCF4) on 18q21.2. Our findings implicating the MHC region are consistent with an immune component to schizophrenia risk, whereas the association with NRGN and TCF4 points to perturbation of pathways involved in brain development, memory and cognition.


X-ray spectroscopic study of solvent effects on the ferrous and ferric hexacyanide anions.
We present an Fe Kα resonant inelastic X-ray scattering (RIXS) and X-ray emission (XES) study of ferrous and ferric hexacyanide dissolved in water and ethylene glycol. We observe that transitions below the absorption edge show that the solvent has a distinct effect on the valence electronic structure. In addition, both the RIXS and XES spectra show a stabilization of the 2p levels when dissolved in water. Using molecular dynamics simulations, we propose that this effect arises from the hydrogen-bonding interactions between the complex and nearby solvent molecules. This withdraws electron density from the ligands, stabilizing the complex but also causing a slight increase in π-backbonding.


Heterogeneity of the PorB protein in serotype 22 Neisseria meningitidis.
The genetic diversity of porB genes from meningococcal isolates characterized as serotype 22 was investigated by gene sequencing. This procedure identified seven distinct porB sequences, demonstrating variation in the PorB protein recognized by the serotype 22 monoclonal antibody. This is consistent with the genetic heterogeneity of serotype 22 meningococci reported previously.


Selectivity of bis(calix[4]diquinone) ionophores towards metal ions in solvent dimethylsulfoxide: a molecular mechanics and molecular dynamics study.
Molecular modelling studies have been carried out on two bis(calix[4]diquinone) ionophores, each created from two (calix[4]diquinone)arenes bridged at their bottom rims via alkyl chains (CH(2))(n), 1: n = 3, 2; n = 4, in order to understand the reported selectivity of these ligands towards different sized metal ions such as Na(+), K(+), Rb(+), and Cs(+) in dmso solution. Conformational analyses have been carried out which show that in the lowest energy conformations of the two macrocycles, the individual calix[4]diquinones exhibit a combination of partial cone, 1,3-alternate and cone conformations. The interactions of these alkali metals with the macrocycles have been studied in the gas phase and in a periodic box of solvent dmso by molecular mechanics and molecular dynamics calculations. Molecular mechanics calculations have been carried out on the mode of entry of the ions into the macrocycles and suggest that this is likely to occur from the side of the central cavity, rather than through the main axis of the calix[4]diquinones. There are energy barriers of ca. 19 kcal mol(-1) for this entry path in the gas phase, but in solution no energy barrier is found. Molecular dynamics simulations show that in both 1 and 2, though particularly in the latter macrocycle, one or two solvent molecules are bonded to the metal throughout the course of the simulation, often to the exclusion of one or more of the ether oxygen atoms. By contrast the carbonyl oxygen atoms remain bonded to the metal atoms throughout with bond lengths that remain significantly less than those to the ether oxygen atoms. Free energy perturbation studies have been carried out in dmso and indicate that for 1, the selectivity follows the order Rb(+) approximately K(+) > Cs(+) >> Na(+), which is partially in agreement with the experimental results. The energy differences are small and indeed the ratio between stability constants found for Cs(+) and K(+) complexes is only 0.60, showing that has only a slight preference for K(+). For the larger receptor , which is better suited to metal complexation, the binding affinity follows the pattern Cs(+) >> Rb(+) >> K(+) >> Na(+), with energy differences of 5.75, 2.61, 2.78 kcal mol(-1) which is perfectly consistent with experimental results.


Susceptibility changes following bolus injections.
The general mechanism of bulk magnetic susceptibility (BMS) induced MRI contrast following a bolus injection is elaborated. Combining radiolabeled tracer data for the first pass of a bolus injection through the human brain with the application of Wiedemann's law allows us to calculate the lower limit for the time course of the vascular BMS following the injection of any contrast agent. Superparamagnetic iron oxide particles produce a much larger effect than any mononuclear Ln(III) chelate. We also calculate the BMS changes occurring after a dilution bolus injection (of isosmolal physiological saline) subsequent to a prior slow infusion of an intravascular contrast agent. This technique bears some resemblance to the increasingly important approach that exploits changes in only the level of blood oxygenation. The calculation indicates that contrast changes after the dilution bolus injection are smaller than those following Ln(III) agent injections but larger than those due to changes in blood oxygenation and suggests a way to possibly enhance the latter. We present an in vivo study demonstrating the dilution bolus injection technique in the mouse brain, and that features its rapid repeatability. Extrapolation of these results to the human, however, indicates that the saline volumes required for venous injections, except possibly for cardiac studies, would be prohibitively large. Smaller, catheter-delivered arterial bolus injections are feasible. We also suggest a method for using an agent bolus injection to measure the parenchymal BMS, and thus the iron content, of pathologically iron-loaded tissue.


Sex differences in stroke recovery.
This study examined differences between men and women in the ability to perform basic activities of daily living, instrumental activities of daily living, and higher physical functioning after stroke. The objective of the study was to determine whether sex differences in stroke recovery can be explained by depressive status beyond older age, stroke severity, prestroke physical functioning, and other medical comorbidities. A total of 459 stroke patients were recruited from acute and subacute facilities in an urban midwestern community. These patients were followed prospectively from stroke onset until 6 months poststroke. All study participants were assessed using standardized stroke outcome measures, including the National Institutes of Health Stroke Scale, the Barthel Activities of Daily Living Index, the Lawton Instrumental Activities of Daily Living scale, and the SF-36 Health Survey physical functioning scale. The Geriatric Depression Scale was used to assess depressive status. Each outcome was measured at baseline (within 2 weeks of stroke onset), as well as 1, 3, and 6 months poststroke. Prestroke physical functioning, stroke characteristics, and comorbidities were also assessed at baseline. Female patients in the study were older than male patients, with a mean age of 71 years for women vs 69 years for men. Female patients reported lower prestroke physical functioning than their male counterparts. Six months after stroke, women in the study were less likely than the men to achieve a score of > or = 95 on the Barthel Activities of Daily Living Index (hazards ratio [HR] = 0.68; 95% confidence interval [CI], 0.52-0.90), carry out eight of nine instrumental activities of daily living without assistance (HR = 0.46; 95% CI, 0.30-0.68), and score > or = 90 on the SF-36 Health Survey physical functioning scale (HR = 0.54; 95% CI, 0.28-1.01). When age, prestroke physical functioning, stroke severity, and depressive status at baseline were controlled in the analysis, women in the study continued to be less likely (HR = 0.51; 95% CI, 0.32-0.79) than men in the study to be able to carry out eight of nine instrumental activities of daily living completely without assistance, but there were no observed sex differences in achievement of independence in basic activities of daily living or higher physical functioning. Prestroke physical functioning and depressive symptoms are important factors in the investigation of sex differences in stroke recovery. Lower recovery of activities of daily living and physical functioning in women after stroke may be due to multifactorial effects of older age, poor physical function prior to stroke onset, and depressive status after stroke.


Living longer with a greater health burden--changes in the burden of disease and injury in the Northern Territory Indigenous population between 1994-1998 and 1999-2003.
To measure changes over time in the burden of disease for Northern Territory (NT) Indigenous and non-Indigenous population. The numbers, and crude and age-adjusted rates of disability adjusted life years (DALY) were calculated for periods 1994-1998 and 1999-2003. A measure of information bias was developed to adjust for the tendency of years lost to disability (a component of DALY) to increase over time because of increasing data availability. The jackknife method was used for DALY uncertainty assessment. The all-cause DALY rate was stable for the non-Indigenous population, but increased for the Indigenous population. For both populations, the burden of premature death decreased while the burden of disability increased. For the Indigenous population, there were substantial increases in DALY rates for type 2 diabetes, depression, nephritis/nephrosis, suicide and sense organ disorders. The burden of disease for Indigenous people increased over the study periods, with improvement in the burden of fatal outcomes more than offset by substantial increase in the prevalence and severity of non-fatal conditions. The paradoxical shift of living longer with a greater health burden has not been previously reported for Indigenous Australians, and highlights the critical importance of prevention for sustaining life expectancy improvement and managing escalation of health costs. This study also demonstrated the usefulness of the DALY to monitor population health.


Ex vivo effects of corticosteroids on equine deep digital flexor and navicular fibrocartilage explant cell viability.
To investigate the effects of triamcinolone acetonide (TA) and methylprednisolone acetate (MPA) on the viability of resident cells within the fibrocartilage on the dorsal surface of the deep digital flexor tendon (FC-DDFT) and fibrocartilage on the flexor surface of the navicular bone (FC-NB) of horses. 12 to 14 explants of FC-DDFT and of FC-NB from grossly normal forelimbs of 5 cadavers of horses aged 9 to 15 years without evidence of musculoskeletal disease. Explants were incubated with culture medium (control) or TA-supplemented (0.6 or 6 mg/mL) or MPA-supplemented (0.5 or 5 mg/mL) medium for 6 or 24 hours. Explant metabolic activity and percentage of dead cells were assessed with a resazurin-based assay and live-dead cell staining, respectively, at each time point. Drug effects were assessed relative to findings for the respective control group. Application of TA (at both concentrations) did not significantly change the cell viability of FC-DDFT explants. For FC-NB explants, TA at 6 mg/mL significantly reduced the metabolic activity and increased the percentage of dead cells at both time points. With either MPA concentration, FC-DDFT and FC-NB explants had reduced metabolic activity and an increased percentage of dead cells at 24 hours, whereas only MPA at 5 mg/mL was cytotoxic at the 6-hour time point. In ex vivo explants, TA was less cytotoxic to equine FC-DDFT and FC-NB cells, compared with MPA. Further work is warranted to characterize the drugs' transcriptional and translational effects as well as investigate their cytotoxicity at lower concentrations.


Broadly neutralizing hemagglutinin stalk-specific antibodies require FcγR interactions for protection against influenza virus in vivo.
Neutralizing antibodies against influenza viruses have traditionally been thought to provide protection exclusively through their variable region; the contributions of mechanisms conferred by the Fc domain remain controversial. We investigated the in vivo contributions of Fc interactions with their cognate receptors for a collection of neutralizing anti-influenza antibodies. Whereas five broadly neutralizing monoclonal antibodies (bNAbs) targeting the conserved stalk region of hemagglutinin (HA) required interactions between the antibody Fc and Fc receptors for IgG (FcγRs) to confer protection from lethal H1N1 challenge, three strain-specific monoclonal Abs (mAbs) against the variable head domain of HA were equally protective in the presence or absence of FcγR interactions. Although all antibodies blocked infection, only anti-stalk bNAbs were capable of mediating cytotoxicity of infected cells, which accounts for their FcγR dependence. Immune complexes generated with anti-HA stalk mAb efficiently interacted with FcγRs, but anti-HA head immune complexes did not. These results suggest that FcγR binding capacity by anti-HA antibodies was dependent on the interaction of the cognate Fab with antigen. We exploited these disparate mechanisms of mAb-mediated protection to reengineer an anti-stalk bNAb to selectively enhance FcγR engagement to augment its protective activity. These findings reveal a previously uncharacterized property of bNAbs and guide an approach toward enhancing mAb-mediated antiviral therapeutics.


CenTime: Event-conditional modelling of censoring in survival analysis.
Survival analysis is a valuable tool for estimating the time until specific events, such as death or cancer recurrence, based on baseline observations. This is particularly useful in healthcare to prognostically predict clinically important events based on patient data. However, existing approaches often have limitations; some focus only on ranking patients by survivability, neglecting to estimate the actual event time, while others treat the problem as a classification task, ignoring the inherent time-ordered structure of the events. Additionally, the effective utilisation of censored samples-data points where the event time is unknown- is essential for enhancing the model's predictive accuracy. In this paper, we introduce CenTime, a novel approach to survival analysis that directly estimates the time to event. Our method features an innovative event-conditional censoring mechanism that performs robustly even when uncensored data is scarce. We demonstrate that our approach forms a consistent estimator for the event model parameters, even in the absence of uncensored data. Furthermore, CenTime is easily integrated with deep learning models with no restrictions on batch size or the number of uncensored samples. We compare our approach to standard survival analysis methods, including the Cox proportional-hazard model and DeepHit. Our results indicate that CenTime offers state-of-the-art performance in predicting time-to-death while maintaining comparable ranking performance. Our implementation is publicly available at https://github.com/ahmedhshahin/CenTime.


Impact of hypoglycemia in hospitalized patients.
Hypoglycemia is a common problem in hospitalized patients, particularly the elderly, frail, and severely ill. Hypoglycemia has been implicated in the development of adverse clinical outcomes, including increased mortality. Fear of iatrogenic hypoglycemia remains an obstacle to adequate inpatient glycemic control. However, evidence from large clinical trials is mixed: several intensive care unit studies have shown either reduced or no change in mortality with intensive glycemic control, despite high rates of iatrogenic hypoglycemia, and only 1 large study showed higher mortality. In the general ward setting, the association of hypoglycemia with worse outcomes and mortality has been frequently reported, but after multivariate adjustment for comorbidities this association disappears. Spontaneous hypoglycemia, rather than iatrogenic hypoglycemia, is strongly associated with mortality suggesting that hypoglycemia behaves as a biomarker rather than a causative factor of adverse outcomes. Inpatient glycemic management should be patient-centered, follow the current guidelines, and aimed at preventing hypoglycemia.


Spanish validation of Bad Sobernheim Stress Questionnaire (BSSQ (brace).es) for adolescents with braces.
As a result of scientific and medical professionals gaining interest in Stress and Health Related Quality of Life (HRQL), the aim of our research is, thus, to validate into Spanish the German questionnaire Bad Sobernheim Stress Questionnaire (BSSQ) (mit Korsett), for adolescents wearing braces. The methodology used adheres to literature on trans-cultural adaptation by doing a translation and a back translation; it involved 35 adolescents, ages ranging between 10 and 16, with Adolescent Idiopathic Scoliosis (AIS) and wearing the same kind of brace (Rigo System Chêneau Brace). The materials used were a socio-demographics data questionnaire, the SRS-22 and the Spanish version of BSSQ(brace).es. The statistical analysis calculated the reliability (test-retest reliability and internal consistency) and the validity (convergent and construct validity) of the BSSQ (brace).es. BSSQ(brace).es is reliable because of its satisfactory internal consistency (Cronbach's alpha coefficient was 0.809, p < 0.001) and temporal stability (test-retest method with a Pearson correlation coefficient of 0.902 (p < 0.01)).It demonstrated convergent validity with SRS-22 since the Pearson correlation coefficient was 0.656 (p < 0.01). By undertaking an Exploratory Principal Components Analysis, a latent structure was found based on two Components which explicate the variance at 60.8%. BSSQ (brace).es is reliable and valid and can be used with Spanish adolescents to assess the stress level caused by the brace.


ATLAS of Biochemistry: A Repository of All Possible Biochemical Reactions for Synthetic Biology and Metabolic Engineering Studies.
Because the complexity of metabolism cannot be intuitively understood or analyzed, computational methods are indispensable for studying biochemistry and deepening our understanding of cellular metabolism to promote new discoveries. We used the computational framework BNICE.ch along with cheminformatic tools to assemble the whole theoretical reactome from the known metabolome through expansion of the known biochemistry presented in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. We constructed the ATLAS of Biochemistry, a database of all theoretical biochemical reactions based on known biochemical principles and compounds. ATLAS includes more than 130 000 hypothetical enzymatic reactions that connect two or more KEGG metabolites through novel enzymatic reactions that have never been reported to occur in living organisms. Moreover, ATLAS reactions integrate 42% of KEGG metabolites that are not currently present in any KEGG reaction into one or more novel enzymatic reactions. The generated repository of information is organized in a Web-based database ( http://lcsb-databases.epfl.ch/atlas/ ) that allows the user to search for all possible routes from any substrate compound to any product. The resulting pathways involve known and novel enzymatic steps that may indicate unidentified enzymatic activities and provide potential targets for protein engineering. Our approach of introducing novel biochemistry into pathway design and associated databases will be important for synthetic biology and metabolic engineering.


Inactivation of steroid sulfatase by an active site-directed inhibitor, estrone-3-O-sulfamate.
Steroid sulfatases are responsible for the hydrolysis of 3beta-hydroxy steroid sulfates, such as cholesterol and pregnenolone sulfate, and have an important role in regulating the synthesis of estrogenic steroids, from estrone sulfate and dehydroepiandrosterone sulfate, in endocrine-dependent tumors. Although little is known about the mechanism by which the sulfate group is removed from a steroid nucleus, an active site-directed sulfatase inhibitor has been developed. This inhibitor, estrone-3-O-sulfamate (EMATE), was synthesized by treating the sodium salt of estrone with sulfamoyl chloride. This compound inhibited not only estrone sulfatase but also dehydroepiandrosterone sulfatase activity in placental microsomes and in intact MCF-7 breast cancer cells. Pretreatment of MCF-7 cells or placental microsomes with EMATE, followed by extensive washing or dialysis indicated irreversible inhibition. This was confirmed by showing that EMATE inhibited estrone sulfatase activity in placental microsomes in a time-, concentration-, and pH-dependent manner. The enzyme is protected from inactivation by estrone sulfate, which is also consistent with active site-directed inhibition. EMATE is proposed to inactivate estrone sulfatase by irreversible sulfamoylation of the enzyme. Maximum enzyme activity was detected at pH 8.6, and the maximum rate of enzyme inactivation by EMATE also occurred at this pH. The pKa values of the enzymatic reaction and pKa of inactivation were 7.2 and 9.8, providing evidence that two active site residues are being modified by EMATE. As the phenolic pKa of tyrosine (9.7) and the pKa of histidine will allow the roles that (6.8) are similar to the pKa values of inactivation, these amino acid residues may play a role in the catalytic mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)


Folic acid and vitamin B-12 supplementation does not favorably influence uracil incorporation and promoter methylation in rectal mucosa DNA of subjects with previous colorectal adenomas.
Adequate folate availability is necessary to sustain normal DNA synthesis and normal patterns of DNA methylation and these features of DNA can be modified by methylenetetrahydrofolate reductase (MTHFR) C677T genotype. This study investigated the effect of MTHFR C677T genotype and daily supplementation with 5 mg folic acid and 1.25 mg vitamin B-12 on uracil misincorporation into DNA and promoter methylation. Subjects (n = 86) with a history of colorectal adenoma and MTHFR CC or TT genotype were randomly assigned to receive folic acid plus vitamin B-12 or placebo for 6 mo. Uracil misincorporation and promoter methylation of 6 tumor suppressor and DNA repair genes were assessed in DNA from rectal biopsies at baseline and after the intervention. The biomarkers did not differ between the treated group and the placebo group after 6 mo compared with baseline. The uracil concentration of DNA increased in the treated group (5.37 fmol/microg DNA, P = 0.02), whereas it did not change in the placebo group (P = 0.42). The change from baseline of 4.01 fmol uracil/microg DNA tended to differ between the groups (P = 0.16). An increase in promoter methylation tended to occur more often in the intervention group than in the placebo group (OR = 1.67; P = 0.08). This study suggests that supplementation with high doses of folic acid and vitamin B-12 may not favorably influence uracil incorporation and promoter methylation in subjects with previous colorectal adenomas. Because such alterations may potentially increase the risk of neoplastic transformation, more research is needed to fully define the consequences of these molecular alterations.


Dissociable effects of acute antidepressant drug administration on subjective and emotional processing measures in healthy volunteers.
We have previously shown that single doses of serotonin-selective and noradrenaline-selective antidepressant agents produce positive biases in measures of emotional processing in healthy volunteers. The aim of the present study was to confirm and extend this finding by studying the effects of a single dose of the selective serotonin and noradrenaline re-uptake inhibitor, duloxetine. Healthy volunteers were randomly allocated to double-blind administration of either duloxetine 60 mg orally or placebo. Participants then completed a battery of emotional-processing tasks measuring facial expression recognition, emotional memory and emotion-potentiated startle. Subjective state was measured using visual analogue scales throughout the test period. Duloxetine enhanced the recognition of both disgusted and happy facial expressions and increased memory intrusions for positive personality characteristics in the free recall test. There were no significant effects on startle responses. However, duloxetine was not well tolerated and was associated with a high level of negative subjective effects. Despite the induction of negative subjective effects after duloxetine administration, some positive effects on emotional processing were seen in line with acute administration of serotonin-selective and noradrenaline-selective antidepressant agents. These results confirm the induction of fast changes in emotional processing in healthy volunteer groups and suggest a mechanism by which antidepressants may act in depression. Further studies are required to assess whether positive effects on emotional processing are more selective at a lower dose of duloxetine.


Epidemiology of alternaria alternata allergy: a prospective study in 6840 Italian asthmatic children.
The prevalence of Alternaria Alternata (AA), a mold causing in children severe asthma is scarcely known, also due to the underdiagnosis of AA allergy, frequently due to multiple sensitizations to molds. To analyze this issue we prospectively studied all children attending our Division between January 4, 1990 and December 31, 1997. A total of 6840 children with asthma or allergic rhinitis were evaluated. Diagnosis was established by family and personal history, physical examination, skin prick tests (SPTs) and RAST (Radio Allergo-Sorbent Test) for inhalants including AA. We further evaluated: (1) sensitization only to AA allergens without positivity for additional inhalants; (2) prevalence of AA positivity among children with asthma or allergic rhinitis; (3) concordance between SPTs and RAST for allergy to molds, (4) proportion of children treated with specific immunotherapy (SIT). Among the 6840 children 213 were positive to AA (3.3%), only 89/6840 children (1.3%) had AA monosensitization (p = 0.0001), a concordance between SPTs and RAST was present in 21/89 (23.6%) children (p = 0.0009), and only 9 children out of 89 were SIT treated. Concerning the clinical manifestations, 83 had asthma or allergic rhinitis, and 6 had asthma associated with atopic dermatitis. Family history was positive in 82.9% of children. The mean onset of AA sensitivity was at age 4 for males, and at age 5 for females. In childhood, AA allergy is a genetic affection. The SPT concordance with history and clinical examination appears to be operative. Due to life-threatening reactions in children with AA allergy, we suggest that those with suspected inhalant allergy be tested with AA allergens, and treated with SIT if positive.


Cigarette trafficking in five northeastern US cities.
Cigarette taxation is effective in reducing tobacco use in the USA. However, these benefits are reduced when taxes are unpaid. Cigarette trafficking (ie, the illegal importation of cigarettes into a high-tax jurisdiction from a lower-tax jurisdiction) is well documented in high-tax places like New York City (NYC), but the extent of trafficking in other northeastern cities is relatively unknown. To estimate the extent of cigarette trafficking in Boston, NYC, Philadelphia, Providence and Washington, DC, and project the benefits of reducing cigarette trafficking for recouping lost taxes and reducing smoking in these cities. Littered cigarette packs were collected from a random sample of Census tracts in five US cities. Data collection yielded 1439 total littered packs. The share of cigarette packs bearing proper local, known non-local, foreign or unknown, or no tax stamp was calculated for each city. These data were used to estimate tax revenue recovery if cigarette trafficking could be eliminated. We also estimated the extent to which eliminating cigarette trafficking would reduce cigarette consumption. Overall, 58.7% of packs did not have a proper local tax stamp, and 30.5-42.1% were attributed to trafficking. We estimate that eliminating cigarette trafficking would result in declines in youth smoking prevalence ranging from negligible in low-tax cities like Philadelphia to up to 9.3% in higher-tax NYC. We estimate that these five cities could recoup $680-729 million annually in cigarette tax revenue if cigarette trafficking was eliminated. Reducing cigarette trafficking would increase the effectiveness of tobacco taxes in reducing smoking and generate additional tax revenue, particularly in higher-taxed cities. Federal action to reduce cigarette trafficking, such as a track-and-trace system, is needed.


Neutrophil-derived serine proteinases enhance membrane type-1 matrix metalloproteinase-dependent tumor cell invasion.
Matrix metalloproteinase-2 degrades a variety of basement membrane components and is essential for tumor invasion. We have previously reported that membrane type-1 matrix metalloproteinase (MT1-MMP) cooperates with neutrophil-derived serine proteinases (NDPs; elastase, cathepsin G, protease-3) to activate matrix metalloproteinase-2. We therefore hypothesized that NDPs enhance tumor-cell invasion. Clones of human HT1080 fibrosarcoma cells transfected with MT1-MMP sense (HT-SE) or antisense CDNA (HT-AS) were used. These cells express either high (HT-SE) or extremely low levels (HT-AS) of MT1-MMP relative to nontransfected HT1080 cells (HT-WT). The cells were incubated in the presence or absence of purified NDP, with or without alpha 1-antitrypsin or the MMP inhibitor batimastat. Cell invasion was measured with the use of Boyden chambers with polycarbonate membranes coated with a reconstituted extracellular matrix. Under control conditions HT-WT and HT-SE cells were 4-fold more invasive than HT-AS cells. The addition of NDP increased HT-WT and HT-SE cell invasion 60% to 100% but had no effect on HT-AS cells. alpha 1-antitrypsin or batimastat did not decrease the baseline invasiveness of HT-WT and HT-SE cells; however, they abrogated the stimulatory effect of NDP. HT1080 cell invasion depends on MT1-MMP expression. MT1-MMP overexpression does not increase invasiveness by itself. NDPs increase invasion by MT1-MMP expressing cells by activating matrix metalloproteinase-2.


Identification of recent hybridization between gray wolves and domesticated dogs by SNP genotyping.
The ability to detect recent hybridization between dogs and wolves is important for conservation and legal actions, which often require accurate and rapid resolution of ancestry. The availability of a genetic test for dog-wolf hybrids would greatly support federal and legal enforcement efforts, particularly when the individual in question lacks prior ancestry information. We have developed a panel of 100 unlinked ancestry-informative SNP markers that can detect mixed ancestry within up to four generations of dog-wolf hybridization based on simulations of seven genealogical classes constructed following the rules of Mendelian inheritance. We establish 95 % confidence regions around the spatial clustering of each genealogical class using a tertiary plot of allele dosage and heterozygosity. The first- and second-backcrossed-generation hybrids were the most distinct from parental populations, with >90 % correctly assigned to genealogical class. In this article we provide a tool kit with population-level statistical quantification that can detect recent dog-wolf hybridization using a panel of dog-wolf ancestry-informative SNPs with divergent allele frequency distributions.


Phylogenetic and functional evidence suggests that deep-ocean ecosystems are highly sensitive to environmental change and direct human disturbance.
An understanding of the balance of interspecific competition and the physical environment in structuring organismal communities is crucial because those communities structured primarily by their physical environment typically exhibit greater sensitivity to environmental change than those structured predominantly by competitive interactions. Here, using detailed phylogenetic and functional information, we investigate this question in macrofaunal assemblages from Northwest Atlantic Ocean continental slopes, a high seas region projected to experience substantial environmental change through the current century. We demonstrate assemblages to be both phylogenetically and functionally under-dispersed, and thus conclude that the physical environment, not competition, may dominate in structuring deep-ocean communities. Further, we find temperature and bottom trawling intensity to be among the environmental factors significantly related to assemblage diversity. These results hint that deep-ocean communities are highly sensitive to their physical environment and vulnerable to environmental perturbation, including by direct disturbance through fishing, and indirectly through the changes brought about by climate change.


Poplar carbohydrate-active enzymes: whole-genome annotation and functional analyses based on RNA expression data.
Carbohydrate-active enzymes (CAZymes) catalyze the formation and modification of glycoproteins, glycolipids, starch, secondary metabolites and cell wall biopolymers. They are key enzymes for the biosynthesis of food and renewable biomass. Woody biomass is particularly important for long-term carbon storage and as an abundant renewable natural resource for many industrial applications. This study presents a re-annotation of CAZyme genes in the current Populus trichocarpa genome assembly and in silico functional characterization, based on high-resolution RNA-Seq data sets. Altogether, 1914 CAZyme and expansin genes were annotated in 101 families. About 1797 of these genes were found expressed in at least one Populus organ. We identified genes involved in the biosynthesis of different cell wall polymers and their paralogs. Whereas similar families exist in poplar and Arabidopsis thaliana (with the exception of CBM13 found only in poplar), a few families had significantly different copy numbers between the two species. To identify the transcriptional coordination and functional relatedness within the CAZymes and other proteins, we performed co-expression network analysis of CAZymes in wood-forming tissues using the AspWood database (http://aspwood.popgenie.org/aspwood-v3.0/) for Populus tremula. This provided an overview of the transcriptional changes in CAZymes during the transition from primary to secondary wall formation, and the clustering of transcripts into potential regulons. Candidate enzymes involved in the biosynthesis of polysaccharides were identified along with many tissue-specific uncharacterized genes and transcription factors. These collections offer a rich source of targets for the modification of secondary cell wall biosynthesis and other developmental processes in woody plants.


High-power diode-end-pumped laser with multi-segmented Nd-doped yttrium vanadate.
A Nd:YVO4 crystal consisting of three segments with different doping concentrations is originally developed for power scaling in diode-end-pumped solid-state laser. We systematically make a comparison of laser characteristics between the multi-segmented and conventional composite crystals to show the feasibility of using gain medium with multiple doping concentrations for power scale-up in end-pumped laser without introducing significant thermally accompanied effects. We further construct a dual-end-pumped actively Q-switched oscillator at 1064 nm to verify the usefulness of our crystal design, where the largest pulse energy of 1.06 mJ and the highest output power of 45 W are efficiently generated.


Improved survival after heart-lung transplantation.
Sixty-two patients underwent 65 heart-lung transplant operations before April 1989. Thirty of these patients (operated on March 1981 to February 1986) were given cyclosporine and prednisone for immunosuppression. These 30 patients (group 1) are compared with the 32 most recent patients (group 2) treated with cyclosporine, prednisone, and azathioprine. Patient characteristics (group 1 versus group 2) were similar including age (mean 32 versus 26 years) and indication for operation (51% versus 38%, primary pulmonary hypertension; 46% versus 45%, Eisenmenger complex; 3% versus 17%, other). The perioperative (in-hospital) mortality rate was 35% in group 1 versus 16% in group 2. Obliterative bronchiolitis developed in 12 (63%) of 19 hospital survivors in group 1 from 2 months to 49 months postoperatively (mean 12 months). This complication has developed in five (20%) of the 25 hospital survivors in group 2 from 3 to 13 months postoperatively (mean 8 months). Overall, 24 of the 30 patients in group 1 died (infection in eight, obliterative bronchiolitis in five, graft coronary artery disease in two, other causes in nine). Eight of the 32 patients in group 2 died (infection in six, other cases in two). Survival rates for group 1 patients were 60% at 1 year, 50% at 2 years, 43% at 3 years, and 25% at 5 years. Survival rates for group 2 patients were 73% at 1 year, 73% at 2 years, and 65% at 3 years (p less than 0.05). When group 2 patients were compared with 134 patients undergoing heart transplantation during the same time period, there was no difference in survival. Routine bronchoscopic surveillance in the more recent group of patients has led to the earlier detection of rejection and infection. In combination with decreased perioperative mortality and augmented immunosuppression, this had led to improved survival in our more recent heart-lung transplant recipients. In addition, the earlier recognition of infection and rejection has led to a decrease in severity of obliterative bronchiolitis.


Optimized interstitial PDT prostate treatment planning with the Cimmino feasibility algorithm.
The primary aim of this study was to determine whether optimized photodynamic therapy (PDT) treatment planning (seeking optimized positions, lengths, and strengths of the light sources to satisfy a given dose prescription) can improve dose coverage to the prostate and the sparing of critical organs relative to what can be achieved by the standard PDT plan. The Cimmino algorithm and search procedures based on that algorithm were tested for this purpose. A phase I motexafin lutetium (MLu)-mediated photodynamic therapy protocol is ongoing at the University of Pennsylvania. PDT for the prostate is performed with cylindrical diffusing fibers of various lengths inserted perpendicular to a base plate to obtain longitudinal coverage by a matrix of parallel catheters. The standard plan for the protocol uses sources of equal strength with equal spaced (1-cm) loading. Uniform optical properties were assumed. Our algorithms produce plans that cover the prostate and spare the urethra and rectum with less discrepancy from the dose prescription than the standard plan. The Cimmino feasibility algorithm is fast enough that changes to the treatment plan may be made in the operating room before and during PDT to optimize light delivery.


Fractures of the capitellum and trochlea.
Recent work has established that apparently isolated fractures of the capitellum are often more complex and involve the lateral epicondyle, trochlea, and posterior aspect of the distal part of the humerus. We assessed the experience with operative stabilization of fractures of the capitellum and trochlea at one level-I trauma center over a twenty-eight-year period. Thirty classifiable partial articular fractures involving the capitellum and trochlea were included in the study. Twenty-seven patients were followed for a minimum of twelve months, and fourteen patients returned for long-term follow-up at a median of seventeen years. The early and long-term results were evaluated according to the Broberg and Morrey Functional Rating Index. The long-term results were also evaluated according to the Mayo Elbow Performance Index (MEPI), the American Shoulder and Elbow Surgeons (ASES) score, and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire. Eighteen patients (67%) had one or more subsequent surgical procedures, and eight of these patients had the procedure to address surgical complications. Five of the eight patients with complications and ten additional patients underwent routine removal of implants; these fifteen patients included twelve of the fourteen patients in the long-term cohort. In addition to the fracture of the distal part of the humerus, four patients had a dislocation of the elbow; three, a fracture of the olecranon or the proximal part of the ulna; and two, a fracture of the radial head. The median arc of flexion improved from 106 degrees at the time of early follow-up to 119 degrees at the time of long-term follow-up (p < 0.05). In the group of fourteen patients with long-term follow-up, the median Broberg and Morrey score was 93 points at the time of early follow-up and 95 points at the time of late follow-up. The functional results were worse for patients with a Type-3 fracture, as classified with the system of Dubberley et al., than they were for those with a Type-1 fracture. The fourteen patients with long-term follow-up had a median MEPI of 98 points, a median ASES score of 88 points, and a median DASH score of 8 points; nine of the fourteen patients had radiographic signs of arthrosis. The vast majority of what appear to be capitellar fractures are actually complex fractures of the articular surface involving both the capitellum and the trochlea. More complex fractures have worse functional results; however, the functional results of operative treatment seem to be durable over time.


Safe water treatment and storage in the home. A practical new strategy to prevent waterborne disease.
In many parts of the developing world, drinking water is collected from unsafe surface sources outside the home and is then held in household storage vessels. Drinking water may be contaminated at the source or during storage; strategies to reduce waterborne disease transmission must safeguard against both events. We describe a two-component prevention strategy, which allows an individual to disinfect drinking water immediately after collection (point-of-use disinfection) and then to store the water in narrow-mouthed, closed vessels designed to prevent recontamination (safe storage). New disinfectant generators and better storage vessel designs make this strategy practical and inexpensive. This approach empowers households and communities that lack potable water to protect themselves against a variety of waterborne pathogens and has the potential to decrease the incidence of waterborne diarrheal disease.


Differential effects of exogenous and autocrine growth hormone on LNCaP prostate cancer cell proliferation and survival.
The prostate gland is regulated by multiple hormones and growth factors that may also affect prostate tumorigenesis. Growth hormone (GH) contributes to prostate development and function, but the direct effects of GH on prostate cancer cells are not well understood. The expression of endogenous GH in prostate cancer cell lines has also been observed, suggesting the potential for an effect of autocrine GH. In the present study, we measure the levels of GH and GH receptor (GHR) mRNA in multiple prostate cancer and normal prostate-derived cell lines, and compare the effects of exogenous and autocrine GH on LNCaP prostate cancer cell proliferation and apoptosis, and the associated signal transduction pathways. We found that GHR and GH expression were higher in the prostate cancer cell lines, and that exogenous GH increased LNCaP cell proliferation, but had no effect on apoptosis. In contrast, autocrine GH overexpression reduced LNCaP cell proliferation and increased apoptosis. The distinct actions of exogenous and autocrine GH were accompanied by differences in the involvement of GHR-associated signal transduction pathways, and were paralleled by an alteration in the subcellular localization of GHR, in which autocrine GH appeared to sequester GHR in the Golgi and endoplasmic reticulum. This alteration of GHR trafficking may underlie a distinct mode of GH-mediated signaling associated with the effect of autocrine GH. These findings clarify the potential effects of GH on prostate cancer cell function, and indicate that the activity of autocrine GH may be distinct from that of endocrine GH in prostate cancer cells.


Revision of cyrtandra (gesneriaceae) in the marquesas islands.
During the preparation of the Vascular Flora of the Marquesas Islands three new species of Cyrtandra (Gesneriaceae) have come to light and are described herein: C. uapouensis W. L. Wagner & Lorence, C. uahukaensis W. L. Wagner & Lorence, and C. kenwoodii W. L. Wagner & A. J. Wagner. Amended descriptions of the eight previously described Marquesan species are also provided as well as a key to the species. With the description of these the new species Cyrtandra in the Marquesas Islands consists of 11 species, six of which have been included in recent molecular phylogenetic studies of Pacific Cyrtandra, and appear to have arisen from one original introduction. If the other five species are members of this Marquesas clade then Cyrtandra would represent the largest lineage of Marquesas vascular plants. Psychotria is largest genus in the Marquesas Islands with 13 species, but is thought to consist of three separate lineages.


Energetic driving forces are maintained in resting rat skeletal muscle after dietary creatine supplementation.
The total creatine (TCr) pool of skeletal muscle is composed of creatine (Cr) and phosphocreatine (PCr). In resting skeletal muscle, the ratio of PCr to TCr (PCr/TCr; PCr energy charge) is approximately 0.6-0.8, depending on the fiber type. PCr/TCr is linked to the cellular free energy of ATP hydrolysis by the Cr kinase equilibrium. Dietary Cr supplementation increases TCr in skeletal muscle. However, many previous studies have reported data indicating that PCr/TCr falls after supplementation, which would suggest that Cr supplementation alters the resting energetic state of myocytes. This study investigated the effect of Cr supplementation on the energy phosphates of resting skeletal muscle. Male rats were fed either rodent chow (control) or chow supplemented with 2% (wt/wt) Cr. After 2 wk on the diet, the gastrocnemius and soleus muscles were freeze clamped and removed from anesthetized animals. Cr supplementation increased TCr, PCr, and Cr levels in the gastrocnemius by 20, 22, and 17%, respectively (P < 0.05). A numerical 6% higher mean soleus TCr in Cr-supplemented rats was not statistically significant. All other energy phosphate concentrations, free energy of ATP hydrolysis, and PCr/TCr were not different between the two groups in either muscle. We conclude that Cr supplementation simply increased TCr in fast-twitch rat skeletal muscle but did not otherwise alter resting cellular energetic state.


A review of checkpoint inhibitors in the management of renal cell carcinoma.
Renal cell carcinoma is a common malignancy of the genitourinary system and is the eight most common cancer type in the United States. The overall incidence of renal cell carcinoma appears to be increasing but death rates have been declining. Patients with poor risk, advanced disease have a two-year survival rate of approximately 7%. Prior to the advent of tyrosine kinase inhibitors, anti-vascular endothelial growth factor antibodies, mammalian target of rapamycin inhibitors, and checkpoint inhibitors, IFN-α and high-dose IL-2, were standard of care treatment options but, conversely, their use is now limited to select patients. Immunotherapies have played a significant role in the treatment of various cancers and have continued to expand. It is of utmost importance that practitioners include checkpoint inhibitors as treatment options for renal cell carcinoma as they mark a new era in the treatment of advanced or relapsed setting. Nivolumab, pembrolizumab, avelumab, ipilimumab, and atezolizumab all play a role in management of disease as either monotherapy or in combination with other agents. Ongoing clinical trials are ongoing to further assess the benefits of inducing cellular immunity in the treatment of renal cell carcinoma. In this article, the available data on immune checkpoint inhibitors for the treatment of advanced or relapsed renal cell carcinoma and their place in therapy are reviewed.


Changes in plasma homovanillic acid concentrations in schizophrenic patients following neuroleptic discontinuation.
Changes in plasma levels of the dopamine metabolite homovanillic acid have been reported to correlate with changes in the severity of schizophrenic symptoms during neuroleptic administration and after neuroleptic discontinuation. This study examined the effects of discontinuation of neuroleptic treatment on plasma homovanillic acid levels in 23 patients with chronic schizophrenia. It was hypothesized that clinical decompensation would be associated with increased plasma homovanillic acid levels. Plasma homovanillic acid was measured during administration of neuroleptic medication and during a subsequent 6-week drug-free period. Nine patients decompensated during the drug-free period and 14 patients did not. Following drug discontinuation, plasma homovanillic acid concentrations were higher in schizophrenic patients who decompensated than in those who did not. Furthermore, peak plasma homovanillic acid elevation after discontinuation of neuroleptic medication was significantly correlated with peak Brief Psychiatric Rating Scale increase. The data suggest that, in some schizophrenic patients, symptomatic decompensation after discontinuation of neuroleptic treatment is associated with increases in dopamine turnover.


Hyperkalemia alters EDHF-mediated hyperpolarization and relaxation in coronary arteries.
Hyperkalemic solutions are widely used to preserve organs for transplantation and for cardiac surgery. The present study was designed to test the hypothesis that hyperkalemia may alter endothelial function through a non-nitric oxide (NO) pathway, since preliminary studies have shown that the NO pathway may not be affected. Porcine coronary artery rings were studied in organ chambers. After incubation with 20 or 50 mM K+ for 1 h, the indomethacin- and NG-nitro-L-arginine+ (L-NNA)-resistant relaxation induced by A23187 or bradykinin, which could be further inhibited by tetraethylammonium but not glibenclamide, was significantly reduced. Incubation with hyperkalemia also significantly increased the concentration eliciting 50% of the maximal response to A23187 and bradykinin. A23187-induced hyperpolarization of the membrane potential was significantly reduced by hyperkalemic incubation. However, 1-h incubation with hyperkalemia does not affect the endothelial Ca2+ concentration. We conclude that exposure to hyperkalemia reduces the indomethacin- and L-NNA-resistant endothelium-dependent relaxation and endothelium-dependent hyperpolarization. This reduction in the relaxation and hyperpolarization is related to the endothelium-derived hyperpolarizing factor by affecting its effect on the smooth muscle cell, probably through partially depolarizing the membrane, and the Ca2(+)- activated K+ channels rather than by affecting its biosynthesis and/or release in the endothelial cell. Our study may suggest a new mechanism for coronary dysfunction after exposure to hyperkalemic cardioplegia and organ preservation solutions.


A novel tissue culture model for evaluating the effect of aging on stem cell fate in adult microvascular networks.
In vitro models of angiogenesis are valuable tools for understanding the underlying mechanisms of pathological conditions and for the preclinical evaluation of therapies. Our laboratory developed the rat mesentery culture model as a new tool for investigating mechanistic cell-cell interactions at specific locations across intact blood and lymphatic microvascular networks ex vivo. The objective of this study was to report a method for evaluating the effect of aging on human stem cell differentiation into pericytes during angiogenesis in cultured microvascular networks. DiI labeled exogenous stem cells were seeded onto harvested adult Wistar rat mesenteric tissues and cultured in alpha-MEM + 1% serum for up to 5 days according to four experimental groups: (1) adult human adipose-derived stem cells (hASCs), (2) aged hASCs, (3) adult human bone marrow-derived stem cells (hBMSCs), and (4) aged hBMSCs. Angiogenesis per experimental group was supported by observation of increased vessel density and capillary sprouting. For each tissue per experimental group, a subset of cells was observed in typical pericyte location wrapped along blood vessels. Stem cell differentiation into pericytes was supported by the adoption of elongated pericyte morphology along endothelial cells and positive NG2 labeling. The percentage of cells in pericyte locations was not significantly different across the experimental groups, suggesting that aged mesenchymal stem cells are able to retain their differentiation capacity. Our results showcase an application of the rat mesentery culture model for aging research and the evaluation of stem cell fate within intact microvascular networks.


Sequence logos: a new way to display consensus sequences.
A graphical method is presented for displaying the patterns in a set of aligned sequences. The characters representing the sequence are stacked on top of each other for each position in the aligned sequences. The height of each letter is made proportional to its frequency, and the letters are sorted so the most common one is on top. The height of the entire stack is then adjusted to signify the information content of the sequences at that position. From these 'sequence logos', one can determine not only the consensus sequence but also the relative frequency of bases and the information content (measured in bits) at every position in a site or sequence. The logo displays both significant residues and subtle sequence patterns.


Therapeutic effect of Lactobacillus rhamnosus SHA113 on intestinal infection by multi-drug-resistant Staphylococcus aureus and its underlying mechanisms.
Staphylococcus aureus, especially multi-drug-resistant (MDR) pathogenic S. aureus, poses a severe threat to food safety and human health. Probiotics offer promising potential for the control of MDR pathogens because of their safe and biofunctional properties. This study shows that Lactobacillus rhamnosus SHA113, a strain isolated from the milk of healthy women, could efficiently inhibit MDR S. aureus both in vitro and in vivo. In vitro, L. rhamnosus efficiently inhibited and even killed drug resistant and drug sensitive S. aureus strains. In vivo experiments showed that SHA113 could efficiently decrease the number of S. aureus cells, inhibit the expression of inflammatory factors TNF-α and IL-6, and restore the level of white cells and neutrophils in the blood. SHA113 could also efficiently repair damage of the intestinal barrier and other functions impaired by S. aureus infection. This was indicated by a change of intestinal villi length and structure, and an up-regulated expression of tight junction proteins ZO-1 and occludin. SHA113 also restored the structural damage of immune organs, such as the enlargement of the spleen and the increased level of inflammatory cytokines caused by S. aureus infection. More importantly, L. rhamnosus SHA113 showed more effective inhibitory and therapeutic effects on MDR S. aureus strain ZBQ006 than on drug sensitive S. aureus strain 29213. These results illustrated that L. rhamnosus SHA113 has great potential for the treatment of MDR S. aureus contamination as food control and for therapeutic treatment.


Effect of conventional and microwave glazing on surface roughness of metal ceramics: An atomic force microscopy analysis.
This study investigated and compared the surface roughness achieved by glazing porcelain samples in a conventional and a microwave oven. Two commercial brands of metal ceramics were used, VITA VMK MASTER and IPS CLASSIC. Sixty samples were fabricated, 30 for each type of ceramic. The samples were sintered in the conventional oven and hand-polished to remove any irregularities. Samples (n = 10) from each type of ceramic were further divided into three groups as follows: hand-polished (Group A), conventional oven glazed (Group B), and microwave glazed (Group C). Each specimen was evaluated for surface roughness by atomic force microscope. Data were statistically analyzed using two-way analysis of variance (ANOVA) and Tukey's post hoc test (a = 0.05). Two-way ANOVA indicated a highly significant difference in surface roughness based on the type of glazing (P < 0.001), there was a significant difference based on the metal ceramics (P = 0.002). There was also a significant interaction between the type of glazing and metal ceramics (P = 0.009). The images obtained from the atomic force microscope corroborated the measured values. All the results indicate that microwave glazing can be a feasible option for glazing porcelain specimens. It was concluded that surface topography is influenced by surface treatment and microwave glazed ceramic is superior to conventional oven glazed ceramic and hand-polishing showed greater surface roughness when compared to glazing. IPS CLASSIC ceramic showed relatively smooth surface when compared to VITA VMK MASTER irrespective of the surface treatment.


Improved staff procedure skills lead to improved managment skills: an observational study in an educational setting.
Good staff procedure skills in a management group during incidents and disasters are believed to be a prerequisite for good management of the situation. However, this has not been demonstrated scientifically. Templates for evaluation results from performance indicators during simulation exercises have previously been tested. The aim of this study was to demonstrate the possibility that these indicators can be used as a tool for studying the relationship between good management skills and good staff procedure skills. Good and structured work (staff procedure skills) in a hospital management group during simulation exercises in disaster medicine is related to good and timely decisions (good management skills). Results from 29 consecutive simulation exercises in which staff procedure skills and management skills were evaluated using quantitative measurements were included. The statistical analysis method used was simple linear regression with staff procedure skills as the response variable and management skills as the predictor variable. An overall significant relationship was identified between staff procedure skills and management skills (p(2)0.05). This study suggests that there is a relationship between staff procedure skills and management skills in the educational setting used. Future studies are needed to demonstrate if this also can be observed during actual incidents.


Human AlkB homologue 5 is a nuclear 2-oxoglutarate dependent oxygenase and a direct target of hypoxia-inducible factor 1α (HIF-1α).
Human 2-oxoglutarate oxygenases catalyse a range of biological oxidations including the demethylation of histone and nucleic acid substrates and the hydroxylation of proteins and small molecules. Some of these processes are centrally involved in regulation of cellular responses to hypoxia. The ALKBH proteins are a sub-family of 2OG oxygenases that are defined by homology to the Escherichia coli DNA-methylation repair enzyme AlkB. Here we report evidence that ALKBH5 is probably unique amongst the ALKBH genes in being a direct transcriptional target of hypoxia inducible factor-1 (HIF-1) and is induced by hypoxia in a range of cell types. We show that purified recombinant ALKBH5 is a bona fide 2OG oxygenase that catalyses the decarboxylation of 2OG but appears to have different prime substrate requirements from those so far defined for other ALKBH family members. Our findings define a new class of HIF-transcriptional target gene and suggest that ALKBH5 may have a role in the regulation of cellular responses to hypoxia.


A Plant Stem Inoculation Assay for Assessing Transmission of Phytophthora infestans from Potato Seed Tubers to Emerged Shoots.
A stem inoculation assay was developed to assess transmission of Phytophthora infestans from the base of belowground potato stems to emerged aboveground shoots. Sporangia of P. infestans subsequently developed on aboveground stems after belowground stems were inoculated with isolates BF05 (US 8 genotype), WA10.1 (US 24), and 110B (US 11) by (i) placing a filter paper square saturated with a spore suspension onto nonwounded belowground stem tissue, (ii) placing a filter paper square saturated with a spore suspension onto wounded belowground stem tissue, and (iii) dipping the base of the cut belowground stem into a spore suspension. Plant stems were then incubated in a humidity chamber at 10 to 21.5°C for 7 to 28 days. Incidence of aboveground stems with sporulation was significantly less when stems were inoculated with isolate 110B than with isolates BF05 and WA10.1 but length of latent period did not differ among the three isolates. Incidence of aboveground stems with sporangia was significantly (P < 0.05) greater for 'Russet Norkotah' than for 'Umatilla Russet' when inoculated near the point of seed piece attachment. Incidence of aboveground stems with sporulation did not differ among incubation temperatures of 10, 15, and 21.5°C. Latent period significantly increased as temperature decreased. Age of stem did not appear to affect incidence of transmission. Incidence of transmission of P. infestans to aboveground stems from the base of belowground stems was significantly lowered when aboveground stems were treated with selected fungicides 1 h before inoculation.


Meta-analysis of transcutaneous electrical nerve stimulation for relief of spinal pain.
We conducted a systematic review and meta-analysis analysing the existing data on transcutaneous electrical nerve stimulation (TENS) or interferential current (IFC) for chronic low back pain (CLBP) and/or neck pain (CNP) taking into account intensity and timing of stimulation, examining pain, function and disability. Seven electronic databases were searched for TENS or IFC treatment in non-specific CLBP or CNP. Four reviewers independently selected randomized controlled trials (RCTs) of TENS or IFC intervention in adult individuals with non-specific CLBP or CNP. Primary outcomes were for self-reported pain intensity and back-specific disability. Two reviewers performed quality assessment, and two reviewers extracted data using a standardized form. Nine RCTs were selected (eight CLBP; one CNP), and seven studies with complete data sets were included for meta-analysis (655 participants). For CLBP, meta-analysis shows TENS/IFC intervention, independent of time of assessment, was significantly different from placebo/control (p < 0.02). TENS/IFC intervention was better than placebo/control, during therapy (p = 0.02), but not immediately after therapy (p = 0.08), or 1-3 months after therapy (p = 0.99). Analysis for adequate stimulation parameters was not significantly different, and there was no effect on disability. This systematic review provides inconclusive evidence of TENS benefits in low back pain patients because the quality of the studies was low, and adequate parameters and timing of assessment were not uniformly used or reported. Without additional high-quality clinical trials using sufficient sample sizes and adequate parameters and outcome assessments, the outcomes of this review are likely to remain unchanged. These data highlight the need for additional high-quality RCTs to examine the effects of TENS in CLBP. Trials should consider intensity of stimulation, timing of outcome assessment and assessment of pain, disability and function.


Extinction in a hyperdiverse endemic Hawaiian land snail family and implications for the underestimation of invertebrate extinction.
The International Union for Conservation of Nature (IUCN) Red List includes 832 species listed as extinct since 1600, a minuscule fraction of total biodiversity. This extinction rate is of the same order of magnitude as the background rate and has been used to downplay the biodiversity crisis. Invertebrates comprise 99% of biodiversity, yet the status of a negligible number has been assessed. We assessed extinction in the Hawaiian land snail family Amastridae (325 species, IUCN lists 33 as extinct). We did not use the stringent IUCN criteria, by which most invertebrates would be considered data deficient, but a more realistic approach comparing historical collections with modern surveys and expert knowledge. Of the 325 Amastridae species, 43 were originally described as fossil or subfossil and were assumed to be extinct. Of the remaining 282, we evaluated 88 as extinct and 15 as extant and determined that 179 species had insufficient evidence of extinction (though most are probably extinct). Results of statistical assessment of extinction probabilities were consistent with our expert evaluations of levels of extinction. Modeling various extinction scenarios yielded extinction rates of 0.4-14.0% of the amastrid fauna per decade. The true rate of amastrid extinction has not been constant; generally, it has increased over time. We estimated a realistic average extinction rate as approximately 5%/decade since the first half of the nineteenth century. In general, oceanic island biotas are especially susceptible to extinction and global rate generalizations do not reflect this. Our approach could be used for other invertebrates, especially those with restricted ranges (e.g., islands), and such an approach may be the only way to evaluate invertebrates rapidly enough to keep up with ongoing extinction.


Cranberry proanthocyanidins inhibit esophageal adenocarcinoma in vitro and in vivo through pleiotropic cell death induction and PI3K/AKT/mTOR inactivation.
Cranberries are rich in bioactive constituents known to improve urinary tract health and more recent evidence supports cranberries possess cancer inhibitory properties. However, mechanisms of cancer inhibition by cranberries remain to be elucidated, particularly in vivo. Properties of a purified cranberry-derived proanthocyanidin extract (C-PAC) were investigated utilizing acid-sensitive and acid-resistant human esophageal adenocarcinoma (EAC) cell lines and esophageal tumor xenografts in athymic NU/NU mice. C-PAC induced caspase-independent cell death mainly via autophagy and low levels of apoptosis in acid-sensitive JHAD1 and OE33 cells, but resulted in cellular necrosis in acid-resistant OE19 cells. Similarly, C-PAC induced necrosis in JHAD1 cells pushed to acid-resistance via repeated exposures to an acidified bile cocktail. C-PAC associated cell death involved PI3K/AKT/mTOR inactivation, pro-apoptotic protein induction (BAX, BAK1, deamidated BCL-xL, Cytochrome C, PARP), modulation of MAPKs (P-P38/P-JNK) and G2-M cell cycle arrest in vitro. Importantly, oral delivery of C-PAC significantly inhibited OE19 tumor xenograft growth via modulation of AKT/mTOR/MAPK signaling and induction of the autophagic form of LC3B supporting in vivo efficacy against EAC for the first time. C-PAC is a potent inducer of EAC cell death and is efficacious in vivo at non-toxic behaviorally achievable concentrations, holding promise for preventive or therapeutic interventions in cohorts at increased risk for EAC, a rapidly rising and extremely deadly malignancy.


Multiorder correction algorithms to remove image distortions from mass spectrometry imaging data sets.
Time-of-flight secondary ion mass spectrometry imaging is a rapidly evolving technology. Its main application is the study of the distribution of small molecules on biological tissues. The sequential image acquisition process remains susceptible to measurement distortions that can render imaging data less analytically useful. Most of these artifacts show a repetitive nature from tile to tile. Here we statistically describe these distortions and derive two different algorithms to correct them. Both a generalized linear model approach and the linear discriminant analysis approach are able to increase image quality for negative and positive ion mode data sets. Additionally, performing simulation studies with repetitive and nonrepetitive tiling error we show that both algorithms are only removing repetitive distortions. It is further shown that the spectral component of the data set is not altered by the use of these correction methods. Both algorithms presented in this work greatly increase the image quality and improve the analytical usefulness of distorted images dramatically.


Enhancement of the supercapacitive properties of laser deposited graphene-based electrodes through carbon nanotube loading and nitrogen doping.
Several technological routes are being investigated for improving the energy storage capability and power delivery of electrochemical capacitors. In this work, ternary hybrid electrodes composed of conducting graphene/reduced graphene oxide (rGO), which store charge mainly through electric double-layer mechanisms, covered by NiO nanostructures, for adding pseudocapacitance, were fabricated through a matrix assisted pulsed laser evaporation technique. The incorporation of multiwall carbon nanotubes (MWCNTs) provokes an increase of the porosity and thus, a substantial enhancement of the electrodes' capacitance (from 4 to 20 F cm-3 at 10 mV s-1). Volumetric capacitances of 34 F cm-3 were also obtained with electrodes containing just carbon nanotubes coated with NiO nanostructures. Moreover, the use of nitrogen containing precursors (ammonia, urea) for laser-induced N-doping of the nanocarbons also provokes a notable increase of the capacitance. Remarkably, N-containing groups in rGO-MWCNTs mainly add electric double layer charge storage, pointing to an increase of electrode porosity, whereas redox reactions contribute with a minor diffusion fraction. It was also observed that the loading of carbon nanotubes leads to an increase of diffusion-controlled charge storage mechanisms versus capacitive ones in rGO-based electrodes, the opposite effect being observed in graphene electrodes.


Trends in Canadian hospital standardised mortality ratios and palliative care coding 2004-2010: a retrospective database analysis.
The hospital standardised mortality ratio (HSMR), anchored at an average score of 100, is a controversial macromeasure of hospital quality. The measure may be dependent on differences in patient coding, particularly since cases labelled as palliative are typically excluded. To determine whether palliative coding in Canada has changed since the 2007 national introduction of publicly released HSMRs, and how such changes may have affected results. Retrospective database analysis. Inpatients in Canadian hospitals from April 2004 to March 2010. 12 593 329 hospital discharges recorded in the Canadian Institute for Health Information (CIHI) Discharge Abstract Database from April 2004 to March 2010. Crude mortality and palliative care coding rates. HSMRs calculated with the same methodology as CIHI. A derived hospital standardised palliative ratio (HSPR) adjusted to a baseline average of 100 in 2004-2005. Recalculated HSMRs that included palliative cases under varying scenarios. Crude mortality and palliative care coding rates have been increasing over time (p<0.001), in keeping with the nation's advancing overall morbidity. HSMRs in 2008-2010 were significantly lower than in 2004-2006 by 8.55 points (p<0.001). The corresponding HSPR rises dramatically between these two time periods by 48.83 points (p<0.001). Under various HSMR scenarios that included palliative cases, the HSMR would have at most decreased by 6.35 points, and may have even increased slightly. Inability to calculate a definitively comparable HSMR that include palliative cases and to account for closely timed changes in national palliative care coding guidelines. Palliative coding rates in Canadian hospitals have increased dramatically since the public release of HSMR results. This change may have partially contributed to the observed national decline in HSMR.


Me, myself, bye: regional alterations in glutamate and the experience of ego dissolution with psilocybin.
There is growing interest in the therapeutic utility of psychedelic substances, like psilocybin, for disorders characterized by distortions of the self-experience, like depression. Accumulating preclinical evidence emphasizes the role of the glutamate system in the acute action of the drug on brain and behavior; however this has never been tested in humans. Following a double-blind, placebo-controlled, parallel group design, we utilized an ultra-high field multimodal brain imaging approach and demonstrated that psilocybin (0.17 mg/kg) induced region-dependent alterations in glutamate, which predicted distortions in the subjective experience of one's self (ego dissolution). Whereas higher levels of medial prefrontal cortical glutamate were associated with negatively experienced ego dissolution, lower levels in hippocampal glutamate were associated with positively experienced ego dissolution. Such findings provide further insights into the underlying neurobiological mechanisms of the psychedelic, as well as the baseline, state. Importantly, they may also provide a neurochemical basis for therapeutic effects as witnessed in ongoing clinical trials.


Low-intensity vibrations normalize adipogenesis-induced morphological and molecular changes of adult mesenchymal stem cells.
Bone marrow mesenchymal stem cells that are committed to adipogenesis were exposed daily to high-frequency low-intensity mechanical vibrations to understand molecular, morphological and ultrastructural adaptations to mechanical signals during adipogenesis. D1-ORL-UVA mouse bone marrow mesenchymal stem cells were cultured with either growth or adipogenic medium for 1 week. Low-intensity vibration signals (15 min/day, 90 Hz, 0.1 g) were applied to one group of adipogenic cells, while the other adipogenic group served as a sham control. Cellular viability, lipid accumulation, ultrastructure and morphology were determined with MTT, Oil-Red-O staining, phalloidin staining and atomic force microscopy. Semiquantitative reverse transcription polymerase chain reaction showed expression profile of the genes responsible for adipogenesis and ultrastructure of cells. Low-intensity vibration signals increased viability of the cells in adipogenic culture that was reduced significantly compared to quiescent controls. Low-intensity vibration signals also normalized the effects of adipogenic condition on cell morphology, including area, perimeter, circularization and actin cytoskeleton. Furthermore, low-intensity vibration signals reduced the expression of some adipogenic markers significantly. Mesenchymal stem cells are sensitive and responsive to mechanical loads, but debilitating conditions such as aging or obesity may steer mesenchymal stem cells toward adipogenesis. Here, daily application of low-intensity vibration signals partially neutralized the effects of adipogenic induction on mesenchymal stem cells, suggesting that these signals may provide an alternative and/or complementary option to reduce fat deposition.


Exploring the utility of phylogenetic analysis of cytochrome oxidase gene subunit I as a complementary tool to classical taxonomical identification of phlebotomine sand fly species (Diptera, Psychodidae) from southern Europe.
Phlebotomine sand flies (Diptera, Psychodidae) are known to be vectors of several pathogens such as Leishmania and Phlebovirus genera. The identification of phlebotomine sand fly species is currently based on morphological characters, and requires considerable taxonomic expertise and skilfulness, but may be complemented by DNA-based analyses for (i) accurate species identification and (ii) for estimating sand fly diversity. The aim of this study was to evaluate the utility of mitochondrial cytochrome oxidase gene subunit I (cox1) sequence analysis as a complementary tool to classical taxonomical for the identification of the most prevalent phlebotomine sand fly species from southern Europe (i.e. Phlebotomus ariasi, P. perniciosus, P. sergenti and Sergentomyia minuta). Phylogenetic analyses of cox1 sequences allowed conclusive assignment of most of the sand flies into individual species, and revealed the genetic heterogeneity that characterizes some of the identified genetic clusters. Nevertheless, it showed some limitations, as it failed to (i) allocate correctly all of all species of a given subgenus to a single lineage, or (ii) conclusively identify sequences amplified from individuals classified morphologically as P. ariasi. A more extensive analysis of cox1 sequences together with morphometric characterization of specimens from different geographic areas/regions might be useful for the correct assessment of the phylogenetic relationship within the P. ariasi/P. chadlii cluster and/or help to ascertain the usefulness of cox1 for molecular taxonomy of sand flies.


Reducing errors in prostate tracking with an improved fiducial implantation protocol for CyberKnife based stereotactic body radiotherapy (SBRT).
Ultra-hypofractionated radiotherapy with SBRT is an established technique for treating localized prostate cancer. CyberKnife based SBRT requires implantation of fiducial markers for soft tissue target tracking by the orthogonal KV X-ray imaging system. The spatial distribution of fiducial markers must allow accurate calculation of a 3D transformation that describes the position of the prostate within the reference frame of the planning CT scan. Accuray provides a fiducial implantation guideline for tracking soft tissue lesions. Despite using the guideline we experienced an unacceptably high rate of rotational tracking failure due to problems with fiducial placement. We adapted the Accuray guideline to prostate SBRT for improved fiducial placement and more reliable target tracking.Methods and materials: 54 patients with prostate adenocarcinoma were treated with ultra-hypofractionated radiotherapy on CyberKnife. Patients had platinum fiducial markers implanted transrectally under ultrasound guidance by a Radiologist. For the first 26 patients, fiducial markers were positioned following the Accuray fiducial placement guidelines for soft tissue lesions (cohort 1). The initial rotational tracking error rate was unacceptably high (23%). On review, inappropriate fiducial placement was identified as the cause of error (especially insufficient spacing between seeds). In October 2016 we developed a seed placement protocol specifically for implanting fiducial markers within the prostate and a second cohort of patients was treated thereafter (cohort 2, 28 patients). The stipulations of the original guideline are maintained while the modified protocol requires that 4 fiducial markers be implanted in the postero-lateral peripheral zone in a single coronal plane. In cohort 1, patients had a median age of 64 years (50 - 74), PSA of 6.6mcg/L (1.1 - 14.7), and prostate volume of 56 cc (22 - 125), while in cohort 2 they had a mean age of 65 years (53 - 75), PSA of 6.2 mcg/L (1 - 12) and prostate volume of 47 cc (21 - 106). The fiducial markers were easily visualized and there were no cases of urosepsis related to fiducial implantation. In 6 of 26 patients (23%) from cohort 1, only translational mapping without accurate spatial rotations could be calculated. After adopting the prostate specific fiducial implantation protocol, rotational tracking error was eliminated. Accurate 6 degree tracking (accounting for translations and rotations) was achieved in all 28 patients from cohort 2. Using an in-house computer script we analyzed the dose distributions resulting from rotational misalignments of -10, -5, -3, 3, 5, and 10 degrees along all three rotational axes (pitch, roll and yaw). Rotational misalignments result in decreased minimum dose to the PTV and increased maximum dose to OARs. Implementing a prostate specific fiducial placement protocol for SBRT significantly improved our ability to track prostate motion in 6 degrees 77% to 100% reliability. Failure to track rotations can potentially lead to underdosing and overdosing of portions of the prostate and OARs respectively.


Clonal population structure and genetic variation in sand-shinnery oak,Quercus havardii (Fagaceae).
We investigated clonal population structure and genetic variation in Quercus havardii (sand-shinnery oak), a deciduous rhizomatous shrub that dominates vegetation by forming uninterrupted expanses of ground cover over sandy deposits on the plains of western Texas, western Oklahoma, and eastern New Mexico. Isozyme electrophoresis (15 loci coding 11 enzymes) was used to recognize and map clones arrayed in a 2000-m transect (50-m sample intervals) and a 200 × 190 m grid (10-m sample intervals). Ninety-four clones were discovered, 38 in the transect and 56 in the grid, resulting in an estimated density of ∼15 clones per hectare. Clones varied greatly in size (∼100-7000 m), shape, and degree of fragmentation. The larger clones possessed massive interiors free of intergrowth by other clones, while the smaller clones varied in degree of intergrowth. The population maintained substantial levels of genetic variation (P = 60%, A = 2.5, H(exp) = 0.289) comparable to values obtained for other Quercus spp. and for other long-lived perennials. The population was outcrossing as evidenced by conformance of most loci to Hardy-Weinberg expected genotype proportions, although exceptions indicated a limited degree of population substructuring. These data indicate that despite apparent reproduction primarily through vegetative means, Q. havardii possesses conventional attributes of a sexual population.


Electrooptic polymer materials and devices for global optical interconnects.
Global optical interconnects can provide high data rate parallel communication capability through access to the internal nodes of very large scale integrated circuits. Topographic arrays of polymeric electrooptic multilayer devices such as etalons or multilayer mirrors broadcast the data stored on the surface of the chip. Differential detection of this image permits interconnection without the need for high contrast ratios. An experimental demonstration of a point-to-point interconnection using a Fabry-Perot etalon with a polymeric thin film spacer is presented.


Facile synthesis 3D flexible core-shell graphene/glass fiber via chemical vapor deposition.
Direct deposition of graphene layers on the flexible glass fiber surface to form the three-dimensional (3D) core-shell structures is offered using a two-heating reactor chemical vapor deposition system. The two-heating reactor is utilized to offer sufficient, well-proportioned floating C atoms and provide a facile way for low-temperature deposition. Graphene layers, which are controlled by changing the growth time, can be grown on the surface of wire-type glass fiber with the diameter from 30 nm to 120 um. The core-shell graphene/glass fiber deposition mechanism is proposed, suggesting that the 3D graphene films can be deposited on any proper wire-type substrates. These results open a facile way for direct and high-efficiency deposition of the transfer-free graphene layers on the low-temperature dielectric wire-type substrates. 81.05.U-; 81.07.-b; 81.15.Gh.


Microarray and proteome array in an atherosclerosis mouse model for identification of biomarkers in whole blood.
Cardiovascular disease (CVD) is highly fatal, and 80 percent of the mortality is attributed to heart attack and stroke. Atherosclerosis is a disease that increases a patient's risk to CVD and is characterized by atheroma formed by immune cells, lipids, and smooth muscle cells. When an atherosclerotic lesion grows and blocks blood vessels or when an atheroma ruptures and blocks blood vessels by embolism, sudden angina, or stroke can occur. It is therefore important to diagnose atherosclerosis early and prevent its progression to more severe disease. Although myeloperoxidase, plasma fibrinogen, cardiac troponin-I, and C-reactive protein have been considered as diagnostic markers for multiple cardiac risks, specific biomarkers for atherosclerosis have not been clearly determined yet. Particularly, reliable biomarkers for the diagnosis of atherosclerosis using whole blood are not yet available. In this study, we screened potential biomarker genes and proteins from whole blood of apolipoprotein E knockout (ApoE-/- ) mice maintained on a Western diet, by comparing them to ApoE+/+ mice. We used whole blood for microarray and proteome array. Candidate genes and proteins identified from each method were confirmed with quantitative real-time PCR and ELISA. Based on our data, we speculate that Lilrb4a, n-R5s136, and IL-5 are potential targets that can be developed into novel biomarkers of atherosclerosis. Our study contributes to the diagnosis of atherosclerosis using whole blood in clinical settings.