Heartlift patients? An interview-based study of GP trainers and the impact of 'patients they like'.
The concept of the 'heartsink patient' is well known and much used when talking about general practice. The opposite of this type of patient, however, has been little explored. To identify patient characteristics valued by GPs. Structured interview to collect narratives from GPs of individual patients, analysed qualitatively through thematic analysis and word frequency. Primary Care in Ireland. GP trainers. Emergent themes from four lead questions: Tell me about a patient you like, Tell me about the patient's personality, What have you learned about yourself as a GP?, What is different about being a GP as opposed to any other kind of doctor? In addition, a corpus linguistic analysis of word frequencies disclosed further themes, not identifiable on the surface of discourse. Ten themes were identified: GPs valued patients who were likeable, a challenge, involved them in negotiation of the doctor-patient relationship, were interesting or virtuous and had a positive effect. GPs valued their profession in that they were facilitators, gave and elicited loyalty, formed personal attachments and had a different perspective. 'Heartlift patients' may be a robust concept, to counterbalance heartsink patients. Data collected are suitable for training, and could help GPs enhance a sense of vocation.


Pathogenic Yersinia Promotes Its Survival by Creating an Acidic Fluid-Accessible Compartment on the Macrophage Surface.
Microbial pathogens and host immune cells each initiate events following their interaction in an attempt to drive the outcome to their respective advantage. Here we show that the bacterial pathogen Yersinia pseudotuberculosis sustains itself on the surface of a macrophage by forming acidic fluid-accessible compartments that are partially bounded by the host cell plasma membrane. These Yersinia-containing acidic compartments (YACs) are bereft of the early endosomal marker EEA1 and the lysosomal antigen LAMP1 and readily form on primary macrophages as well as macrophage-like cell lines. YAC formation requires the presence of the Yersinia virulence plasmid which encodes a type III secretion system. Unexpectedly, we found that the initial formation of YACs did not require translocation of the type III effectors into the host cell cytosol; however, the duration of YACs was markedly greater in infections using translocation-competent Y. pseudotuberculosis strains as well as strains expressing the effector YopJ. Furthermore, it was in this translocation- and YopJ-dependent phase of infection that the acidic environment was critical for Y. pseudotuberculosis survival during its interaction with macrophages. Our findings indicate that during its extracellular phase of infection Y. pseudotuberculosis initiates and then, by a separate mechanism, stabilizes the formation of a highly intricate structure on the surface of the macrophage that is disengaged from the endocytic pathway.


Genome annotation provides insight into carbon monoxide and hydrogen metabolism in Rubrivivax gelatinosus.
We report here the sequencing and analysis of the genome of the purple non-sulfur photosynthetic bacterium Rubrivivax gelatinosus CBS. This microbe is a model for studies of its carboxydotrophic life style under anaerobic condition, based on its ability to utilize carbon monoxide (CO) as the sole carbon substrate and water as the electron acceptor, yielding CO2 and H2 as the end products. The CO-oxidation reaction is known to be catalyzed by two enzyme complexes, the CO dehydrogenase and hydrogenase. As expected, analysis of the genome of Rx. gelatinosus CBS reveals the presence of genes encoding both enzyme complexes. The CO-oxidation reaction is CO-inducible, which is consistent with the presence of two putative CO-sensing transcription factors in its genome. Genome analysis also reveals the presence of two additional hydrogenases, an uptake hydrogenase that liberates the electrons in H2 in support of cell growth, and a regulatory hydrogenase that senses H2 and relays the signal to a two-component system that ultimately controls synthesis of the uptake hydrogenase. The genome also contains two sets of hydrogenase maturation genes which are known to assemble the catalytic metallocluster of the hydrogenase NiFe active site. Collectively, the genome sequence and analysis information reveals the blueprint of an intricate network of signal transduction pathways and its underlying regulation that enables Rx. gelatinosus CBS to thrive on CO or H2 in support of cell growth.


Delimiting cryptic morphological variation among human malaria vector species using convolutional neural networks.
Deep learning is a powerful approach for distinguishing classes of images, and there is a growing interest in applying these methods to delimit species, particularly in the identification of mosquito vectors. Visual identification of mosquito species is the foundation of mosquito-borne disease surveillance and management, but can be hindered by cryptic morphological variation in mosquito vector species complexes such as the malaria-transmitting Anopheles gambiae complex. We sought to apply Convolutional Neural Networks (CNNs) to images of mosquitoes as a proof-of-concept to determine the feasibility of automatic classification of mosquito sex, genus, species, and strains using whole-body, 2D images of mosquitoes. We introduce a library of 1, 709 images of adult mosquitoes collected from 16 colonies of mosquito vector species and strains originating from five geographic regions, with 4 cryptic species not readily distinguishable morphologically even by trained medical entomologists. We present a methodology for image processing, data augmentation, and training and validation of a CNN. Our best CNN configuration achieved high prediction accuracies of 96.96% for species identification and 98.48% for sex. Our results demonstrate that CNNs can delimit species with cryptic morphological variation, 2 strains of a single species, and specimens from a single colony stored using two different methods. We present visualizations of the CNN feature space and predictions for interpretation of our results, and we further discuss applications of our findings for future applications in malaria mosquito surveillance.


Evaluation of the duplication of staging CT scans for localized colon cancer in a Medicare population.
To quantify and characterize duplicated tests performed during the staging of localized colon cancer in the Medicare population. We used the SEER-Medicare linked database to select patients diagnosed with localized colon cancer between the years 1996 and 2009. We considered a patient as adequately staged after having received a colonoscopy, an abdominal computed tomography (CT) scan, and a pelvic CT scan. Abdominal and pelvic CT scans performed between complete staging and first cancer-directed treatment, if not ordered due to an acute condition, were considered duplicates. We characterized the institutions providing the tests and evaluated the association with survival using a weighted pooled logistic regression adjusted by baseline and time-varying confounders. Of 36,291 patients with a complete staging, 2680 (7.4%) had at least 1 duplicated test. Patients receiving a duplicate had a higher comorbidity score, were more symptomatic, and had more visits to the emergency department and clinical evaluations. They also were treated with surgery less frequently and had worse survival (hazard ratio 1.22, 95% confidence interval, 1.16-1.28). The type of institution involved in the staging (nonprofit/government centers, proprietary centers, free-standing facilities) was not associated with receiving duplicated tests. We found a low frequency of duplicated abdominal or pelvic CT scans in the staging of colon cancer in the Medicare population.


State tobacco control spending and youth smoking.
We examined the relationship between state-level tobacco control expenditures and youth smoking prevalence and cigarette consumption. We estimated a 2-part model of cigarette demand using data from the 1991 through 2000 nationally representative surveys of 8th-, 10th-, and 12th-grade students as part of the Monitoring the Future project. We found that real per capita expenditures on tobacco control had a negative and significant impact on youth smoking prevalence and on the average number of cigarettes smoked by smokers. Had states represented by the Monitoring the Future sample and the District of Columbia spent the minimum amount of money recommended by the Centers for Disease Control and Prevention, the prevalence of smoking among youths would have been between 3.3% and 13.5% lower than the rate we observed over this period.


Participatory assessment of animal health and husbandry practices in smallholder pig production systems in three high poverty districts in Uganda.
While animal health constraints have been identified as a major limiting factor in smallholder pig production in Uganda, researchers and policy makers lack information on the relative incidence of diseases and their impacts on pig production. This study aimed to assess animal health and management practices, constraints and opportunities for intervention in smallholder pig value chains in three high poverty districts of Uganda. Semi-qualitative interview checklists through Focus Group Discussions (FGDs) were administered to 340 pig farmers in 35 villages in Masaka, Kamuli and Mukono districts. Quantitative data was obtained during the exercise through group consensus. Results of FGDs were further triangulated with secondary data and information obtained from key informant interviews. Findings show that pig keeping systems are dominated by tethering and scavenging in rural areas. In peri-urban and urban areas, intensive production systems are more practiced, with pigs confined in pens. The main constraints identified by farmers include high disease burden such as African swine fever (ASF) and parasites, poor housing and feeding practices, poor veterinary services, ineffective drugs and a general lack of knowledge on piggery management. According to farmers, ASF is the primary cause of pig mortality with epidemics occurring mainly during the dry season. Worms and ectoparasites namely; mange, lice and flies are endemic leading to stunted growth which reduces the market value of pigs. Diarrhoea and malnutrition are common in piglets. Ninety-three percent of farmers say they practice deworming, 37% practice ectoparasite spraying and 77% castrate their boars. Indigenous curative treatments include the application of human urine and concoctions of local herbs for ASF control and use of old engine oil or tobacco extracts to control ectoparasites. There is a need for better technical services to assist farmers with these problems.


Coherent anti-Stokes Raman scattering microscopy for quantitative characterization of mixing and flow in microfluidics.
We present an optical, noninvasive and label-free approach to characterize flow profiles in microfluidic devices. Coherent anti-Stokes Raman scattering signals were used to map the mass transport in a microfluidic device that was then related back to the local flow rate of dilute solutes having constant fluid properties. Flow characterization was demonstrated in two common types of microfluidic devices, polydimethylsiloxane/glass square channels and wet-etched glass tapered channels.


Efficacy of selective venous catheterization in localizing a small androgen-producing tumor in ovary.
Androgen-producing tumors, originating mostly in the ovary or adrenal gland, induce hirsutism. It sometimes is difficult to localize the tumor precisely even with modern imaging technology. We used selective venous catheterization and hormonal sampling (SVCHS) to localize an androgen-producing ovarian tumor. A 37-year-old woman (gravida 0, para 0) presented with secondary amenorrhea, infertility, and virilization, including hirsutism and progressive balding. Laboratory examination revealed severe hyperandrogenism, with a total testosterone (T) concentration of 13.1 ng/ml and a free T concentration of 28.1 pg/ml. Dehydroepiandrosterone sulfate and androstendione were within normal ranges. Work-up included an abdominal and pelvic ultrasound scan, CT, MRI, and norcholesterol scintigraphy without discovery of the source of the hyperandrogenism. Persistently high plasma T concentrations prompted SVCHS. Eleven blood samples were collected from both the adrenal and the ovarian veins bilaterally. The total T concentration was significantly higher in blood from the right ovarian vein (878 ng/ml). A laparoscopic right oophorectomy was performed. The pathologic diagnosis was a Leydig cell tumor. A rapid decrease in free and total T followed tumor removal, and she became pregnant by in vitro fertilization. SVCHS is highly effective in confirming the presence of a small androgen-producing ovarian tumor.


Effective treatment of respiratory alphaherpesvirus infection using RNA interference.
Equine herpesvirus type 1 (EHV-1), a member of the Alphaherpesvirinae, is spread via nasal secretions and causes respiratory disease, neurological disorders and abortions. The virus is a significant equine pathogen, but current EHV-1 vaccines are only partially protective and effective metaphylactic and therapeutic agents are not available. Small interfering RNAs (siRNA's), delivered intranasally, could prove a valuable alternative for infection control. siRNA's against two essential EHV-1 genes, encoding the viral helicase (Ori) and glycoprotein B, were evaluated for their potential to decrease EHV-1 infection in a mouse model. METHODOLOGY/PRINCIPAL FNDINGS: siRNA therapy in vitro significantly reduced virus production and plaque size. Viral titers were reduced 80-fold with 37.5 pmol of a single siRNA or with as little as 6.25 pmol of each siRNA when used in combination. siRNA therapy in vivo significantly reduced viral replication and clinical signs. Intranasal treatment did not require a transport vehicle and proved effective when given up to 12 h before or after infection. siRNA treatment has potential for both prevention and early treatment of EHV-1 infections.


Patients with acute stroke are less likely to be admitted directly to a stroke unit when hospital beds are scarce: A Swedish multicenter register study.
It is well established that managing patients with acute stroke in dedicated stroke units is associated with improved functioning and survival. The objectives of this study are to investigate whether patients with acute stroke are less likely to be directly admitted to a stroke unit from the Emergency Department when hospital beds are scarce and to measure variation across hospitals in terms of this outcome. This register study comprised data on patients with acute stroke admitted to 14 out of 72 Swedish hospitals in 2011-2014. Data from the Swedish stroke register were linked to administrative daily data on hospital bed occupancy (measured at 6 a.m.). Logistic regression analysis was used to analyse the association between bed occupancy and direct stroke unit admission. A total of 13,955 hospital admissions were included; 79.6% were directly admitted to a stroke unit from the Emergency Department. Each percentage increase in hospital bed occupancy was associated with a 1.5% decrease in odds of direct admission to a stroke unit (odds ratio = 0.985, 95% confidence interval = 0.978-0.992). The best-performing hospital exhibited an odds ratio of 3.8 (95% confidence interval = 2.6-5.5) for direct admission to a stroke unit versus the reference hospital. We found an association between hospital crowding and reduced quality of care in acute stroke, portrayed by a lower likelihood of patients being directly admitted to a stroke unit from the Emergency Department. The magnitude of the effect varied considerably across hospitals.


Oligomycins as inhibitors of K-Ras plasma membrane localisation.
Frequently present in pancreatic, colorectal and non-small cell lung carcinomas, oncogenic mutant K-Ras must be localised to the plasma membrane (PM) to be functional. Inhibitors of K-Ras PM localisation are therefore putative cancer chemotherapeutics. By screening a microbial extract library in a high content cell-based assay we detected the rare oligomycin class of Streptomyces polyketides as inhibitors of K-Ras PM localisation. Cultivation and fractionation of three unique oligomycin producing Streptomyces strains yielded oligomycins A-E (1-5) and 21-hydroxy-oligomycin A (6), together with the new 21-hydroxy-oligomycin C (7) and 40-hydroxy-oligomycin B (8). Structures for 1-8 were assigned by detailed spectroscopic analysis. Cancer cell viability screening confirmed 1-8 were cytotoxic to human colorectal carcinoma cells (IC50 > 3 μM), and were inhibitors of the ABC transporter efflux pump P-glycoprotein (P-gp), with 5 being comparable in potency to the positive control verapamil. Significantly, oligomycins 1-8 proved to be exceptionally potent inhibitors of K-Ras PM localisation (Emax 0.67-0.75 with an IC50 ~ 1.5-14 nM).


Developmental mechanisms underlying improved contrast thresholds for discriminations of orientation signals embedded in noise.
We combined an external noise paradigm with an efficient procedure for obtaining contrast thresholds (Lesmes et al., 2006) in order to model developmental changes in the effect of noise on contrast discrimination during childhood. Specifically, we measured the contrast thresholds of 5-, 7-, 9-year-olds and adults (n = 20/age) in a two alternative forced-choice orientation discrimination task over a wide range of external noise levels and at three levels of accuracy. Overall, as age increased, contrast thresholds decreased over the entire range of external noise levels tested. The decrease was greatest between 5 and 7 years of age. The reduction in threshold after age 5 was greater in the high than the low external noise region, a pattern implying greater tolerance of the irrelevant background noise as children became older. To model the mechanisms underlying these developmental changes in terms of internal noise components, we adapted the original perceptual template model (Lu and Dosher, 1998) and normalized the magnitude of performance changes against the performance of 5-year-olds. The resulting model provided an excellent fit (r (2) = 0.985) to the contrast thresholds at multiple levels of accuracy (60, 75, and 90%) across a wide range of external noise levels. The improvements in contrast thresholds with age were best modeled by a combination of reductions in internal additive noise, reductions in internal multiplicative noise, and improvements in excluding external noise by template retuning. In line with the data, the improvement was greatest between 5 and 7 years of age, accompanied by a 39% reduction in additive noise, 71% reduction in multiplicative noise, and 45% improvement in external noise exclusion. The modeled improvements likely reflect developmental changes at the cortical level, rather than changes in front-end structural properties (Kiorpes et al., 2003).


Review of influenza A virus in swine worldwide: a call for increased surveillance and research.
Pigs and humans have shared influenza A viruses (IAV) since at least 1918, and many interspecies transmission events have been documented since that time. However, despite this interplay, relatively little is known regarding IAV circulating in swine around the world compared with the avian and human knowledge base. This gap in knowledge impedes our understanding of how viruses adapted to swine or man impacts the ecology and evolution of IAV as a whole and the true impact of swine IAV on human health. The pandemic H1N1 that emerged in 2009 underscored the need for greater surveillance and sharing of data on IAV in swine. In this paper, we review the current state of IAV in swine around the world, highlight the collaboration between international organizations and a network of laboratories engaged in human and animal IAV surveillance and research, and emphasize the need to increase information in high-priority regions. The need for global integration and rapid sharing of data and resources to fight IAV in swine and other animal species is apparent, but this effort requires grassroots support from governments, practicing veterinarians and the swine industry and, ultimately, requires significant increases in funding and infrastructure.


Influence of abomasal carbohydrates on subcutaneous, omental, and mesenteric adipose lipogenic and lipolytic rates in growing beef steers.
To determine the response to alteration in site and form of carbohydrate delivery to the digestive tract, in vitro rates of lipogenesis and lipolysis in mesenteric (MESA), omental (OMA), and subcutaneous (SQA) adipose depots were compared. Forty crossbred beef steers (243 +/- 2 kg of BW) were fed 161 (LI) or 214 (HI) kcal of ME/(kg of BW(0.75) x d) or they were fed LI and infused for 35 d into the rumen (R) or abomasum (A) with starch hydrolysate (SH) or into the abomasum with glucose (G). Jugular blood samples were collected, steers were slaughtered, and adipose depots were sampled and prepared for assessment of lipogenesis and lipolysis in vitro. Blood concentrations of glucagon were increased (P = 0.04) in HI-H2O compared with LI-H2O steers, whereas A-SH tended to increase (P = 0.08) circulating IGF-I relative to R-SH, and A-G tended to have elevated (P = 0.09) T3 compared with A-SH. Lipolysis, as assessed by NEFA release, was unaffected by treatment. Glycerol release by the MESA and SQA was increased or tended to be increased (P < or = 0.08) in HI-H2O compared with LI-H2O steers. In A-G compared with A-SH steers, glycerol release from OMA increased (P = 0.008) and from SQA tended to be increased (P = 0.08). Acetate incorporation into total neutral lipids (TNL) increased or tended to increase with ME intake and SH infusion (P < or = 0.09) across all depots. Rates of acetate incorporation into fatty acids (FA) also increased or tended to be increased (P < or = 0.1) by SH infusion across all depots, but only that of SQA was increased with ME intake (HI-H2O vs. LI-H2O; P = 0.02). Rates of acetate incorporation into FA and TNL in MESA were increased (P < or = 0.03) by A-SH compared with R-SH, but site of SH infusion did not affect the rates in SQA or OMA. Glucose incorporation into TNL for MESA and SQA increased or tended to be increased (P < or = 0.1) by dietary and infused energy, whereas for OMA they tended to be increased (P = 0.1) only by SH infusion. In contrast, glucose incorporation into FA was unaffected by energy supply but tended to be increased (P = 0.07) by SH in MESA and tended to be greater (P = 0.08) for A-G than A-SH in OMA. The general across-depot pattern of acetate incorporation rate into FA and TNL was SQA > OMA > MESA, whereas, for glucose incorporation, rates across depots were equivalent. These data provide evidence that the postruminal supply of energy, specifically carbohydrate, stimulates lipogenesis from acetate and glucose and is more pronounced in abdominal depots relative to the subcutaneous depot.


Sustainability. Planetary boundaries: guiding human development on a changing planet.
The planetary boundaries framework defines a safe operating space for humanity based on the intrinsic biophysical processes that regulate the stability of the Earth system. Here, we revise and update the planetary boundary framework, with a focus on the underpinning biophysical science, based on targeted input from expert research communities and on more general scientific advances over the past 5 years. Several of the boundaries now have a two-tier approach, reflecting the importance of cross-scale interactions and the regional-level heterogeneity of the processes that underpin the boundaries. Two core boundaries—climate change and biosphere integrity—have been identified, each of which has the potential on its own to drive the Earth system into a new state should they be substantially and persistently transgressed.


Association between quantitative high-risk human papillomavirus DNA load and cervical intraepithelial neoplasm risk.
Human papillomavirus (HPV) infection is a high-risk factor for cervical intraepithelial neoplasm (CIN) but the association between the quantitative HPV DNA load and the severity of CIN remains controversial. We conducted a community study to investigate the correlation between the two. Potential study subjects were selected through Pap smear screening in Kaohsiung County, Taiwan. Ninety-one subjects with either their first case of inflammation or > or =CIN1 by biopsy confirmation were assigned to a case group; 175 normal subjects with negative findings by Pap smears or biopsies were assigned to a control group. Cervical HPV load was detected with Hybrid Capture II assay for high-risk HPV infection, with nested PCR for high- and low-risk HPV infection, and with type-specific PCR for HPV type 16 (HPV-16). Individuals with positive high-risk HPV infection had an increased risk of developing CIN. Compared with HPV-negative subjects, the odds ratios were 32.2 [95% confidence interval (95% CI), 10.4-99.5] for subjects with CIN1, 37.2 (95% CI, 7.4-187.6) for subjects with CIN2, and 68.3 (95% CI, 14.1-328.5) for subjects with > or =CIN3 after adjusting for other confounding factors. The similar trend was also found among the HPV-16-negative individuals. In addition, high-risk HPV DNA load levels were highly correlated with the different grades of CINs in the overall population (Spearman's correlation coefficient r = 0.67, P < 0.0001, n = 266) and the HPV-16-negative population (Spearman's correlation coefficient r = 0.58, P < 0.0001, n = 234). We concluded that high-risk HPV infection, irrespective of HPV-16 infection, was highly and positively associated with the development of CIN.


The clinical and integrated management of COPD. An official document of AIMAR (Interdisciplinary Association for Research in Lung Disease), AIPO (Italian Association of Hospital Pulmonologists), SIMER (Italian Society of Respiratory Medicine), SIMG (Italian Society of General Medicine).
COPD is a chronic pathological condition of the respiratory system characterized by persistent and partially reversible airflow obstruction, to which variably contribute remodeling of bronchi (chronic bronchitis), bronchioles (small airway disease) and lung parenchyma (pulmonary emphysema). COPD can cause important systemic effects and be associated with complications and comorbidities. The diagnosis of COPD is based on the presence of respiratory symptoms and/or a history of exposure to risk factors, and the demonstration of airflow obstruction by spirometry. GARD of WHO has defined COPD "a preventable and treatable disease". The integration among general practitioner, chest physician as well as other specialists, whenever required, assures the best management of the COPD person, when specific targets to be achieved are well defined in a diagnostic and therapeutic route, previously designed and shared with appropriateness. The first-line pharmacologic treatment of COPD is represented by inhaled long-acting bronchodilators. In symptomatic patients, with pre-bronchodilator FEV1 < 60% predicted and ≥ 2 exacerbations/year, ICS may be added to LABA. The use of fixed-dose, single-inhaler combination may improve the adherence to treatment. Long term oxygen therapy (LTOT) is indicated in stable patients, at rest while receiving the best possible treatment, and exhibiting a PaO2 ≤ 55 mmHg (SO2 < 88%) or PaO2 values between 56 and 59 mmHg (SO2 < 89%) associated with pulmonary arterial hypertension, cor pulmonale, or edema of the lower limbs or hematocrit > 55%. Respiratory rehabilitation is addressed to patients with chronic respiratory disease in all stages of severity who report symptoms and limitation of their daily activity. It must be integrated in an individual patient tailored treatment as it improves dyspnea, exercise performance, and quality of life. Acute exacerbation of COPD is a sudden worsening of usual symptoms in a person with COPD, over and beyond normal daily variability that requires treatment modification. The pharmacologic therapy can be applied at home and includes the administration of drugs used during the stable phase by increasing the dose or modifying the route, and adding, whenever required, drugs as antibiotics or systemic corticosteroids. In case of patients who because of COPD severity and/or of exacerbations do not respond promptly to treatment at home hospital admission should be considered. Patients with "severe" or "very severe" COPD who experience exacerbations should be carried out in respiratory unit, based on the severity of acute respiratory failure. An integrated system is required in the community in order to ensure adequate treatments also outside acute care hospital settings and rehabilitation centers. This article is being simultaneously published in Sarcoidosis Vasc Diffuse Lung Dis 2014, 31(Suppl. 1);3-21.


Attention-deficit/hyperactivity disorder subtypes in adolescents with comorbid substance-use disorder.
Little is known about the relationship between attention-deficit/hyperactivity disorder (ADHD) subtypes and substance-use disorder (SUD). As there is literature suggesting different subtype phenotypes, there may be subtype differences in regard to the risk for developing SUD and substance treatment response. To characterize the sample in a Clinical Trials Network (CTN) study according to ADHD subtypes and baseline psychosocial and substance-use characteristics and to compare subtypes on response to treatment. Secondary analyses on data collected from adolescents (n = 276) diagnosed with ADHD and SUD (non-nicotine) and treated with stimulant medication or placebo and cognitive behavioral therapy (CBT) for substance use. Participants were characterized as inattentive or combined ADHD subtype and compared on baseline characteristics and treatment outcome. The combined subtype presented with more severe SUDs and higher rates of conduct disorder. There were a greater proportion of boys with inattentive subtype. The inattentive subtype appeared less ready for treatment (greater University of Rhode Island Change Assessment precontemplation scores) with poorer coping skills (poorer problem-solving and abstinence focused coping) at baseline. However, the two subtypes responded equally to treatment even after controlling for baseline differences. Findings from this large community sample indicate that there were no subtype differences in treatment response, although there were differences in terms of substance use, antisocial behavior, readiness for treatment, and gender prior to treatment. This study is the first to report on subtype differences for treatment response for non-nicotine SUD in a comorbid ADHD-SUD population. Despite some baseline differences, both subtypes responded equally to treatment, suggesting limited relevance for subtype designation on treatment planning.


Alarm calls elicit predator-specific physiological responses.
Glucocorticoids regulate glucose concentrations and responses to unpredictable events, while also modulating cognition. Juvenile Belding's ground squirrels (Urocitellus beldingi) learn to respond to whistle and trill alarm calls, warning of aerial and terrestrial predators, respectively, shortly after emerging from natal burrows at one month of age. Alarm calls can cause physiological reactions and arousal, and this arousal, coupled with watching adult responses, might help juveniles learn associations between calls and behavioural responses. I studied whether young show differential cortisol responses to alarm and non-alarm calls, using playbacks of U. beldingi whistles, trills, squeals (a conspecific control vocalization) and silent controls. Trills elicited very high cortisol responses, and, using an individual's response to the silent control as baseline, only their response to a trill was significantly higher than baseline. This cortisol increase would provide glucose for extended vigilance and escape efforts, which is appropriate for evading terrestrial predators which hunt for long periods. Although whistles do not elicit a cortisol response, previous research has shown that they do result in bradycardia, which enhances attention and information processing. This is a novel demonstration of two physiological responses to two alarm calls, each appropriate to the threats represented by the calls.


Shear-Mediated Platelet Activation Enhances Thrombotic Complications in Patients With LVADs and Is Reversed After Heart Transplantation.
We present a time-series analysis of shear-mediated platelet activation (SMPA) in a patient supported by a Left Ventricular Assist Device (LVAD) who developed recurrent thromboembolic events over the course of support. The patient showed marked alterations of SMPA as quantified via the Platelet Activity State (PAS) assay. At the time of a first pump thrombosis, PAS was equal to 16.2 ± 2.1%, indicating a significant level of SMPA. A moderate decrease of PAS was observed 30 days after pump exchange (PAS = 9.5 ± 1.0%). A sudden PAS increase was recorded after 60 days (PAS = 60.5 ± 4.3%), and an ischemic stroke with hemorrhagic transformation was subsequently diagnosed at 120 days after pump exchange (PAS = 86.9 ± 7.5%); high PAS values did not recover over time likely because of sustained platelet activation from the inflammatory milieu caused by a driveline infection occurred at 240 days of support (PAS = 84.4 ± 7.3%). PAS values were completely reversed after heart transplantation (PAS = 0.6 ± 0.1%), demonstrating resolution of SMPA. This study provides further insight into our understanding of the pathogenesis of LVAD thrombosis, addressing SMPA as a relevant key factor associated with thrombotic complications. With the PAS assay, we have identified a reliable biomarker to promote tailored pharmacological therapy for the prevention of thromboembolic events in patients with LVADs.


Deletion of the GluA1 AMPA receptor subunit impairs recency-dependent object recognition memory.
Deletion of the GluA1 AMPA receptor subunit impairs short-term spatial recognition memory. It has been suggested that short-term recognition depends upon memory caused by the recent presentation of a stimulus that is independent of contextual-retrieval processes. The aim of the present set of experiments was to test whether the role of GluA1 extends to nonspatial recognition memory. Wild-type and GluA1 knockout mice were tested on the standard object recognition task and a context-independent recognition task that required recency-dependent memory. In a first set of experiments it was found that GluA1 deletion failed to impair performance on either of the object recognition or recency-dependent tasks. However, GluA1 knockout mice displayed increased levels of exploration of the objects in both the sample and test phases compared to controls. In contrast, when the time that GluA1 knockout mice spent exploring the objects was yoked to control mice during the sample phase, it was found that GluA1 deletion now impaired performance on both the object recognition and the recency-dependent tasks. GluA1 deletion failed to impair performance on a context-dependent recognition task regardless of whether object exposure in knockout mice was yoked to controls or not. These results demonstrate that GluA1 is necessary for nonspatial as well as spatial recognition memory and plays an important role in recency-dependent memory processes.


Dual-Level Capacitive Micromachined Uncooled Thermal Detector.
This paper presents a novel dual-level capacitive microcantilever-based thermal detector that is implemented in the commercial surface micromachined PolyMUMPs technology. The proposed design is implemented side-by-side with four different single-level designs to enable a design-to-design performance comparison. The dual-level design exhibits a rate of capacitance change per degree Celsius that is over three times higher than that of the single-level designs and has a base capacitance that is more than twice as large. These improvements are achieved because the dual-level architecture allows a 100% electrode-to-detector area, while single-level designs are shown to suffer from an inherent trade-off between sensitivity and base capacitance. In single-level designs, either the number of the bimorph beams or the capacitance electrode can be increased for a given sensor area. The former is needed for a longer effective length of the bimorph for higher thermomechanical sensitivity (i.e., larger tilting angels per degree Celsius), while the latter is desired to relax the read-out integrated-circuits requirements. This thermomechanical response-to-initial capacitance trade-off is mitigated by the dual-level design, which dedicates one structural layer to serve as the upper electrode of the detector, while the other layer contains as many bimorph beams as desired, independently of the former's area.


Fluoride Content in Alcoholic Drinks.
The aim of the study was to determine the role of alcoholic drinks as a potential source of dietary fluoride by means of measuring fluoride levels in selected alcoholic drinks available on the Polish market that are also diverse in terms of the percentage content of ethanol. The study was conducted on 48 types of drinks with low, medium, and high alcohol content available on the Polish market and offered by various manufacturers, both Polish and foreign. Fluoride concentrations in individual samples were measured by potentiometric method with a fluoride ion-selective electrode. The highest fluoride levels were determined in the lowest percentage drinks (less than 10 % v/v ethanol), with the lowest fluoride levels observed in the highest percentage drinks (above 40 % v/v ethanol). In terms of types of alcoholic drinks, the highest fluoride levels were determined in beers and wines, while the lowest levels were observed in vodkas. These data confirm the fact that alcoholic beverages need to be considered as a significant source of fluoride delivered into the body.


A novel evolutionarily conserved element is a general transcriptional repressor of p21WAF¹/CIP¹.
The effective induction of p21(WAF1/CIP1/Cdkn1a) (p21) expression in p53-negative cancer cells is an important avenue in cancer management. We investigated the ability of various common chemotherapeutic drugs to induce p21 expression in p53-negative cancer cells and showed that the induction of p21 expression by oxaliplatin is caused by the derepression of a previously unrecognized negative regulatory element with a Sp1/Sp3 palindrome sequence core at -216 to -236 of the p21 proximal promoter. Electrophoretic mobility shift and antibody super-shift assays confirmed the specific binding of Sp1/Sp3, and showed that the oxaliplatin-mediated derepression of p21 transcription was associated with an increased Sp1/Sp3 phosphorylation and binding affinity to the oxaliplatin-responsive element. A search of the ENCODE database for vertebrate-conserved genomic elements identified the Sp1/Sp3 palindrome element as the only vertebrate-conserved element within the 500-bp proximal p21 promoter region, indicating its fundamental importance. In in vivo competition assays, transfected synthetic Sp1/Sp3 palindrome elements derepressed the cotransfected or endogenous p21 promoter in a dosage-dependent manner. This derepression was not seen in oxaliplatin-treated cells, suggesting that the exogenous Sp1/Sp3 palindrome and oxaliplatin had the same downstream signaling target. Taken together, our results revealed, for the first time, this evolutionarily conserved Sp1/Sp3 palindrome element in the proximal p21 promoter that serves as a regulatory repressor to maintain p21 basal level expression.


Clinical and epidemiological characterisation of lymphogranuloma venereum in southwest Spain, 2013-2015.
Lymphogranuloma venereum (LGV) infections caused by Chlamydia trachomatis L serovars have emerged in 2003 in Europe among HIV-positive men having sex with men (MSM). Our aim was to evaluate LGV prevalence and predictors in a high-risk population attending to two STI clinics in the southwest of Spain between December 2013 and April 2015. Screening of C. trachomatis using commercial kits was carried out, followed by real-time pmpH-PCR discriminating LGV strains, and finally ompA gene was sequenced for phylogenetic reconstruction. A total of 6398 samples were tested, of which, 594 (9.3%) were C. trachomatis-positive specimens and successfully typed by pmpH PCR. Five hundred and eighty-one samples contained non-LGV and 13 (2.2%; 95% CI 1.3% to 3.7%) samples had LGV. One hundred and sixty-six (27.9%; 95% CI 24.5% to 31.7%) CT-positive results were found in MSM. All C. trachomatis LGV types were found in rectal samples from MSM (13/166, 7.8%; 95% CI 4.5% to 13.0%). Of these, five (38.5%; 95% CI 17.7% to 64.5%) patients were asymptomatic and 11 (84.6%; 95% CI 57.8% to 95.7%; p<0.001) were also HIV positive. Successful treatment of LGV was achieved in all patients including 11/13 (84.6%) who received single-dose azithromycin. All of the L types were confirmed to be genotype L2b with ompA PCR and sequencing. This analysis shows that LGV infections are occurring in MSM in southwest Spain, where no data about LGV have been described before, reinforcing the need for screening and genotyping for LGV. LGV should be taken into account when considering treatment and management of rectal C. trachomatis infections, including in asymptomatic HIV-positive MSM. Larger studies on appropriate treatment for asymptomatic LGV infection are needed.


Pandemic (H1N1) 2009 influenza vaccine uptake in pregnant women entering the 2010 influenza season in Western Australia.
To audit the uptake of pandemic (H1N1) 2009 influenza vaccine in pregnant women entering the 2010 influenza season in Western Australia, and to identify why some women did not receive the vaccine. Cross-sectional study of consecutive patients attending the Joondalup Health Campus public antenatal clinics in WA in January 2010. Audit of uptake of the H1N1-specific vaccine. Rate of H1N1-specific vaccination, and reasons for not being the vaccinated. 479 of 541 women who attended the clinics (88.5%) were included in the audit. Three women had been infected with pandemic influenza in the preceding influenza season, leaving 476 women who were eligible for vaccination in pregnancy. Of these 476 women, only 33(6.9%) had been vaccinated. Of the remaining 443 women who were eligible to receive the vaccine but had not been vaccinated, 63.9% had not been offered vaccination despite multiple visits to their general practitioners during pregnancy, 19.6% had been advised by their GPs against vaccination in pregnancy, and 61.6% stated that they would decline vaccination if offered because of safety concerns. Uptake of H1N1-specific influenza vaccine in pregnant women was poor. Reasons for this relate both to vaccination not being offered to or actively sought by the women, as well as concerns - of both the women and their GPs - about vaccine safety in pregnancy. Uptake in this setting may improve if vaccination is offered through public antenatal clinics with concurrent safety education for obstetricians and vaccination providers.


A characterization of the oral microbiome in allogeneic stem cell transplant patients.
The mouth is a complex biological structure inhabited by diverse bacterial communities. The purpose of this study is to describe the effects of allogeneic stem cell transplantation on the oral microbiota and to examine differences among those patients who acquired respiratory complications after transplantation. All patients were consented at the National Institutes of Health, Clinical Center. Bacterial DNA was analyzed from patients' oral specimens using the Human Oral Microbe Identification Microarray. The specimens were collected from four oral sites in 45 allogeneic transplantation patients. Specimens were collected at baseline prior to transplantation, after transplantation at the nadir of the neutrophil count and after myeloid engraftment. If respiratory signs and symptoms developed, additional specimens were obtained. Patients were followed for 100 days post transplantation. Eleven patients' specimens were subjected to further statistical analysis. Many common bacterial genera, such as Streptococcus, Veillonella, Gemella, Granulicatella and Camplyobacter were identified as being present before and after transplantation. Five of 11 patients developed respiratory complications following transplantation and there was preliminary evidence that the oral microbiome changed in their oral specimens. Cluster analysis and principal component analysis revealed this change in the oral microbiota. After allogeneic transplantation, the oral bacterial community's response to a new immune system was not apparent and many of the most common core oral taxa remained unaffected. However, the oral microbiome was affected in patients who developed respiratory signs and symptoms after transplantation. The association related to the change in the oral microbiota and respiratory complications after transplantation will be validated by future studies using high throughput molecular methods.


Trained-feature-specific offline learning by sleep in an orientation detection task.
Training-induced performance gains in a visual perceptual learning (VPL) task that take place during sleep are termed "offline performance gains." Offline performance gains of VPL so far have been reported in the texture discrimination task and other discrimination tasks. This raises the question as to whether offline performance gains on VPL occur exclusively in discrimination tasks. The present study examined whether offline performance gains occur in detection tasks. In Experiment 1, subjects were trained on a Gabor orientation detection task. They were retested after a 12-hr interval, which included either nightly sleep or only wakefulness. Offline performance gains occurred only after sleep on the trained orientation, not on an untrained orientation. In Experiment 2, we tested whether offline performance gains in the detection task occur over a nap using polysomnography. Moreover, we tested whether sigma activity during non-rapid eye movement (NREM) sleep recorded from occipital electrodes, previously implicated in offline performance gains of the texture discrimination task, was associated with the degree of offline performance gains of the Gabor orientation detection task. We replicated offline performance gains on the trained orientation in the detection task over the nap. Sigma activity during NREM sleep was significantly larger in the occipital electrodes relative to control electrodes in correlation with offline performance gains. The results suggest that offline performance gains occur over the sleep period generally in VPL. Moreover, sigma activity in the occipital region during NREM sleep may play an important role in offline performance gains of VPL.


Association between night/after-hours surgery and mortality: a systematic review and meta-analysis.
The association between night/after-hours surgery and patients' mortality is unclear. The protocol of this systematic review was registered in PROSPERO (CRD42019128534). We searched Medline, PubMed, and EMBASE from inception until August 29, 2019 for studies examining an association between timing of surgical procedures (time of anaesthesia induction or surgery start) and mortality (within 30 days or in-hospital) in adult patients. Studies reporting patients' mortality after surgery performed during the weekend only were excluded. All analyses were done using the random-effects model. We included 40 observational studies (36 retrospective and four prospective) that examined a total of 2 957 065 patients. Twenty-eight studies were judged of good quality and 12 of poor quality according to Newcastle-Ottawa score, owing to a lack of adequate comparability between study groups. Primary analysis from adjusted estimates demonstrated as association between night/after-hours surgery and a higher risk of mortality (odds ratio [OR]=1.16; 95% confidence interval [CI], 1.06-1.28; P=0.002; number of studies=18; I2=67%) based on low certainty evidence. Analysis from unadjusted estimates demonstrated a consistent association (OR=1.47; 95% CI, 1.19-1.83; P=0.0005; studies=38, I2=97%; low certainty). The number of centres per study had no credible subgroup effect on the association between the time of surgery and mortality. We were unable to evaluate the subgroup effect of urgency of surgery because of high heterogeneity. Night/after-hours surgery may be associated with a higher risk of mortality. Patients' and surgical characteristics seem not to completely explain this finding. However, the certainty of the evidence was low.


Testosterone control of nucleic acid content and proliferation of epithelium and stroma in rat seminal vesicles.
Tissue wet weight, nucleic acid content and epithelial and stromal cell numbers were measured in the seminal vesicles of sexually mature male rats. After castration, tissue weight and RNA decreased rapidly and in aprallel to reach, after 14 days, values only 15-20% of those in control (not castrated) animals. During this period, DNA decreased to a much lesser extent (by about 40%), but this change in DNA correlates well with the observed loss of cells from the epithelium. Testosterone in vivo promoted an immediate resynthesis of RNA, the value characteristic of control animals being reached within 80h. Delays occurred in the hormone-induced regain of tissue weight (30h) and DNA (40h), each of which preceded proliferation of the epithelium (40--50h). The cells of the stroma were unaffected by these changes in the androgenic statls of the animal. It is suggested that these proliferative changes in the epithelium cannot account for the previously reported induction by testosterone of basic secretory proteins in this tissue.


Parental attitudes toward obesity and overweight screening and communication for hospitalized children.
Many studies have evaluated BMI screening, communication, and follow-up recommendations in the outpatient setting. However, few studies have examined parental attitudes toward using the inpatient setting as a time to screen and counsel families regarding their child's BMI. We sought to study parental attitudes about overweight and obesity screening in the inpatient setting. Parents (N= 101) of children aged 2 to 18 years admitted to a general pediatric hospital or surgical service were queried regarding their attitudes about screening and counseling for overweight and obesity. Children's age, gender, height, weight, and diagnosis codes were extracted from electronic medical records and billing databases. BMI was calculated, plotted, and categorized according to standard Centers for Disease Control and Prevention growth charts and expert recommendation. Fourteen percent of children in the study were overweight, and 17% were obese. Parents of overweight and obese children underestimated their child's weight status 68% of the time. The majority believed admitted children should always have their BMI calculated. Almost all parents (90%) indicated that their inpatient physician should inform them if their child were overweight or obese and that primary care providers should be informed of the results of BMI screening. Parents of children admitted to the hospital believed their children should have their BMI screened. If their child was overweight or obese, parents believed they should be informed, and counseling should be initiated. These findings support using the inpatient time to screen and communicate BMI.


The putative phosphate transporter PitB (PP1373) is involved in tellurite uptake in Pseudomonas putida KT2440.
Tellurium oxyanions are chemical species of great toxicity and their presence in the environment has increased because of mining industries and photovoltaic and electronic waste. Recovery strategies for this metalloid that are based on micro-organisms are of interest, but further studies of the transport systems and enzymes responsible for implementing tellurium transformations are required because many mechanisms remain unknown. Here, we investigated the involvement in tellurite uptake of the putative phosphate transporter PitB (PP1373) in soil bacterium Pseudomonas putida KT2440. For this purpose, through a method based on the CRISPR/Cas9 system, we generated a strain deficient in the pitB gene and characterized its phenotype on exposing it to varied concentrations of tellurite. Growth curves and transmission electronic microscopy experiments for the wild-type and ΔpitB strains showed that both were able to internalize tellurite into the cytoplasm and reduce the oxyanion to black nano-sized and rod-shaped tellurium particles, although the ΔpitB strain showed an increased resistance to the tellurite toxic effects. At a concentration of 100 μM tellurite, where the biomass formation of the wild-type strain decreased by half, we observed a greater ability of ΔpitB to reduce this oxyanion with respect to the wild-type strain (~38 vs ~16 %), which is related to the greater biomass production of ΔpitB and not to a greater consumption of tellurite per cell. The phenotype of the mutant was restored on over-expressing pitB in trans. In summary, our results indicate that PitB is one of several transporters responsible for tellurite uptake in P. putida KT2440.


The impact of South Africa's new constitution on the organization of health services in the post-apartheid era.
This essay evaluates the transformation of the health care system in South Africa in the five years following the end of apartheid. This period has witnessed dramatic and wide-ranging changes in the roles of the central and provincial governments as well as many of the same problems related to decentralization as have been seen elsewhere in the world. The importance of effective intergovernmental relations to the creation of a national health system is highlighted. Negotiation. contracting, and coordination skills and mechanisms need to be strengthened to ensure a more effective national health system within a decentralized political system.


Homeopathic Preparations of Quartz, Sulfur and Copper Sulfate Assessed by UV-Spectroscopy.
Homeopathic preparations are used in homeopathy and anthroposophic medicine. Although there is evidence of effectiveness in several clinical studies, including double-blinded randomized controlled trials, their nature and mode of action could not be explained with current scientific approaches yet. Several physical methods have already been applied to investigate homeopathic preparations but it is yet unclear which methods are best suited to identify characteristic physicochemical properties of homeopathic preparations. The aim of this study was to investigate homeopathic preparations with UV-spectroscopy. In a blinded, randomized, controlled experiment homeopathic preparations of copper sulfate (CuSO(4); 11c-30c), quartz (SiO(2); 10c-30c, i.e., centesimal dilution steps) and sulfur (S; 11×-30×, i.e., decimal dilution steps) and controls (one-time succussed diluent) were investigated using UV-spectroscopy and tested for contamination by inductively coupled plasma mass spectrometry (ICP-MS). The UV transmission for homeopathic preparations of CuSO(4) preparations was significantly lower than in controls. The transmission seemed to be also lower for both SiO(2) and S, but not significant. The mean effect size (95% confidence interval) was similar for the homeopathic preparations: CuSO(4) (pooled data) 0.0544% (0.0260-0.0827%), SiO(2) 0.0323% (-0.0064% to 0.0710%) and S 0.0281% (-0.0520% to 0.1082%). UV transmission values of homeopathic preparations had a significantly higher variability compared to controls. In none of the samples the concentration of any element analyzed by ICP-MS exceeded 100 ppb. Lower transmission of UV light may indicate that homeopathic preparations are less structured or more dynamic than their succussed pure solvent.


The chronically disturbed adolescent and the surrogate family model: observations on two patient populations.
Two preliminary evaluations were conducted by surveying the charts of 15 chronically disturbed adolescents and young adult patients (ages 15 to 30 years) and 23 chronically disturbed adolescents (ages 12 to 18 years) treated in a tertiary surrogate family model hospital. Drug and alcohol use of patients and their parents was assessed in the first patient population. Results indicate that 40% of the patients had at least one parent who formerly used or is still addicted to drugs or alcohol (or both). The second survey examined the Global Assessment Functioning (GAF) scale administered to 23 adolescents on admission and at discharge from the hospital. Scores on the GAF had improved at discharge in 19 of 21 patients whose length of stay ranged from 2 to 12 months. The information gathered from the retrospective surveys of the medical charts of these two patient populations helps to better understand multigenerational influence as a probable factor in the development of the chronically disturbed adolescent. Also, it shows how small psychiatric hospitals may conduct ongoing outcome studies that could be useful to improve staff morale and produce evidence to case managers and third-party payers that their treatment programs provide both positive patient care and positive results.


Capturing the Dynamics of a Hybrid Multiscale Cancer Model with a Continuum Model.
Cancer is a complex disease involving processes at spatial scales from subcellular, like cell signalling, to tissue scale, such as vascular network formation. A number of multiscale models have been developed to study the dynamics that emerge from the coupling between the intracellular, cellular and tissue scales. Here, we develop a continuum partial differential equation model to capture the dynamics of a particular multiscale model (a hybrid cellular automaton with discrete cells, diffusible factors and an explicit vascular network). The purpose is to test under which circumstances such a continuum model gives equivalent predictions to the original multiscale model, in the knowledge that the system details are known, and differences in model results can be explained in terms of model features (rather than unknown experimental confounding factors). The continuum model qualitatively replicates the dynamics from the multiscale model, with certain discrepancies observed owing to the differences in the modelling of certain processes. The continuum model admits travelling wave solutions for normal tissue growth and tumour invasion, with similar behaviour observed in the multiscale model. However, the continuum model enables us to analyse the spatially homogeneous steady states of the system, and hence to analyse these waves in more detail. We show that the tumour microenvironmental effects from the multiscale model mean that tumour invasion exhibits a so-called pushed wave when the carrying capacity for tumour cell proliferation is less than the total cell density at the tumour wave front. These pushed waves of tumour invasion propagate by triggering apoptosis of normal cells at the wave front. Otherwise, numerical evidence suggests that the wave speed can be predicted from linear analysis about the normal tissue steady state.


Substrate-mediated DNA delivery: role of the cationic polymer structure and extent of modification.
DNA complex immobilization to substrates that support cell adhesion can enhance gene transfer by maintaining DNA within the cellular environment while limiting complex aggregation. This report examines the tether design (e.g., extent of functionalization) and cationic polymer structure for their effect on complex binding to the substrate and cellular transfection. DNA is complexed with cationic polymers (polylysine, PL; polyethylenimine, PEI), which are functionalized with biotin for binding to a neutravidin (NA) substrate. Surfaces densities ranging from 0.4 to 2.6 microg DNA/cm(2) were obtained for PL, and from 0.7 to 1.0 microg DNA/cm(2) for PEI. The distribution of biotin groups for PL/DNA complexes had a dual effect on cellular transfection. Increasing the fraction of PL with biotin residues decreased luciferase activity; however, increasing the number of biotin residues per PL increased luciferase activity. For PEI, the number of biotin groups present on the complex did not affect transgene expression. Release studies demonstrated that 20-30% of the immobilized DNA was released over 8 days, with 8-20% released during the first 24 h. Enzymatic degradation of cationic polymers is not necessary for transfection. Additionally, the duration of transgene expression was extended for surface-mediated delivery relative to bolus delivery.


Within- and between-examiner agreement for two thyroid palpation techniques in healthy and hyperthyroid cats.
Thyroid gland palpation is an important aid for diagnosing feline hyperthyroidism in an early stage to prevent development of deleterious complications. Our objectives were to assess within- and between-examiner agreement for two thyroid gland palpation techniques in cats and to correlate palpation results with ultrasonographic thyroid measurements. Nine client-owned hyperthyroid (12.6 +/- 2.4 years) and 10 healthy control cats (7.4 +/- 5.4 years) entered this prospective study. Both thyroid glands of all cats were palpated twice by three blindfolded clinicians with the classic palpation technique [technique 1 (T1)] and the technique described by Norsworthy GD, Adams VJ, McElhaney MR, Milios JA [(2002a) Relationship between semi-quantitative thyroid palpation and total thyroxine concentration in cats with and without hyperthyroidism. Journal of Feline Medicine and Surgery 4, 139-143] [technique 2 (T2)]. A semi-quantitative score from 1 to 6 was assigned to the gland size. After clipping of the ventral cervical region, another palpation session followed by ultrasonography of the thyroid glands was performed. Average weighted kappa-values within- and between-examiners were 0.864 and 0.644 for T1 and 0.732 and 0.532 for T2. T1 did lead to significantly smaller within- (P=0.007) and between-examiner (P=0.048) differences than T2. Significant correlation coefficients (P<0.001) between the palpation scores of both techniques and ultrasonographic thyroid lobe length (T1: 0.43; T2: 0.38) were observed. No significant difference before and after clipping was found (T1: P=0.503; T2: P=0.607). The first time that all cats were palpated by either technique, significant score differences between control and hyperthyroid cats were observed both for T1 (P=0.002) and T2 (P=0.003). Both feline thyroid gland palpation techniques have good within- and between-examiner agreements. Based on this study, the classic palpation technique is preferred.


Design and baseline characteristics of the PerfectFit study: a multicenter cluster-randomized trial of a lifestyle intervention in employees with increased cardiovascular risk.
The prevalence of unhealthy lifestyles and preventable chronic diseases is high. They lead to disabilities and sickness absence, which might be reduced if health promotion measures were applied. Therefore, we developed the PerfectFit health promotion intervention with a "blended care"-approach, which consists of a web-based health risk assessment (HRA) including tailored and personalized advice, followed by motivational interviewing (MI). We hypothesize that adding MI to a web-based HRA leads to better health outcomes. The objective is to describe the design and baseline characteristics of the PerfectFit study, which is being conducted among employees with high cardiovascular risk in the military workforce, the police organization and an academic hospital. PerfectFit is a cluster randomized controlled trial, consisting of two arms. Based on cardiovascular risk profiling, done between 2012 and 2014, we included employees based on one or more risk factors and motivation to participate. One arm is the 'limited' health program (control) that consists of: (a) an HRA as a decision aid for lifestyle changes, including tailored and personalized advice, and pros and cons of the options, and (b) a newsletter every 3 months. The other arm is the 'extensive' program (intervention), which is additionally offered MI-sessions by trained occupational physicians, 4 face-to-face and 3 by telephone, and is offered more choices of health promotion activities in the HRA. During the follow-up period, participants choose the health promotion activities they personally prefer. After six and twelve months, outcomes will be assessed by online questionnaires. After twelve months the cardiovascular risk profiling will be repeated. The primary outcome is self-reported general health. Secondary outcomes are self-reported work ability, CVD-risk score, sickness absence, productivity loss at work, participation in health promotion activities, changes in lifestyle (smoking, alcohol consumption, physical activity, stress management) and body mass index. Furthermore, a process evaluation and an economic analysis will be performed. Additional coaching using MI is expected to be a key factor for success of the web-based HRA in employees with increased cardiovascular risk. This "blended care"-approach may be an essential strategy for effective health promotion activities. Dutch Trial Register by registration number NTR4894 , 14/11/2014.


Mutational status of the TP53 gene as a predictor of response and survival in patients with chronic lymphocytic leukemia: results from the LRF CLL4 trial.
TP53 mutations have been described in chronic lymphocytic leukemia (CLL) and have been associated with poor prognosis in retrospective studies. We aimed to address the frequency and prognostic value of TP53 abnormalities in patients with CLL in the context of a prospective randomized trial. We analyzed 529 CLL samples from the LRF CLL4 (Leukaemia Research Foundation Chronic Lymphocytic Leukemia 4) trial (chlorambucil v fludarabine with or without cyclophosphamide) at the time of random assignment for mutations in the TP53 gene. TP53 mutation status was correlated with response and survival data. Mutations of TP53 were found in 40 patients (7.6%), including 25 (76%) of 33 with 17p deletion and 13 (3%) of 487 without that deletion. There was no significant correlation between TP53 mutations and age, stage, IGHV gene mutations, CD38 and ZAP-70 expression, or any other chromosomal abnormality other than 17p deletion, in which concordance was high (96%). TP53 mutations were significantly associated with poorer overall response rates (27% v 83%; P < .001) and shorter progression-free survival (PFS) and overall survival (OS; 5-year PFS: 5% v 17%; 5-year OS: 20% v 59%; P < .001 for both). Multivariate analysis that included baseline clinical variables, treatment, and known adverse genetic factors confirmed that TP53 mutations have added prognostic value. TP53 mutations are associated with impaired response and shorter survival in patients with CLL. Analysis of TP53 mutations should be performed in patients with CLL who have progressive disease before starting first-line treatment, and those with mutations should be selected for novel experimental therapies.


Assisted suicide: clinical realities and ethical challenges.
The increasing attention to assisted suicide, as evidenced by recent legislation, initiatives, court decisions, and research, propels the issue to a new level of importance and urgency within society and the health professions. Nurses cannot help but be confronted by and struggle with the complex moral and professional quandaries related to assisted suicide. Critical care nurses must continue to evaluate the implications of the possible legalization of assisted suicide and to define the boundaries of morally acceptable professional practice. The challenges to the roles and responsibilities of critical care nurses that might occur if assisted suicide were legalized must be thoughtfully and responsibly explored.


Direct Observation of Photoinduced Tautomerization in Single Molecules at a Metal Surface.
Molecular switches are of fundamental importance in nature, and light is an important stimulus to selectively drive the switching process. However, the local dynamics of a conformational change in these molecules remain far from being completely understood at the single-molecule level. Here, we report the direct observation of photoinduced tautomerization in single porphycene molecules on a Cu(111) surface by using a combination of low-temperature scanning tunneling microscopy and laser excitation in the near-infrared to ultraviolet regime. It is found that the thermodynamically stable trans configuration of porphycene can be converted to the metastable cis configuration in a unidirectional fashion by photoirradiation. The wavelength dependence of the tautomerization cross section exhibits a steep increase around 2 eV and demonstrates that excitation of the Cu d-band electrons and the resulting hot carriers play a dominant role in the photochemical process. Additionally, a pronounced isotope effect in the cross section (∼100) is observed when the transferred hydrogen atoms are substituted with deuterium, indicating a significant contribution of zero-point energy in the reaction. Combined with the study of inelastic tunneling electron-induced tautomerization with the STM, we propose that tautomerization occurs via excitation of molecular vibrations after photoexcitation. Interestingly, the observed cross section of ∼10(-19) cm(2) in the visible-ultraviolet region is much higher than that of previously studied molecular switches on a metal surface, for example, azobenzene derivatives (10(-23)-10(-22) cm(2)). Furthermore, we examined a local environmental impact on the photoinduced tautomerization by varying molecular density on the surface and find substantial changes in the cross section and quenching of the process due to the intermolecular interaction at high density.


Choosing HIV Counseling and Testing Strategies for Outreach Settings: A Randomized Trial.
In surveys, clients have expressed preferences for alternatives to traditional HIV counseling and testing. Few data exist to document how offering such alternatives affects acceptance of HIV testing and receipt of test results. This randomized controlled trial compared types of HIV tests and counseling at a needle exchange and 2 bathhouses to determine which types most effectively ensured that clients received test results. Four alternatives were offered on randomly determined days: (1) traditional test with standard counseling, (2) rapid test with standard counseling, (3) oral fluid test with standard counseling, and (4) traditional test with choice of written pretest materials or standard counseling. Of 17,010 clients offered testing, 7014 (41%) were eligible; of those eligible, 761 (11%) were tested: 324 at the needle exchange and 437 at the bathhouses. At the needle exchange, more clients accepted testing (odds ratio [OR] = 2.3; P < 0.001) and received results (OR = 2.6; P < 0.001) on days when the oral fluid test was offered compared with the traditional test. At the bathhouses, more clients accepted oral fluid testing (OR = 1.6; P < 0.001), but more clients overall received results on days when the rapid test was offered (OR = 1.9; P = 0.01). Oral fluid testing and rapid blood testing at both outreach venues resulted in significantly more people receiving test results compared with traditional HIV testing. Making counseling optional increased testing at the needle exchange but not at the bathhouses.


Interference with p53 protein inhibits hematopoietic and muscle differentiation.
The involvement of p53 protein in cell differentiation has been recently suggested by some observations made with tumor cells and the correlation found between differentiation and increased levels of p53. However, the effect of p53 on differentiation is in apparent contrast with the normal development of p53-null mice. To test directly whether p53 has a function in cell differentiation, we interfered with the endogenous wt-p53 protein of nontransformed cells of two different murine histotypes: 32D myeloid progenitors, and C2C12 myoblasts. A drastic inhibition of terminal differentiation into granulocytes or myotubes, respectively, was observed upon expression of dominant-negative p53 proteins. This inhibition did not alter the cell cycle withdrawal typical of terminal differentiation, nor p21(WAF1/CIP1) upregulation, indicating that interference with endogenous p53 directly affects cell differentiation, independently of the p53 activity on the cell cycle. We also found that the endogenous wt-p53 protein of C2C12 cells becomes transcriptionally active during myogenesis, and this activity is inhibited by p53 dominant-negative expression. Moreover, we found that p53 DNA-binding and transcriptional activities are both required to induce differentiation in p53-negative K562 cells. Taken together, these data strongly indicate that p53 is a regulator of cell differentiation and it exerts this role, at least in part, through its transcriptional activity.


Integrating mobile-phone based assessment for psychosis into people's everyday lives and clinical care: a qualitative study.
Over the past decade policy makers have emphasised the importance of healthcare technology in the management of long-term conditions. Mobile-phone based assessment may be one method of facilitating clinically- and cost-effective intervention, and increasing the autonomy and independence of service users. Recently, text-message and smartphone interfaces have been developed for the real-time assessment of symptoms in individuals with schizophrenia. Little is currently understood about patients' perceptions of these systems, and how they might be implemented into their everyday routine and clinical care. 24 community based individuals with non-affective psychosis completed a randomised repeated-measure cross-over design study, where they filled in self-report questions about their symptoms via text-messages on their own phone, or via a purpose designed software application for Android smartphones, for six days. Qualitative interviews were conducted in order to explore participants' perceptions and experiences of the devices, and thematic analysis was used to analyse the data. Three themes emerged from the data: i) the appeal of usability and familiarity, ii) acceptability, validity and integration into domestic routines, and iii) perceived impact on clinical care. Although participants generally found the technology non-stigmatising and well integrated into their everyday activities, the repetitiveness of the questions was identified as a likely barrier to long-term adoption. Potential benefits to the quality of care received were seen in terms of assisting clinicians, faster and more efficient data exchange, and aiding patient-clinician communication. However, patients often failed to see the relevance of the systems to their personal situations, and emphasised the threat to the person centred element of their care. The feedback presented in this paper suggests that patients are conscious of the benefits that mobile-phone based assessment could bring to clinical care, and that the technology can be successfully integrated into everyday routine. However, it also suggests that it is important to demonstrate to patients the personal, as well as theoretical, benefits of the technology. In the future it will be important to establish whether clinical practitioners are able to use this technology as part of a personalised mental health regime.


Remote effects of kidney drug transporter OAT1 on gut microbiome composition and urate homeostasis.
The organic anion transporter OAT1 (SLC22A6, originally identified as NKT) is a multispecific transporter responsible for the elimination by the kidney of small organic anions that derive from the gut microbiome. Many are uremic toxins associated with chronic kidney disease (CKD). OAT1 is among a group of "drug" transporters that act as hubs in a large homeostatic network regulating interorgan and interorganismal communication via small molecules. The Remote Sensing and Signaling Theory predicts that genetic deletion of such a key hub in the network results in compensatory interorganismal communication (e.g., host-gut microbe dynamics). Recent metabolomics data from Oat1-KO mice indicate that some of the most highly affected metabolites derive from bacterial tyrosine, tryptophan, purine, and fatty acid metabolism. Functional metagenomic analysis of fecal 16S amplicon and whole-genome sequencing revealed that loss of OAT1 was impressively associated with microbial pathways regulating production of urate, gut-derived p-cresol, tryptophan derivatives, and fatty acids. Certain changes, such as alterations in gut microbiome urate metabolism, appear compensatory. Thus, Oat1 in the kidney appears to mediate remote interorganismal communication by regulating the gut microbiome composition and metabolic capability. Since OAT1 function in the proximal tubule is substantially affected in CKD, our results may shed light on the associated alterations in gut-microbiome dynamics.


Interface-mediated interactions: entropic forces of curved membranes.
Particles embedded in a fluctuating interface experience forces and torques mediated by the deformations and by the thermal fluctuations of the medium. Considering a system of two cylinders bound to a fluid membrane, we show that the entropic contribution enhances the curvature-mediated repulsion between the two cylinders. This is contrary to the usual attractive Casimir force in the absence of curvature-mediated interactions. For a large distance between the cylinders, we retrieve the renormalization of the surface tension of a flat membrane due to thermal fluctuations.


Fibronectin matrix-mediated cohesion suppresses invasion of prostate cancer cells.
Invasion is an important early step in the metastatic cascade and is the primary cause of death of prostate cancer patients. In order to invade, cells must detach from the primary tumor. Cell-cell and cell-ECM interactions are important regulators of cohesion--a property previously demonstrated to mediate cell detachment and invasion. The studies reported here propose a novel role for α5β1 integrin--the principle mediator of fibronectin matrix assembly (FNMA)--as an invasion suppressor of prostate cancer cells. Using a combination of biophysical and cell biological methods, and well-characterized prostate cancer cell lines of varying invasiveness, we explore the relationship between cohesion, invasiveness, and FNMA. We show that cohesion is inversely proportional to invasive capacity. We also show that more invasive cells express lower levels of α5β1 integrin and lack the capacity for FNMA. Cells were generated to over-express either wild-type α5 integrin or an integrin in which the cytoplasmic domain of α5 was replaced with that of α2. The α2 construct does not promote FNMA. We show that only wild-type α5 integrin promotes aggregate compaction, increases cohesion, and reduces invasion of the more aggressive cells, and that these effects can be blocked by the 70-kDa fibronectin fragment. We propose that restoring capacity for FNMA in deficient cells can increase tumor intercellular cohesion to a point that significantly reduces cell detachment and subsequent invasion. In prostate cancer, this could be of therapeutic benefit by blocking an early key step in the metastatic cascade.


Systemic vasculitis in patients with hepatitis C.
To analyze the main characteristics of patients infected with hepatitis C virus (HCV) presenting with different types of vasculitis syndrome. We retrospectively compared 2 groups of patients with HCV presenting with systemic vasculitis: 10 with biopsy proven polyarteritis nodosa-type systemic vasculitis (PAN, Group 1) and 7 with mixed cryoglobulinemia syndrome (MC, Group 2). Patients of Group 1 presented with different features than Group 2: life threatening systemic vasculitis (10 vs 0; p < 0.01), severe multifocal sensorimotor mononeuropathies versus distal moderate sensory polyneuropathies, malignant hypertension (5 vs 0; p = 0.04), cerebral angiitis (2 vs 0), ischemic abdominal pain (2 vs 0), kidney and liver microaneurisms (2 vs 0), increased erythrocyte sedimentation rate and C-reactive protein (7 vs 0; p < 0.01), renal insufficiency (5 vs 0; p = 0.04), HCV genotype 1b (3 vs 6; p = 0.06), and lower activity of chronic hepatitis (p = 0.02). Neuromuscular biopsies showed lesions of vasculitis in all patients, but the type of vasculitis was different in Group 1 compared to Group 2: medium size artery involvement (7 vs 0; p < 0.01), necrotizing vasculitis (10 vs 0; p < 0.01), and mononuclear cell infiltrate in perivascular areas (0 vs 7; p < 0.01). Using prednisone, plasma exchanges, and interferon-alpha, complete recovery was obtained in all PAN-type patients except one. In Group 2 patients, interferon-alpha did not have any effect on the peripheral neuropathy. HCV infection may be associated with different types of systemic vasculitis, i.e., polyarteritis nodosa or mixed cryoglobulinemia. Because of differences in clinical and pathological features and therapeutic strategy, PAN-type vasculitis should be distinguished from MC-type vasculitis in HCV patients.


Mucinous Breast Cancer: a Review Study of 5 Year Experience from a Hospital-Based Series of Cases.
Mucinous carcinoma (also known as colloid carcinoma) is a particular type of breast cancer characterized by the presence of extracellular mucin and is linked with a more favorable prognosis than invasive breast carcinoma of no special type. Mucinous carcinoma of the breast is an uncommon form of breast tumor, often presenting as a lobulated, moderately well circumscribed mass on mammography, sonography, and MRI imaging. It accounts for 1 to 7% of all breast cancers. Pure mucinous breast carcinomas are rare and account for about 2% of all primary breast carcinomas. Metastatic disease happens at a lower rate than in other types of invasive carcinoma. We present our 5 year experience with this particular pathology in a retrospective review study. We identified 25 patients with mixed and pure mucinous breast cancer, the tumor size varied greatly from 2 to 19 cm in diameter. A subset of mixed mucinous carcinomas (8 cases) showed neuroendocrine differentiation or other associated premalignant lessions. Mucinous carcinoma of the breast is a rare entity with a favorable prognosis due to low incidence of lymph node metastases. Pure mucinous breast carcinoma has an even rare.


Zebrafish (Danio rerio) embryo-larvae locomotor activity data analysis: Evaluating anxiolytic effects of the antidepressant compound citalopram.
Newly fertilized zebrafish (Danio rerio) embryos were exposed to increasing concentrations of the selective serotonin reuptake inhibitor (SSRI) citalopram from fertilization until six days post-fertilization (dpf). Locomotor activity data were acquired at six dpf using an automated ZebraBox® infrared tracking system. Individual (n = 32) locomotor activity was recorded during 75 minutes in total during alternating illumination conditions (0% light, i.e. dark periods, and 100% light, i.e. light periods). The first 15 minutes of the test consisted of a dark period, i.e an acclimatization phase. Afterward, six alternating light and dark periods were conducted. Individual zebrafish embryo-larvae locomotion was tracked and aggregated in ten-second bins. The dataset, containing nine locomotor-related quantified endpoints (factors), was parsed, analyzed, and visualized using R software. The dataset and its associated custom R script may be used to further explore locomotor activity outcomes (e.g. anxiolytic or anxiogenic properties) following exposure to citalopram or other neuroactive chemicals.


Prevention and reversal of premature endothelial cell senescence and vasculopathy in obesity-induced diabetes by ebselen.
Although the accelerated atherosclerosis and premature aging of the cardiovascular system in patients with metabolic syndrome have been appreciated, the mechanisms of their development and potential therapeutic interventions remain unresolved. Our previous studies implicated advanced glycosylation end products in development of premature senescence preventable with a peroxynitrite scavenger, ebselen. Therefore, the effect of ebselen on endothelial senescence and vasculopathy in a model of metabolic syndrome--Zucker diabetic rats (ZDF)--was investigated. Ebselen decreased the abundance of 3-nitrotyrosine-modified proteins in ZDF rats. A 6-fold increase in the number of senescent endothelial cells in 22-week-old ZDF was prevented by ebselen. Development of vasculopathy, as collectively judged by the acetylcholine-induced vasorelaxation, NO production, angiogenic competence, and number of circulating microparticles, was almost completely prevented when ebselen was administered from 8 to 22 weeks and partially reversed when the treatment interval was 13 to 22 weeks. In conclusion, premature senescence of endothelial cells is progressively rampant in ZDF rats and is associated with the signs of severe vasculopathy. In addition, prevention of premature senescence of vascular endothelium through controlled decrease in nitrotyrosine formation was chronologically associated with the amelioration of vasculopathy, lending support to the idea of the pathogenetic role of premature senescence of endothelial cells in diabetic macrovasculopathy.


Chronic oxidative stress compromises telomere integrity and accelerates the onset of senescence in human endothelial cells.
Replicative senescence and oxidative stress have been implicated in ageing, endothelial dysfunction and atherosclerosis. Replicative senescence is determined primarily by telomere integrity. In endothelial cells the glutathione redox-cycle plays a predominant role in the detoxification of peroxides. The aim of this study was to elucidate the role of the glutathione-dependent antioxidant system on the replicative capacity and telomere dynamics of cultured endothelial cells. Human umbilical vein endothelial cells were serially passaged while exposed to regular treatment with 0.1 microM tert-butyl hydroperoxide, a substrate of glutathione peroxidase, or 10 microM L-buthionine-[S,R]-sulphoximine, an inhibitor of glutathione synthesis. Both treatments induced intracellular oxidative stress but had no cytotoxic or cytostatic effects. Nonetheless, treated cultures entered senescence prematurely (30 versus 46 population doublings), as determined by senescence-associated beta-galactosidase staining and a sharp decrease in cell density at confluence. In cultures subjected to oxidative stress terminal restriction fragment (TRF) analysis demonstrated faster telomere shortening (110 versus 55 bp/population doubling) and the appearance of distinct, long TRFs after more than 15-20 population doublings. Fluorescence in situ hybridisation analysis of metaphase spreads confirmed the presence of increased telomere length heterogeneity, and ruled out telomeric end-to-end fusions as the source of the long TRFs. The latter was also confirmed by Bal31 digestion of genomic DNA. Similarly, upregulation of telomerase could not account for the appearance of long TRFs, as oxidative stress induced a rapid and sustained decrease in this activity. These findings demonstrate a key role for glutathione-dependent redox homeostasis in the preservation of telomere function in endothelial cells and suggest that loss of telomere integrity is a major trigger for the onset of premature senescence under mild chronic oxidative stress.


Multivariate optical computing using a digital micromirror device for fluorescence and Raman spectroscopy.
A multivariate optical computer has been constructed consisting of a spectrograph, digital micromirror device, and photomultiplier tube that is capable of determining absolute concentrations of individual components of a multivariate spectral model. We present experimental results on ternary mixtures, showing accurate quantification of chemical concentrations based on integrated intensities of fluorescence and Raman spectra measured with a single point detector. We additionally show in simulation that point measurements based on principal component spectra retain the ability to classify cancerous from noncancerous T cells.


To comply or not comply? A latent profile analysis of behaviours and attitudes during the COVID-19 pandemic.
How and why do people comply with protective behaviours during COVID-19? The emerging literature employs a variable-centered approach, typically using a narrow selection of constructs within a study. This study is the first to adopt a person-centred approach to identify complex patterns of compliance, and holistically examine underlying psychological differences, integrating multiple psychology paradigms and epidemiology. 1575 participants from Australia, US, UK, and Canada indicated their behaviours, attitudes, personality, cognitive/decision-making ability, resilience, adaptability, coping, political and cultural factors, and information consumption during the pandemic's first wave. Using Latent Profile Analysis, two broad groups were identified. The compliant group (90%) reported greater worries, and perceived protective measures as effective, whilst the non-compliant group (about 10%) perceived them as problematic. The non-compliant group were lower on agreeableness and cultural tightness-looseness, but more extraverted, and reactant. They utilised more maladaptive coping strategies, checked/trusted the news less, and used official sources less. Females showed greater compliance than males. By promoting greater appreciation of the complexity of behaviour during COVID-19, this research provides a critical platform to inform future studies, public health policy, and targeted behaviour change interventions during pandemics. The results also challenge age-related stereotypes and assumptions.


MMP13, Birc2 (cIAP1), and Birc3 (cIAP2), amplified on chromosome 9, collaborate with p53 deficiency in mouse osteosarcoma progression.
Osteosarcoma is the primary malignant cancer of bone and particularly affects adolescents and young adults, causing debilitation and sometimes death. As a model for human osteosarcoma, we have been studying p53(+/-) mice, which develop osteosarcoma at high frequency. To discover genes that cooperate with p53 deficiency in osteosarcoma formation, we have integrated array comparative genomic hybridization, microarray expression analyses in mouse and human osteosarcomas, and functional assays. In this study, we found seven frequent regions of copy number gain and loss in the mouse p53(+/-) osteosarcomas but have focused on a recurrent amplification event on mouse chromosome 9A1. This amplicon is syntenic with a similar chromosome 11q22 amplicon identified in several human tumor types. Three genes on this amplicon, the matrix metalloproteinase gene MMP13 and the antiapoptotic genes Birc2 (cIAP1) and Birc3 (cIAP2), show elevated expression in mouse and human osteosarcomas. We developed a functional assay using clonal osteosarcoma cell lines transduced with lentiviral short hairpin RNA vectors to show that down-regulation of MMP13, Birc2, or Birc3 resulted in reduced tumor growth when transplanted into immunodeficient recipient mice. These experiments revealed that high MMP13 expression enhances osteosarcoma cell survival and that Birc2 and Birc3 also enhance cell survival but only in osteosarcoma cells with the chromosome 9A1 amplicon. We conclude that the antiapoptotic genes Birc2 and Birc3 are potential oncogenic drivers in the chromosome 9A1 amplicon.


Neural Cell Adhesion Protein CNTN1 Promotes the Metastatic Progression of Prostate Cancer.
Prostate cancer metastasis is the main cause of disease-related mortality. Elucidating the mechanisms underlying prostate cancer metastasis is critical for effective therapeutic intervention. In this study, we performed gene-expression profiling of prostate cancer stem-like cells (PCSC) derived from DU145 human prostate cancer cells to identify factors involved in metastatic progression. Our studies revealed contactin 1 (CNTN1), a neural cell adhesion protein, to be a prostate cancer-promoting factor. CNTN1 knockdown reduced PCSC-mediated tumor initiation, whereas CNTN1 overexpression enhanced prostate cancer cell invasion in vitro and promoted xenograft tumor formation and lung metastasis in vivo. In addition, CNTN1 overexpression in DU145 cells and corresponding xenograft tumors resulted in elevated AKT activation and reduced E-cadherin (CDH1) expression. CNTN1 expression was not readily detected in normal prostate glands, but was clearly evident on prostate cancer cells in primary tumors and lymph node and bone metastases. Tumors from 637 patients expressing CNTN1 were associated with prostate cancer progression and worse biochemical recurrence-free survival following radical prostatectomy (P < 0.05). Collectively, our findings demonstrate that CNTN1 promotes prostate cancer progression and metastasis, prompting further investigation into the mechanisms that enable neural proteins to become aberrantly expressed in non-neural malignancies.


[Experimental validation of a semi-quantitative single-layer procedure for proton-spin tomographic imaging of regional cerebral perfusion].
The aim of the present study was to validate a simple MRI-procedure for semiquantitative assessment of regional cerebral blood flow. Unilateral cerebral ischaemia (30 minutes) in the territory of the middle cerebral artery was induced in 14 anesthetised rates. The MRI-experiment consisted in an intravenous bolus injection of gadolinium-DTPA, recording of the cerebral contrast kinetics with a T2*-weighted pulse sequence, and measurement of the maximal concentration change at a chosen reference point of time. To measure perfusion quantitatively, a microsphere technique, an accepted reference technique was used. With both methods a perfusion index related to the contralateral side was calculated. In all cases decreased perfusion was detected by the MRI technique. The perfusion indices correlated with a coefficient of correlation of r = 0.89 (p < 0.001). The results demonstrate that contrast-enhanced MRI with bolus injection can be implemented with clinical potential as a semiquantitative instrument for the assessment of cerebral perfusion. Regional cerebral blood volume and collateral blood flow may interfere with the estimate of blood flow.


Registered nurses with disabilities: legal rights and responsibilities.
The purpose of this legal case review and analysis was to determine what kinds of cases involving nurses with disabilities are typically brought to attorneys, which cases tend to be successful, and how and when a nurse with a disability should pursue legal action. The review used the standard legal case analysis method to analyze legal cases that have been brought by registered nurses (RNs) with physical or sensory disabilities from 1995 to 2013. The cases span the period following the enactment of the Americans With Disabilities Act (ADA) of 1990 through the ADA Amendments Act (ADAAA) of 2008. A nurse attorney reviewed the background material to find every case involving an RN with a disability, excluding those with mental health disabilities or substance abuse issues. Case analysis was conducted using standard legal case analysis procedures. Fifty-six cases were analyzed. The cases were categorized into five types of legal claims: (a) disability discrimination (84%); (b) failure to accommodate (46%); (c) retaliation (12.5%); (d) association (3.6%); and (e) hostile work environment (7%). The cases were largely unsuccessful, particularly those brought under the ADA instead of the ADAAA. The case analysis revealed that several cases brought by RNs with disabilities using the ADA might have been successful under the ADAAA. In addition, the case analysis has provided vital information for administrators, leaders, and clinical nurses regarding when a case is appropriate for legal action. These findings from this review will help nurses recognize when they are being treated in a discriminatory way in the workplace, what their legal rights and responsibilities are, and at what point they should pursue legal action. This review has relevance to all RNs working in clinical and academic settings who may have a congenital or acquired physical or sensory disability.


Molecular analysis of phosphate limitation in Geobacteraceae during the bioremediation of a uranium-contaminated aquifer.
Nutrient limitation is an environmental stress that may reduce the effectiveness of bioremediation strategies, especially when the contaminants are organic compounds or when organic compounds are added to promote microbial activities such as metal reduction. Genes indicative of phosphate-limitation were identified by microarray analysis of chemostat cultures of Geobacter sulfureducens. This analysis revealed that genes in the pst-pho operon, which is associated with a high-affinity phosphate uptake system in other microorganisms, had significantly higher transcript abundance under phosphate-limiting conditions, with the genes pstB and phoU upregulated the most. Quantitative PCR analysis of pstB and phoU transcript levels in G. sulfurreducens grown in chemostats demonstrated that the expression of these genes increased when phosphate was removed from the culture medium. Transcripts of pstB and phoU within the subsurface Geobacter species predominating during an in situ uranium-bioremediation field experiment were more abundant than in chemostat cultures of G. sulfurreducens that were not limited for phosphate. Addition of phosphate to incubations of subsurface sediments did not stimulate dissimilatory metal reduction. The added phosphate was rapidly adsorbed onto the sediments. The results demonstrate that Geobacter species can effectively reduce U(VI) even when experiencing suboptimal phosphate concentrations and that increasing phosphate availability with phosphate additions is difficult to achieve because of the high reactivity of this compound. This transcript-based approach developed for diagnosing phosphate limitation should be applicable to assessing the potential need for additional phosphate in other bioremediation processes.