Photophysical studies on lanthanide(III) chelates conjugated to Pittsburgh compound B as luminescent probes targeted to Aβ amyloid aggregates.
The photophysical properties of Eu3+ and Tb3+ complexes of DOTAGA and DO3A-monoamide conjugates of the Pittsburgh compound B (PiB) chromophore, prepared using linkers of different lengths and flexibilities, and which form stable negatively charged (LnL1), and uncharged (LnL2) complexes, respectively, were studied as potential probes for optical detection of amyloid aggregates. The phenylbenzothiazole (PiB) moiety absorbs light at wavelengths longer than 330 nm with a high molar absorption coefficient in both probes, and acts as an antenna in these systems. The presence of the luminescent Ln3+ ion quenches the excited states of PiB through an energy transfer process from the triplet state of PiB to the metal centre, and structured emission is seen from Eu3+ and Tb3+. The luminescence study indicates the presence of a 5D4 → T1 back transfer process in the Tb3+ complexes. It also provides insights on structural properties of the Eu3+ complexes, such as the high symmetry environment of the Eu3+ ion in a single macrocyclic conformation and the presence of one water molecule in its inner coordination sphere. The overall quantum yield of luminescence of EuL1 is higher than for EuL2. However, their low values reflect the low overall sensitization efficiency of the energy transfer process, which is a consequence of the large distances between the metal center and the antenna, especially in the EuL2 complex. DFT calculations confirmed that the most stable conformation of the Eu3+ complexes involves a combination of a square antiprismatic (SAP) geometry of the chelate and an extended conformation of the linker. The large calculated average distances between the metal center and the antenna point to the predominance of the Förster energy transfer mechanism, especially for EuL2. This study provides insights into the behavior of amyloid-targeted Ln3+ complexes as optical probes, and contributes towards their rational design.


What are the optimal cut-off points of anthropometric indices for prediction of overweight and obesity? Predictive validity of waist circumference, waist-to-hip and waist-to-height ratios.
Background: Planning for obesity prevention is an important global health priority. Our aim in this study was to find the optimal cut-off points of waist circumference (WC), waist- to- hipratio (WHR) and waist- to- height ratio (WHtR), as three anthropometric indices, for prediction of overweight and obesity. We also aimed to compare the predictive ability of these indices to introduce the best choice. Methods: In this cross-sectional study, a total of 500 subjects were investigated. Anthropometric indicators were measured using a standard protocol. We considered body mass index (BMI) as the simple and most commonly used index for measuring general obesity as the comparison indicator in the present study to assess the diagnostic value for other reported obesity indices.We also performed receiver operating characteristic (ROC) curve analysis to define the optimal cut-off points of the anthropometric indicators and the best indices for overweight and obesity. Results: The proposed optimal cut-offs for WC, WHtR, and WHR were 84 cm, 0.48 and 0.78 for women and 98 cm, 0.56 and 0.87 for men, respectively. The area under the ROC curve ofWHtR (women: AUC=0.97, 95% CI: 0.96-0.99 vs. men: AUC=0.97, 95%CI: 0.96-0.99) and WC(women: AUC=0.97, 95% CI, 0.95-0.99 vs. men: AUC=0.98, 95% CI: 0.97-0.99) were greater than WHR (women: AUC=0.79, 95% CI =0.74-0.85 vs. men: AUC=0.84, 95% CI=0.79-0.88). Conclusion: This study demonstrated that the WC and WHtR indicators are stronger indicators compared to the others. However, further studies using desirable and also local cutoffs against more accurate techniques for body fat measurement such as computerized tumor (CT) scans and dual-energy x-ray absorptiometry (DEXA) are required.


The caspase-related protease separase (extra spindle poles) regulates cell polarity and cytokinesis in Arabidopsis.
Vesicle trafficking plays an important role in cell division, establishment of cell polarity, and translation of environmental cues to developmental responses. However, the molecular mechanisms regulating vesicle trafficking remain poorly understood. Here, we report that the evolutionarily conserved caspase-related protease separase (extra spindle poles [ESP]) is required for the establishment of cell polarity and cytokinesis in Arabidopsis thaliana. At the cellular level, separase colocalizes with microtubules and RabA2a (for RAS genes from rat brainA2a) GTPase-positive structures. Separase facilitates polar targeting of the auxin efflux carrier PIN-formed2 (PIN2) to the rootward side of the root cortex cells. Plants with the radially swollen4 (rsw4) allele with compromised separase activity, in addition to mitotic failure, display isotropic cell growth, perturbation of auxin gradient formation, slower gravitropic response in roots, and cytokinetic failure. Measurements of the dynamics of vesicle markers on the cell plate revealed an overall reduction of the delivery rates of KNOLLE and RabA2a GTPase in separase-deficient roots. Furthermore, dissociation of the clathrin light chain, a protein that plays major role in the formation of coated vesicles, was slower in rsw4 than in the control. Our results demonstrate that separase is a key regulator of vesicle trafficking, which is indispensable for cytokinesis and the establishment of cell polarity.


Daily home-based spirometry during withdrawal of inhaled corticosteroid in severe to very severe chronic obstructive pulmonary disease.
The WISDOM study (NCT00975195) reported a change in lung function following withdrawal of fluticasone propionate in patients with severe to very severe COPD treated with tiotropium and salmeterol. However, little is known about the validity of home-based spirometry measurements of lung function in COPD. Therefore, as part of this study, following suitable training, patients recorded daily home-based spirometry measurements in addition to undergoing periodic in-clinic spirometric testing throughout the study duration. We subsequently determined the validity of home-based spirometry for detecting changes in lung function by comparing in-clinic and home-based forced expiratory volume in 1 second in patients who underwent stepwise fluticasone propionate withdrawal over 12 weeks versus patients remaining on fluticasone propionate for 52 weeks. Bland-Altman analysis of these data confirmed good agreement between in-clinic and home-based measurements, both across all visits and at the individual visits at study weeks 6, 12, 18, and 52. There was a measurable difference between the forced expiratory volume in 1 second values recorded at home and in the clinic (mean difference of -0.05 L), which may be due to suboptimal patient effort in performing unsupervised recordings. However, this difference remained consistent over time. Overall, these data demonstrate that home-based and in-clinic spirometric measurements were equally valid and reliable for assessing lung function in patients with COPD, and suggest that home-based spirometry may be a useful tool to facilitate analysis of changes in lung function on a day-to-day basis.


Enterocyte-specific inactivation of SIRT1 reduces tumor load in the APC(+/min) mouse model.
SIRT1 is a mammalian NAD(+)-dependent histone deacetylase implicated in metabolism, development, aging and tumorigenesis. Prior studies that examined the effect of enterocyte-specific overexpression and global deletion of SIRT1 on polyp formation in the intestines of APC(+/min) mice, a commonly used model for intestinal tumorigenesis, yielded conflicting results, supporting either tumor-suppressive or tumor-promoting roles for SIRT1, respectively. In order to resolve the controversy emerging from these prior in vivo studies, in the present report we examined the effect of SIRT1 deficiency confined to the intestines, avoiding the systemic perturbations such as growth retardation seen with global SIRT1 deletion. We crossed APC(+/min) mice with mice bearing enterocyte-specific inactivation of SIRT1 and examined polyp development in the progeny. We found that SIRT1-inactivation reduced total polyp surface (9.3 mm(2) vs. 23.3 mm(2), p = 0.01), average polyp size (0.24 mm(2) vs. 0.51 mm(2), p = 0.005) and the number of polyps >0.5 mm in diameter (14 vs. 23, p = 0.04), indicating that SIRT1 affects both the number and size of tumors. Additionally, tumors in SIRT1-deficient mice exhibited markedly increased numbers of cells undergoing apoptosis, suggesting that SIRT1 contributes to tumor growth by enabling survival of tumor cells. Our results indicate that SIRT1 acts as a tumor promoter in the APC(+/min) mouse model of intestinal tumorigenesis.


The Medical Emergency Team system: a two hospital comparison.
To compare activity and outcomes of a mature Medical Emergency Team (MET) in two hospitals. SETTING AND POPULATIONS: A Tertiary Referral Hospital (TRH) and a Metropolitan General Hospital (MGH) who combined have approximately 82,000 admissions annually with 38,000 patients meeting the eligibility criteria. The population included all admissions to the two hospitals aged 15 years and over with a stay>1 day (12 months period). Admissions that had a MET call originating in general wards were defined as Admissions Associated with a MET call (AAMET). A retrospective analysis of MET call audit forms, a Death Review database, and routinely collected hospital data for the period 1st October 2004 to 30th September 2005, inclusive. Chronic morbidity was calculated as a Charlson Index (CI) score over previous visits and admissions using ICD10 & ICD9 diagnosis and procedure codes. There were 633 and 349 AAMETs. The incidence rates (MET calls/1000 admissions) were 37.6 and 34.1. They were associated with being elderly; males; higher CI scores; surgical admissions, Emergency Department (ED) admissions, and longer length of stay (LOS). A systolic BP<90mm Hg, and "worried" were the most frequent MET call criteria. There were 27 (4.3%) and 9 (2.6%) deaths following a MET call, of these 17 and 5 had Cardiac Arrest (CA) as the reason for the call. Death occurred for 192 and 54 AAMETs, only 38 (20%) and 14 (26%) were Do Not Attempt Resuscitation (DNAR) deaths. One hundred and forty-seven (23.2%) and eighty-seven (24.9%) AAMETs had a MET call within 24h of transfer from a critical care area; the proportions of transfers differed significantly between the two hospitals. A well established MET system identified similar AAMET populations from two different hospital populations. Sick, elderly, and surgical rather than medical patients were associated with MET activity in both hospitals. Further research is needed to estimate the impact of increased monitoring and interventions on patient outcomes, and the role of MET teams in end of life decision-making.


Very high speed cw digital holographic interferometry.
It is reported for the first time the use of a very high speed camera in digital holographic interferometry with an out of plane sensitivity setup. The image plane holograms of a spherical latex balloon illuminated by a cw laser were acquired at a rate of 4000 frames per second, representing a time spacing between holograms of 250 microseconds, for 512 x 512 pixels at 8 bits resolution. Two types of tests were accomplished for a proof of principle of the technique, one with no constrains on the object which meant random movements due to non controlled environmental air currents, and the other with specific controlled conditions on the object. Results presented correspond to a random sample of sequential digital holograms, chosen from a 1 second exposure, individually Fourier processed in order to perform the usual comparison by subtraction between consecutive pairs thus obtaining the phase map of the object out of plane displacement, shown as a movie.


Nasopharyngeal vs. adenoid cultures in children undergoing adenoidectomy: prevalence of bacterial pathogens, their interactions and risk factors.
Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus colonization of the adenoids and nasopharynx in 103 preschool children who underwent adenoidectomy for recurrent upper respiratory tract infections was examined. Bacterial interactions and risk factors for bacterial colonization of the nasopharynx and adenoids, separately, were analysed statistically. The prevalence of simultaneous isolation from both anatomical sites was 45·6% for S. pneumoniae, 29·1% for H. influenzae, 15·5% for M. catarrhalis and 18·4% for S. aureus. Three pathogens were significantly more frequent together from adenoid samples; nasopharyngeal swabs more often yielded a single organism, but without statistical significance. M. catarrhalis and S. aureus significantly more frequently co-existed with S. pneumoniae and H. influenzae than with each other and a positive association of S. pneumoniae and H. influenzae in adenoid samples was evident. Several differences between risk factors for nasopharyngeal and adenoid colonization by the individual pathogens were observed. We conclude that the adenoids and nasopharynx appear to differ substantially in colonization by pathogenic microbes but occurrence of H. influenzae and S. pneumoniae in the nasopharynx could be predictive of upper respiratory tract infections.


The influence of psychological state and motivation on brain-computer interface performance in patients with amyotrophic lateral sclerosis - a longitudinal study.
The current study investigated the effects of psychological well-being measured as quality of life (QoL), depression, current mood and motivation on brain-computer interface (BCI) performance in amyotrophic lateral sclerosis (ALS). Six participants with most advanced ALS were trained either for a block of 20 sessions with a BCI based on sensorimotor rhythms (SMR) or a block of 10 sessions with a BCI based on event-related potentials, or both. Questionnaires assessed QoL and severity of depressive symptoms before each training block and mood and motivation before each training session. The SMR-BCI required more training than the P300-BCI. The information transfer rate was higher with the P300-BCI (3.25 bits/min) than with the SMR-BCI (1.16 bits/min). Mood and motivation were related to the number of BCI sessions. Motivational factors, specifically challenge and mastery confidence, were positively related to BCI performance (controlled for the number of sessions) in tow participants, while incompetence fear was negatively related with performance in one participant. BCI performance was not related to motivational factors in three other participants nor to mood in any of the six participants. We conclude that motivational factors may be related to BCI performance in individual subjects and suggest that motivational factors and well-being should be assessed in standard BCI protocols. We also recommend using P300-based BCI as first choice in severely paralyzed patients who present with a P300 evoked potential.


Hypoxia inducible factor-stabilizing bioactive glasses for directing mesenchymal stem cell behavior.
Oxygen tension is a known regulator of mesenchymal stem cell (MSC) plasticity, differentiation, proliferation, and recruitment to sites of injury. Materials capable of affecting the MSC oxygen-sensing pathway, independently of the environmental oxygen pressure, are therefore of immense interest to the tissue engineering (TE) and regenerative medicine community. In this study, we describe the evaluation of the effect of hypoxia inducible factor (HIF)-stabilizing bioactive glasses (BGs) on human MSCs. The dissolution products from these hypoxia-mimicking BGs stabilized HIF-1α in a concentration-dependent manner, altered cell proliferation and metabolism, and upregulated a number of genes involved in the hypoxic response (HIF1A, HIF2A, and VHL), MSC survival (SAG and BCL2), extracellular matrix remodeling (MMP1), and angiogenesis (VEGF and PDGF). These HIF-stabilizing materials can therefore be used to improve MSC survival and enhance regeneration in a number of TE strategies.


Selected nutritional habits of teenagers associated with overweight and obesity.
A balanced diet is at the heart of healthy growth and development of children and youth, whereas inappropriate eating habits considerably influence the incidence of disorders connected with nutrition, including overweight and obesity. This paper aims at studying nutritional factors of 18-year-old secondary school youth in the urban environment and their effect on the incidence of overweight and obesity. The survey was conducted among 1,999 secondary school students chosen at random. The research tool consisted an original survey questionnaire. The measurements of respondents' height and body mass provided data for calculating the body mass index. The percentage of youth with deficient body mass was estimated at 8.4%. The percentage of normal weight students in the surveyed group was estimated at 77.6%. Overweight and obesity characterized 14.0% of the total number. As many as 21.8% of overweight and obese respondents would eat one or two meals as opposed to 16.8% of normal weight students. Three-fourths of the surveyed students would eat breakfast regardless of their nutritional habits. Lunch is eaten by 52.9% of normal weight 18-year-olds and 46.1% of overweight and obese students. The analysis of mealtimes suggests that overweight and obese students would have their breakfast and dinner at later hours than the rest of the surveyed. More than half of the participating students failed to eat lunch (53.9%), and one in four students within this group resigned from supper. Girls would eat fruit and vegetables more frequently than boys several times a day. The percentage of persons in the surveyed groups who would eat fast foods on a daily basis was similar regardless of their nutritional status. Sweetened carbonated beverages would be drunk more often by overweight and obese boys (81.2%) as compared with boys with proper body mass (75.8%). The same type of beverages would be popular with two-thirds of girls, and this result was similar regardless of their nutritional status. About 44.2% of overweight and obese girls and 20% of girls with proper body mass attempted to lose weight, and 5.7% of boys tried to go on a diet. Eating limitations were declared by 16.5% of overweight and obese boys and ca. 3% of normal weight boys. Within the surveyed group of youth, it was possible to indicate eating errors primarily consisting in irregular eating, too low a number of meals during the day, particularly skipping breakfast, which took place more frequently among overweight and obese students rather than normal weight ones. The survey points to the insufficient intake of vegetables and fruit as opposed to salty and sweet meals. It is essential to convey the knowledge on the causes of overweight and obesity as well as rules of a healthy diet as factors preventing civilization diseases.


Intragenic and intergenic recombination between temperature-sensitive mutants of vaccinia virus.
The frequency of recombination for a complete set of two-factor crosses between vaccinia virus mutations separated by distances of between 54 and 10692 bp was determined. The results show that in intragenic crosses there is a linear relationship between the recombination frequency observed and distances between the mutations of up to 700 bp. However, no length dependence of the recombination frequency in intergenic crosses with a distance between mutations of 328 to greater than 10000 bp is observed. We attribute this lack of dependence to the high rate of viral DNA interchange, which makes some step other than the cross-over event rate-limiting. Furthermore, we believe that the observed difference in recombination frequency between inter- and intragenic recombination is due to complementation between temperature-sensitive mutants at the permissive temperature.


Oxidative stress and intraerythrocytic concentrations of sodium and potassium in diabetic dogs.
Oxidative stress parameters; thiobarbituric acid reaction substances (RBC-TBARS), catalase (RBC-CAT) and reduced glutathione (RBC-GSH) and the intraerythrocytic concentrations of electrolytes; sodium and potassium (RBC-Na and RBC-K) were determined in 18 well- controlled (WC) and 22 poorly-controlled diabetic mellitus (DM). Dogs with DM had significant higher blood glucose concentration (P < 0.001), haemoglobin A1c (P < 0.01) and fructosamine (P < 0.001) compared to normal healthy dogs (n = 19). Diabetic dogs in both groups had higher RBC-CAT (P < 0.05) while RBC-TBARS were higher significantly only in poorly-controlled DM group (P < 0.05). The RBC-K was significantly higher in both DM groups (P < 0.001). No changes in RBC-GSH and RBC-Na were found between DM and control healthy dogs. By linear regression analysis, the relationship were found between degree of diabetic mellitus and RBC-CAT, RBC-TBARS, RBC-Na and RBC-K. The relationship was also found between oxidative stress parameters and intraerythrocytic K+. The results suggest that in diabetic dogs, oxidative stress occurs which related to the severity of disease and may affect potassium homeostasis.


Organizational and Financial Analysis of Polish Tobacco Control Program in 2000-2018.
In accordance with the provisions of the WHO Framework Convention on Tobacco Control (FCTC), each country shall promote and strengthen public awareness of tobacco control issues (Article 12). Many parties to the FCTC have adopted national tobacco control programs to organize their tobacco control activities. The aim of our study was to analyze the organization and funding of the Polish Tobacco Control Program in years 2000-2018. Document analysis of The Program and reports from its implementation were performed in accordance to the Agency for Health Technology Assessment in Poland (AHTAPol) recommendations and the WHO FCTC guidelines for Article 12 implementation. Spending was also analyzed. The study showed both inadequate planning of and funding for Polish Tobacco Control Program. The Program was developed without use of best practices detailed in the WHO FCTC guidelines as well as in national guidelines prepared by AHTAPol. The experience of Poland shows that although earmarking tobacco taxes has existed in the law, it has been largely ineffective due to the poor Tobacco Control Program design and insufficient funding resulting from a poor execution of the earmarking law. This may be a warning to other countries to strive to create law, compliance with which can be verified and controlled.


Antibodies against M-Type Phospholipase A2 Receptor May Predict Treatment Response and Outcome in Membranous Nephropathy.
Anti-phospholipase A2 receptor (PLA2R) antibodies are specific to the diagnosis of primary membranous nephropathy (pMN). The prevalence of positive antibodies varies among different cohorts. Still there is discrepancy in regard to the association between antibody levels and clinical courses, and the prognostic value of antibodies to treatment responses and kidney outcomes. Three hundred fifty-nine consecutive kidney biopsy-proven pMN patients were enrolled. Anti-PLA2R antibodies were detected by immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA). The positive rate of anti-PLA2R antibodies in pMN was 65.2% (234/359) by IFA and 56.3% (202/359) by ELISA. The antibody level presented positive correlation with urinary protein excretion (r = 0.164, p = 0.002). Detectable antibodies and a higher level of proteinuria were independent risk factors to no-remission after treatments (OR 3.15, p = 0.004; OR 1.11, p = 0.006) and were independent risk factors to no-spontaneous remission (OR 2.20, p = 0.011; OR 1.36, p < 0.001). A higher level of antibodies (hazard ratio 1.002, p = 0.019) was the independent risk factor to kidney dysfunction during follow-up. The antibodies turned negative in 42 out of 52 (80.8%) patients who achieved clinical remission, while they remained positive in all patients of the no-response category (p < 0.001). We documented correlations between anti-PLA2R antibody levels and clinical severity in this large Chinese pMN cohort. Antibody positivity and higher antibody level might predict treatment responses and kidney outcomes of pMN.


Progressive degeneration of dopaminergic neurons through TRP channel-induced cell death.
Progressive neurodegenerative diseases are among the most frequently occurring aging-associated human pathologies. In a screen for Caenorhabditis elegans mutant animals that lack their normal complement of dopaminergic neurons, we identified two strains with progressive loss of dopaminergic neurons during postembryonic life. Through whole-genome sequencing we show that both strains harbor dominant (d), gain-of-function mutations in the Transient Receptor Potential (TRP) mechanosensory channel trp-4, a member of the invertebrate and vertebrate TRPN-type of the TRP family channels. Gain-of-function mutations in TRP channels have not been previously implicated in dopaminergic neuronal degeneration. We show that trp-4(d) induces cell death in dopamine neurons through a defined, calcium-related downstream pathway.


[Cervical adenopathies in children].
Discovering a neck lump is always source of parental anxiety and of medical concern. In most cases, some clinical data and certain anatomical aspects indicate the diagnostic options and the appropriate management.


Acute cholecystitis: when is the best time for laparoscopic cholecystectomy?
Laparoscopic cholecystectomy is no more an elective procedure. The question is when to perform laparoscopic cholecystectomy in the face of acute cholecystitis. The last decade (1995-2004) 297 patients had a laparoscopic cholecystectomy for acute cholecystitis. One hundred forty six of them were operated in the first 24-48h after the onset of symptoms (group I), 68 were operated in less than 4 weeks time after the attack of the acute cholecystitis (group II), while the rest 83 patients had a history of acute cholecystitis at least four weeks before their elective laparoscopic cholecystectomy (group III). Analysis of the operative time, complications and hospital stay showed that laparoscopic cholecystectomy in acute cholecystitis is the recommended surgical procedure. The success of the operation depends on the degree of the inflammatory changes in the gallbladder and the expertise of the operator both in emergency and laparoscopic surgery. Timing of the operation is crucial to executing a successful procedure. The operation is easier, faster and safer when performed in first 4-5 days of the onset of symptoms.


Controlled studies of a new microprocessor-based portable infusion pump.
A new microprocessor-controlled portable infusion pump, the Pancretec Provider IV 2000 TM was tested in vitro and in vivo in cancer patients. The Provider is a rotary peristaltic, battery-powered pump capable of flow rates of 0.2 to 83 ml/hour with delivery volumes up to 1999 ml. Two programming modes are available: intermittent infusion and continuous infusion. Bench tests showed the flow rate accuracy to be within 96% of the desired rate. Flow rate precision was similarly excellent at +/- 2%. Clinical studies were performed in 14 ambulatory patients receiving continuous infusion antineoplastic or analgesic drugs over 5-60 days as outpatients. The majority of infusions delivered fluoropyrimidines via indwelling central venous access ports. Two of the patients received long term (60 days) continuous infusion of analgesics for pain control. Flow rate accuracy with the pumps was within +/- 5% in 90% of the 27 infusion courses (244 patient-days of continuous infusion therapy). A significant therapy deviation (interruption 10% of the desired course) occurred in three instances. One related to a procedural error (incomplete cartridge insertion into the pump), two were caused by fluid leakage which interrupted pump function. Defect alarms (both visual and audible) operated in both instances. A KVO flow rate of 0.1 ml/hr was also found to be adequate to maintain catheter patency in peripheral veins over a 24-hour period in two normal volunteers. We conclude that the Provider pump is an accurate, reliable and state of the art infusion system with wide clinical applicability.


The Current State of Behavioral Health Quality Measures: Where Are the Gaps?
This review examined the extent to which existing behavioral health quality measures address the priority areas of the National Behavioral Health Quality Framework (NBHQF) as well as the extent to which the measures have received National Quality Forum endorsement and are used in major reporting programs. This review identified behavioral health quality measures in widely used measure inventories, including the National Quality Measures Clearinghouse, National Quality Forum, and the Center for Quality Assessment in Mental Health. Additional measures were identified through outreach to federal agencies. Measures were categorized by type, condition, target population, data source, reporting unit, endorsement status, and use in reporting programs. The review identified 510 measures. Nearly one-third of these measures address broad mental health or substance use conditions rather than a specific condition or diagnosis. Seventy-two percent are process measures. The most common data source for measures is administrative claims, and very few measures rely on electronic health records or surveys. Fifty-three (10%) measures have received National Quality Forum (NQF) endorsement, and 28 (5%) unique measures are used in major quality reporting programs. Several subdomains of the NBHQF, such as treatment intensification, financial barriers to care, and continuity of care, lack measures that are NQF endorsed. Despite the wide array of behavioral health quality measures, relatively few have received endorsement or are used in reporting programs. Future efforts should seek to fill gaps in measurement and to identify the most salient and strongest measures in each priority area.


Comparison of procalcitonin with C-reactive protein, interleukin 6 and interferon-alpha for differentiation of bacterial vs. viral infections.
Procalcitonin (PCT) concentration increases in bacterial infections but remains low in viral infections and inflammatory diseases. The change is rapid and the molecule is stable, making it a potentially useful marker for distinguishing between bacterial and viral infections. PCT concentration was determined with an immunoluminometric assay on plasma collected at admission in 360 infants and children hospitalized for bacterial or viral infection. It was compared with C-reactive protein (CRP), interleukin 6 and interferon-alpha measured on the same sample. The mean PCT concentration was 46 microg/l (median, 17.8) in 46 children with septicemia or bacterial meningitis. PCT concentration was > 1 microg/l in 44 of 46 in this group and in 59 of 78 children with a localized bacterial infection who had a negative blood culture (sensitivity, 83%). PCT concentration was > 1 microg/l in 16 of 236 children with a viral infection (specificity, 93%). PCT concentration was low in 9 of 10 patients with inflammatory disease and fever. A CRP value > or =20 mg/l was observed in 61 of 236 patients (26%) with viral infection and in 105 of 124 patients (86%) with bacterial infection. IL-6 was > 100 pg/ml in 14% of patients infected with virus and in 53% with bacteria. A secretion of interferon-alpha was found in serum in 77% of viral infected patients and in 8.6% of bacterial infected patients. In this study a PCT value of 1 microg/l or greater had better specificity, sensitivity and predictive value than CRP, interleukin 6 and interferon-alpha in children for distinguishing between viral and bacterial infections. PCT values are higher in invasive bacterial infections, but the cutoff value of 1 microg/l indicates the severity of the disease in localized bacterial infection and helps to decide antibiotic treatment in emergency room. PCT may be useful in an emergency room for differentiation of bacterial vs. viral infections in children and for making decisions about antibiotic treatments.


Elevated serum level of interleukin-32α in the patients with myasthenia gravis.
A new cytokine, interleukin-32 (IL-32), has been implicated in the pro-inflammatory immune responses in several autoimmune disorders, such as rheumatoid arthritis and inflammatory bowel diseases. Myasthenia gravis (MG) is a well-characterized autoimmune disease directed at the postsynaptic acetylcholine receptor (AChR) or end plate of the neuromuscular junction. IL-32 is a cytokine that induces tumor necrosis factor (TNF)-α, IL-6, IL-1β, and chemokine. IL-6, TNF-α, and IL-2 are related to the pathogenesis and immunoregulation of MG. The gene expression of IL-32 is increased in human natural killer (NK) cells and T lymphocytes when stimulated by IL-2 or mitogen. NK cells influence the development of experimental autoimmune MG (EAMG) and possibly MG. The aim of this study was to examine whether IL-32α levels are increased in patients with MG and to investigate the relationship between IL-32α levels and disease activity in human MG. Serum IL-32α levels were significantly higher in the MG patients (p = 0.03): 460.07 ± 192.30 pg/mL in MG patients and 248.45 ± 188.42 pg/mL in the healthy control group. Although there was no significant statistical difference, serum IL-32α levels of patients with both anti-AChR binding and blocking antibodies trended to be higher than those without either antibodies (521.56 ± 212.92 pg/mL vs. 339.52 ± 182.78 pg/mL, p = 0.16). IL-32α serum levels tended to decrease with clinical improvement in generalized MG. This study suggests the possibility that IL-32 might contribute to MG pathogenesis or immunoregulation.


Participatory testing and reporting in an environmental-justice community of Worcester, Massachusetts: a pilot project.
Despite indoor home environments being where people spend most time, involving residents in testing those environments has been very limited, especially in marginalized communities. We piloted participatory testing and reporting that combined relatively simple tests with actionable reporting to empower residents in Main South/Piedmont neighborhoods of Worcester, Massachusetts. We answered: 1) How do we design and implement the approach for neighborhood and household environments using participatory methods? 2) What do pilot tests reveal? 3) How does our experience inform testing practice? The approach was designed and implemented with community partners using community-based participatory research. Residents and researchers tested fourteen homes for: lead in dust indoors, soil outdoors, paint indoors and drinking water; radon in basement air; PM2.5 in indoor air; mold spores in indoor/outdoor air; and drinking water quality. Monitoring of neighborhood particulates by residents and researchers used real-time data to stimulate dialogue. Given the newness of our partnership and unforeseen conflicts, we achieved moderate-high success overall based on process and outcome criteria: methods, test results, reporting, lessons learned. The conflict burden we experienced may be attributable less to generic university-community differences in interests/culture, and more to territoriality and interpersonal issues. Lead-in-paint touch-swab results were poor proxies for lead-in-dust. Of eight units tested in summer, three had very high lead-in-dust (>1000 microg/ft2), six exceeded at least one USEPA standard for lead-in-dust and/or soil. Tap water tests showed no significant exposures. Monitoring of neighborhood particulates raised awareness of environmental health risks, especially asthma. Timely reporting back home-toxics' results to residents is ethical but it must be empowering. Future work should fund the active participation of a few motivated residents as representatives of the target population. Although difficult and demanding in time and effort, the approach can educate residents and inform exposure assessment. It should be considered as a core ingredient of comprehensive household toxics' testing, and has potential to improve participant retention and the overall positive impact of long-term environmental health research efforts.


[Sleep--wakefulness cycle of cold-blooded vertebrates according to the results of continuous contact-free recording of the heart rate and motor activity].
Using continuous non-contact registration of the heart rate and motor activity, separate states were revealed in awakefulness--sleep cycle of various poikilotherms (the catfish Ictalurus nebulosus, frog Rana temporaria, tortoise Emys orbicularis). The method employed allowed also to classify motor automatic activity which was observed in earlier experiments at the background of sleep-like forms of the resting state. Two basic types of these automaticities were distinguished. Comparison of the data obtained on poikilothermic animals with those obtained on homoiothermic ones (the ground squirrel Citellus parryi) indicates the validity of using the heart rate and motor activity for detection of the awakefulness-sleep cycle.


Performance properties of the population bioequivalence approach for in vitro delivered dose for orally inhaled respiratory products.
Regulatory agencies, industry, and academia have acknowledged that in vitro assessments serve a role in establishing bioequivalence for second-entry drug product approvals as well as innovator post-approval drug product changes. For orally inhaled respiratory products (OIPs), the issues of correctly analyzing in vitro data and interpreting the results within the broader context of therapeutic equivalence have garnered significant attention. One of the recommended statistical tests for in vitro data is the population bioequivalence method (PBE). The current literature for assessing the PBE statistical approach for in vitro data assumes a log normal distribution. This paper focuses on an assessment of that assumption for in vitro delivered dose. Concepts in development of a statistical model are presented. The PBE criterion and hypotheses are written for the case when data follows a normal distribution, rather than log normal. Results of a simulation study are reported, characterizing the performance of the PBE approach when data are expected to be normally distributed, rather than log normal. In these cases, decisions using the PBE approach are not consistent for the same absolute mean difference that the test product is from the reference product. A conclusion of inequivalency will occur more often if the test product dose is lower than the reference product for the same deviation from target. These features suggest that more research is needed for statistical equivalency approaches for in vitro data.


Histamine intolerance: lack of reproducibility of single symptoms by oral provocation with histamine: a randomised, double-blind, placebo-controlled cross-over study.
The term histamine intolerance stands for a range of symptoms involving various effector organs after the consumption of histamine-rich food. Our intention was to objectify and quantify histamine-associated symptoms and to analyse whether oral administration of the histamine-degrading enzyme diamine oxidase (DAO) caused a reduction of symptoms. Four Austrian centres participated. Patients suspected to be histamine intolerant were recruited. The first step consisted in the open oral provocation of these patients with 75 mg of liquid histamine. Patients who developed symptoms were tested in a randomised double blind crossover provocation protocol using histamine-containing and histamine-free tea in combination with DAO capsules or placebo. Main and secondary symptoms (strongest and weaker symptoms based on a ten-point scale) were defined, the grand total of all symptoms of the individual provocation steps was determined and changes in symptoms after administration of DAO were measured. Thirty nine patients reacted to the open histamine provocation and were enrolled in the blinded part. Here, both the main and secondary symptoms were not reproducible. Subjects reacted sometimes unexpectedly and randomly. Regarding the total symptom scores, the differences between the three treatment groups were statistically significant. The intake of DAO demonstrated a statistically significant reduction of histamine-associated symptoms compared to placebo (P = 0.014). Oral provocation with 75 mg of liquid histamine failed to reproduce histamine-associated single symptoms in many patients. One may suggest that histamine-intolerant subjects reacted with different organs on different occasions. As a consequence, reproducibility of single symptoms alone may not be appropriate to diagnose histamine-intolerance whereas a global symptom score could be more appropriate. The fact, that the intake of DAO capsules compared to placebo led to a statistically significant reduction of total symptom scores, may indirectly point in the same direction.


[Darwinism as a constraint of ecological pluralism].
In his respond to critical remarks of Mirkin (2003), the author claims that pluralism in ecology is not only its strength but also a weakness. Contemporary ecology became less pluralistic and this can be considered as good sign of maturing science. Ecological pluralism can be exemplified by the coexistence in 1920-30s of two different approaches to plant community: that of Frederic Clements in USA and that of Josias Braun-Blanquet in France. However the way to progress in this branch of ecology was paved rather by heretical ideas of Henry Gleason in USA and Ramensky in Russia (both authors independently developed non-holistic view of community as an assemblage of individualistically distributed species) than by "peaceful" coexistence of well-established schools, representatives of which tried not to interfere into argumentation of each other. Notable success in ecology of last decades was connected with several new methodologies, e.g. macroecology that concerned large scale of space and time. However Darwinism in its attempt to explain the order of nature referring to its origin remains the most universal and fruitful methodology of ecology. The success of Darwinism in ecology is understandable because this generalizing theory is based on the same universal principles that underlie the survival of any population. Ecologists and evolutionary biologists trying to understand various natural patterns actually deal with the same fundamental laws, i.e. exponential population growth, limitation of this growth by resource shortage and/or press of predators, the existence of individual variability in survival, etc.


Establishment and optimization of a hemp (Cannabis sativa L.) agroinfiltration system for gene expression and silencing studies.
Industrial hemp (Cannabis sativa L.) is a high-yielding annual crop primarily grown for fiber, seeds, and oil. Due to the phytochemical composition of hemp, there has been an increased interest in the market for nutraceuticals and dietary supplements for human health. Recent omics analysis has led to the elucidation of hemp candidate genes involved in the syntheses of specialized metabolites. However, a detailed study of these genes has not been undertaken due to the lack of a stable transformation system. We report for the first time an agroinfiltration system in hemp utilizing vacuum infiltration, which is an alternative method to stable transformation. A combination of 0.015% Silwett L-77, 5 mM ascorbic acid, and thirty second sonication followed by a 10-minute vacuum treatment resulted in the highest β-glucuronidase expression in the leaf, male and female flowers, stem, and root tissues. The phytoene desaturase gene was silenced with a transient hairpin RNA expression, resulting in an albino phenotype in the leaves and the male and female flowers. This agroinfiltration system would be useful for overexpression and silencing studies of target genes to regulate the yield of specialized metabolites in hemp.


Effect of Surface Chemistry on the Mechanisms and Governing Laws of Friction and Wear.
Recent studies have shown that interface chemistry, that is, the formation and breaking of chemical bonds across contacting interfaces, is closely related to the wear and friction behavior at the nanoscale. In reality, the dangling bond density (DBD) at contacting surfaces can vary greatly. Currently, it remains unclear how friction and wear mechanisms depend on DBDs and whether the Archard's law for wear and Amonton's law for friction are still applicable for contacting surfaces with different DBDs. In this work, we address these issues by studying the wear and friction behavior between two sliding diamond-like carbon surfaces by controlling DBDs via hydrogenation using molecular dynamics simulations. It is found that the chemical bond breaking and remaking across the contacting interface play the key role in determining the friction and wear behavior. During the sliding, a higher DBD leads to more chemical bond formations across the interface, causing stronger wear via either atom or cluster detachments. With the same DBD, a mechanism transition from an atom-by-atom to cluster detachments is observed by increasing the normal load. Remarkably, a fully saturated surface can exhibit a wearless friction. We further show that after necessary modifications, the Archard's law for wear and the Amonton's law for friction may be applicable at the nanoscale. The present work reveals insights into the effect of interface chemistry on the friction and wear, and it provides guidelines for effective antiwear design.


Comprehension of Co-Speech Gestures in Aphasic Patients: An Eye Movement Study.
Co-speech gestures are omnipresent and a crucial element of human interaction by facilitating language comprehension. However, it is unclear whether gestures also support language comprehension in aphasic patients. Using visual exploration behavior analysis, the present study aimed to investigate the influence of congruence between speech and co-speech gestures on comprehension in terms of accuracy in a decision task. Twenty aphasic patients and 30 healthy controls watched videos in which speech was either combined with meaningless (baseline condition), congruent, or incongruent gestures. Comprehension was assessed with a decision task, while remote eye-tracking allowed analysis of visual exploration. In aphasic patients, the incongruent condition resulted in a significant decrease of accuracy, while the congruent condition led to a significant increase in accuracy compared to baseline accuracy. In the control group, the incongruent condition resulted in a decrease in accuracy, while the congruent condition did not significantly increase the accuracy. Visual exploration analysis showed that patients fixated significantly less on the face and tended to fixate more on the gesturing hands compared to controls. Co-speech gestures play an important role for aphasic patients as they modulate comprehension. Incongruent gestures evoke significant interference and deteriorate patients' comprehension. In contrast, congruent gestures enhance comprehension in aphasic patients, which might be valuable for clinical and therapeutic purposes.


The effect of calving in the summer on the hepatic transcriptome of Holstein cows during the peripartal period.
The liver is the main metabolic organ coordinating the adaptations that take place during the peripartal period of dairy cows. A successful transition into lactation, rather than management practices alone, depends on environmental factors such as temperature, season of parturition, and photoperiod. Therefore, we analyzed the effect of calving season on the hepatic transcriptome of dairy cows during the transition period. A total of 12 Holstein dairy cows were assigned into 2 groups based on calving season (6 cows March-April, spring; 6 cows June-July, summer, SU). The RNA was extracted from liver samples obtained at -30, 3, and 35 DIM via percutaneous biopsy and hybridized to the Agilent 44K Bovine (V2) Gene Expression Microarray (Agilent Technologies Inc., Santa Clara, CA). A quantitative PCR on 22 target genes was performed to verify and expand the analyses. A total of 4,307 differentially expressed genes were detected (false discovery rate ≤0.05) in SU compared with spring. Furthermore, 73 unique differentially expressed genes were detected in SU compared with spring cows after applying a fold-change threshold ≥3 and ≤-3. For Kyoto Encyclopedia of Genes and Genomes pathways analysis of differentially expressed genes, we used the dynamic impact approach. Ingenuity Pathway Analysis software was used to analyze upstream transcription regulators and perform gene network analysis. Among metabolic pathways, energy metabolism from lipids, carbohydrates, and amino acids was strongly affected by calving in SU, with a reduced level of fatty acid synthesis, oxidation, re-esterification, and synthesis of lipoproteins, leading to hepatic lipidosis. Glycan-synthesis was downregulated in SU cows probably as a mechanism to counteract the progression of this lipidosis. In contrast, calving in the SU resulted in upregulation of gluconeogenesis but also greater use of glucose as an energy source. Among nonmetabolic pathways, the heat-shock response was obviously activated in SU cows but was also associated with inflammatory and intracellular stress response. Furthermore, data support a recent finding that cows experience endoplasmic reticulum stress around parturition. Transcription regulator analysis revealed how metabolic changes are related to important regulatory mechanisms, including epigenetic modification. The holistic analyses of the liver transcriptome response to calving in the summer at high environmental temperatures underscore how transition cows should be carefully managed during this period, as they experience alterations in liver energy metabolism and inflammatory state increasing susceptibility to health disorders in early postpartum.


Bone Infarct-Associated Osteosarcoma: Epidemiologic and Survival Trends.
Bone tumors are not a frequent occurrence and bone infarct-associated sarcomas are even rarer. The prognosis of patients experiencing this disease is poor and treatment for them remains a challenge. Nevertheless, hardly any analyses in literature report on secondary osteosarcoma (SO) on bone infarct and most of the data available do not provide sufficient details. We evaluated whether this condition could be further characterized and if prognosis could be influenced by the chemotherapy (ChT) treatment. We sought to determine: (1) the main features of this rare disease; (2) the overall survival (OS) rate; (3) the OS rate associated to ChT treatment; and (4) the correspondence between our results and published data in terms of survival. We retrospectively reviewed patients admitted at the Rizzoli Orthopedic Institute of Bologna between 1992 and 2018 (1,465 total cases of osteosarcoma). We identified a list of 11 cases of SO on bone infarct (cohort 1). We conferred about the epidemiology, surgical and ChT treatment, and surveillance of infarct-associated osteosarcoma showing the correlation to data present in literature, corresponding to 14 case reports published within 1962-2018 (cohort 2). (1) Cohort 1 was made of 11 patients: six females and five males, median age was 55 years. Nine (81%) were grade 4 and two (19%) were grade 3. Tumor predominantly arose on distal femur (64%). Most of patients had localized osteosarcoma at the diagnosis (81%); resection surgery was the elective treatment (73%) followed by amputation (18%). Of 11 patients, seven received also ChT (64%). (2) Five-year OS was 62% (95% confidence interval [CI]: 28-84). Median OS was 74 months (95% CI: 12-not reached). The cumulative incidence of cancer-related deaths (CICRD) was 37.7% (95% CI: 11.4-64.5) at 120 months. (3) In the group treated with only surgery, OS was 50% at 5 years. For patients treated with any form of ChT, OS was 71% at 5 years (p = 0.4773) and hazard ratio (HR) 0.56. The CICRD was 29% (at 2 years of follow-up. Instead, it was of 50% for patients treated only with surgery. (4) Median survival was 74 months and 12 months for cohort 1 and cohort 2, respectively (p = 0.0247). Data analysis showed a decreased HR for cohort 1 compared to cohort 2 (HR 0.315). Results confirmed also stratifying for age and ChT administration (HR 0.333). Based on this work, our opinion is that the treatment of SO patients with ChT combined to surgery improves patients' survival.


Associations between polymorphisms in the porcine haptoglobin gene and baseline levels of serum haptoglobin.
Haptoglobin (Hp) is an acute phase protein that is a marker in blood for clinical and subclinical disease in the pig. The aim of this study was to identify single nucleotide polymorphisms (SNPs) in the Hp gene and analyse their influence on baseline serum levels. DNA samples and serum were collected from 345 boars. Of 13 SNPs identified, 5 were genotyped using PCR-RFLP and Pyrosequencing. Serum Hp levels were measured using a biochemical assay. A general linear model was fitted with line and genotype as fixed effects. In addition, linkage disequilibrium (LD) was estimated between the 5 SNPs using r-square and D prime. Serum Hp concentrations in the population showed a skewed distribution with a mean of 0.34 g/L (range 0-2.65 g/L). Three SNPs were found to be associated with baseline Hp levels (p-value = 0.0093, 0.0051 and 0.0094). These 3 SNPs were also found to be in high LD with each other. This is the first study to find associations between polymorphisms in the porcine Hp gene and baseline Hp serum levels. The results have implications for breeding for resistance to infection.


Regulation of cytoplasmic pH under extreme acid conditions in suspension cultured cells of Catharanthus roseus: a possible role of inorganic phosphate.
Changes in cytoplasmic pH of suspension-cultured cells of Catharanthus roseus under extreme acid conditions were measured with the pH-dependent fluorescence dye; 2',7'-bis-(2-carboxyethyl)-5 (and-6) carboxyfluorescein (-acetoxymethylester) (BCECF). When cells were treated with 1 mM HCl (pH 3 solution), the cytoplasmic pH first decreased then returned to the original level. Treatment with 10 mM HCl (pH 2 solution) acidified the cytoplasm to a greater extent, and the acidification continued at a constant level throughout the measurement. Treatment with a pH 2 solution resulted in a gradual decrease of the malate content, indicating the operation of biochemical pH regulation mechanism. The pH 2 treatment also caused a sudden decrease of the intracellular level of Pi. The cellular content of total phosphorus did not change during the acidification. The Pi was converted to the organic phosphate form. The ATP level was not increased by the pH 2 treatment, but slightly decreased. The role of Pi, which might be functioning as a regulatory factor of cytoplasmic pH, a non-competitive inhibitor of the H+-pumps of both the plasma membrane and tonoplast is discussed.


Role of renal allograft biopsy in multicenter clinical trials in transplantation.
There is current agreement that in the design of prospective clinical trials, patients should be assigned to various treatment groups based on histological diagnosis of acute renal allograft rejection. The Banff schema is satisfactory in reporting and grading histology for patient entry into clinical trials, particularly in multicenter studies because it standardizes the entry and randomization criteria and maintains reproducibility of the histological Interpretation between the different centers. It should be emphasized that for biopsy specimens to be effective tools for either clinical practice or clinical trials, they need to be obtained before therapeutic intervention and handled by experienced histopathology laboratory technicians. Adequate tissue sampling is necessary for diagnosis and for proper grading of rejection. It is recommended that a minimum of two cores of renal tissue are needed to avoid underestimation of rejection grade. Banff classification considers a biopsy specimen to be adequate if it contains seven or more glomeruli and at least a single artery. Examination of serial step sections through the tissue samples is crucial to identify focal lesions of toxicity and to properly score intimal arteritis and tubulitis. In addition, fast interpretation of the biopsy by an experienced nephropathologist or a transplant pathologist within 24 hours optimizes the utilization of the biopsy-acquired information. At the present time, it is premature to decide on the role of graft histology as an endpoint in clinical trials, but posttreatment biopsies should be encouraged in large, multicenter, prospective trials because they may be our only tools to study postrejection changes. Semiquantitative biopsy reporting systems should be routinely included in clinical databases to allow for the prospective examination of structure-function relationships and long-term outcomes following acute rejection.


Lipoma of the esophagus.
A case report is presented of a 64-year-old female with a very large pedunculated lipoma in the esophagus. These lipomas are relatively uncommon and in spite of their large size may cause very few symptoms. Often they are first recognized after regurgitation of the tumor mass in the mouth which may, in some cases, lead to asphyxia and death. Because of the danger of asphyxia, therapy by excision of the tumor soon after diagnosis appears to be essential. A classification of benign esophageal tumors is given and the role of radiology in diagnosis described.


The six-minute walk test for children with cerebral palsy.
The 6-min walk test (6MWT) is increasingly being used as a measure of 'functional ability' in young people with cerebral palsy, despite a lack of published evidence that it is reliable for this population. This study aimed to determine the test-retest reliability of the 6MWT for ambulant 11-17-year-old adolescents with cerebral palsy. Adolescents with cerebral palsy were invited to participate in a single testing session. They performed the 6MWT twice under controlled conditions (standardized 10 m course layout, standardized instructions and encouragement), with a 30-min rest. Forty-one participants (response rate 36.6%) were recruited (26 males, 15 females; mean age 13.6+/-1.6 years). No significant difference was found between the 6-min walk distance in trials 1 and 2 for the whole group (448.7+/-96.9 m vs. 449.5+/-102.1 m; P=0.81) or by sex and gross motor function subsets. The test-retest intraclass correlation coefficient was 0.98. Bland-Altman analyses revealed a bias of only -0.9 m, and that in 95% of cases, the second 6-min walk distance would fall within +/-43.1 m of the first 6-min walk distance. All participants successfully managed the testing procedure, appeared to understand what was expected of them, and experienced no ill effects from the 6MWT. These results indicate that, when carried out according to American Thoracic Society guidelines, the 6MWT is a reliable test for young ambulant people with cerebral palsy.


Drosophila melanogaster mutations suppressible by the suppressor of Hairy-wing are insertions of a 7.3-kilobase mobile element.
Certain spontaneous mutations of Drosophila melanogaster are suppressed by su(Hw), the suppressor of Hairy-wing (3R-54.8). We find that mutations suppressible by su(Hw) result from insertions of a mobile element at the affected loci. The element, named gypsy, is approximately 7.3 kilobases long and includes 0.5-kilobase direct terminal repeats. It was first identified in DNA cloned from the bithorax chromosomal region of several Drosophila stocks carrying suppressible mutations of the bithorax complex. Cloned gypsy DNA was used as a probe to test for the association of gypsy with suppressible mutations at various other loci by hybridization in situ. Gypsy was found to be associated with 19 suppressible alleles at 10 different loci: yellow, Hairy-wing, scute, diminutive, cut, lozenge, forked, Beadex, hairy, and the bithorax complex. It was found with wild-type or nonsuppressible mutations at any of these loci. Gypsy DNA was also used as a probe to clone the element and adjacent unique DNA from the loci of some suppressible mutations. This confirmed the presence of the full-length element and also provided cloned DNA from the previously uncloned loci scute and cut. The suppressor of Hairy-wing is generally recessive and behaves as a null mutation. Thus, the disruption of normal gene function caused by the inserted gypsy element appears to require some product of the wild-type suppressor gene, su(Hw)+.


[No difference in effectiveness measured between treatment in a thermal bath and in an exercise bath in patients with rheumatoid arthritis].
To determine whether hydrotherapy in a thermomineral institution is superior to the same hydrotherapy in an ordinary hospital exercise-bath. Controlled therapeutic trial. The thermomineral institution at Arcen and the exercise bath at the Maasland Hospital in Sittard, the Netherlands. 46 patients with rheumatoid arthritis were treated in a by a skilled physiotherapist, according to a standardized exercise-scheme: 27 were treated in the thermomineral institution and 19 (control-group) in the hospital exercise-bath. Each patient received 12 treatments in 12 weeks. ENDPOINTS PARAMETERS: Morning stiffness, erythrocyte sedimentation rate, Ritchie index, amount of pain, answers to 11 questions concerning the activities of daily life, and psychosocial aspects of the disease. The various subjective and objective parameters were scored by the same physician. Statistically significant improvement was observed in both groups concerning morning stiffness. Other subjective parameters improved, but did not reach significance. Objective parameters did not change significantly. Between-group differences were not found. Hydrotherapy has a positive effect on some subjective but not on objective parameters in patients with rheumatoid arthritis, whether it is applied in a thermomineral institution or an ordinary hospital exercise bath.


Melting of monatomic glass with free surfaces.
Melting of monatomic glass with free surfaces has been studied by molecular dynamics simulations in models with Lennard-Jones-Gauss interatomic potential. Models have been heated up from a glassy state toward a normal liquid state. Atomic mechanism of melting has been analyzed via monitoring spatio-temporal arrangements of liquid-like atoms occurred during heating process. Liquid-like atoms are detected via the Lindemann criterion of melting. It is clear that the transition from glass into supercooled liquid of our "ordinary" glass with free surfaces exhibits a non-heterogeneous behavior, i.e., although liquid-like atoms initiate/grow mainly in the surface shell, significant amount of liquid-like atoms also initiates/grows simultaneously in the interior during heating process. We found three characteristic temperatures of melting of glass with a free surface. Temperature dependence of structure and various thermodynamic quantities of the system upon heating is also presented and discussed.


Endothelin-1 serum levels in women with Rheumatoid Arthritis.
The purpose of this study was to evaluate serum Endothelin-1(ET-1) levels in women Rheumatoid Arthritis (RA) patients compared with healthy controls, examine possible associations between ET-1 with different characteristic of the disease and investigate possible associations between ET-1 with surrogate markers of cardiovascular disease (CVD). This cross-sectional study was performed in Vega-Baja Hospital, Orihuela (Spain) from November 2016 to May 2018. Sixty-three women with RA and sixty-five age and sex healthy controls were included in this study. Serum ET-1 was analyzed using ELISA. Serum levels of ET-1 in RA women patients were higher than those in healthy controls (p ˂0.001). Serum levels of ET-1 were positively associated with N-terminal pro-brain natriuretic peptide (NT-proBNP) (r = 0.27, p < 0.05) and with C-reactive protein (CRP) (r = 0.36, p < 0.05). ET-1 levels in women with RA were higher in smokers. Prednisone use was associated with lower ET-1 levels. No association with carotid intima media thickness was found. we observed the presence of higher levels of serum ET-1 in RA women patients compared with healthy controls. These increased levels of ET-1 are associated with inflammation and smoking and reduced by prednisone intake.


[Comparison of multiple prediction models for hypertension (Neural networks, logistic regression and flexible discriminant analyses)].
In this study, we compared performances of logistic regression analysis (LR), flexible discriminant analysis (EAA) and neural networks (SA) in prediction of primary hypertension. Predictor variables were family history, lipoprotein A, triglyceride, smoking and body mass index. The data were collected from Cardiology Clinic of Trakya University Medical Faculty in Turkey, 2001. Logistic regression analysis, flexible discriminant analysis and neural networks were used for prediction of control and hypertension groups. Comparison of the performance of all models was done using receiver operating characteristic (ROC) curve analysis. All models had areas under the ROC curve in the range of 0.793-0.984 and SA had sensitivity, specificity, and accuracy greater than 90% at ideal threshold. ROC curve areas of SA and LR, and SA and EAA were statistically different (p<0.001 and p<0.001 respectively), while ROC curve areas of EAA and LR did not differ (p>0.05). We concluded that family history, lipoprotein A, triglyceride, smoking and body mass index variables can be used for prediction of control and hypertension groups with statistically better performance of SA over LR and EAA.


Improvement in a post-stroke pediatric patient with hemiplegia: Use of a hand-arm bimanual intensive therapy with hybrid assistive limb.
Pediatric stroke is a rare medical condition that often leads to long-lasting motor and cognitive impairments. Although therapies for adults after a stroke are well described, treatments for motor deficits following a pediatric stroke are yet to be investigated. We report a case of pediatric stroke in the chronic phase, in which a combination of novel treatments resulted in a significant improvement in physical function. A seven-year-old girl with a left hemispheric cerebral infarction lost almost all right upper extremity motor function. Following onabotulinumtoxinA treatment, she underwent hand-arm bimanual intensive therapy augmented with a hybrid assistive limb for 90 h over 15 days. Evaluation after the training revealed significant improvements in physical function, daily activities, and occupational performance. This report highlights the importance of innovative combinations of techniques in the treatment of pediatric stroke.


[QTU pattern in a patient with the Anderson-Tawil syndrome].
The Andersen-Tawil syndrome (ATS) is an autosomal dominant or sporadic disorder characterised by periodic paralysis, cardiac arrhythmias and dysmorphic features. Mutations in KCNJ2, which encodes the inward rectifier K+ channel Kir2.1 can be found in approximately 70% of ATS cases. The genetic mechanism in the remaining 30% of patients is still unknown. We present ECG of a 19-year-old man with the Andersen-Tawil syndrome. The standard ECG revealed prominent U wave with the ATS TU pattern.


Zingerone Promotes Osteoblast Differentiation Via MiR-200c-3p/smad7 Regulatory Axis in Human Bone Mesenchymal Stem Cells.
BACKGROUND Osteoblast differentiation is a critical process to maintain the stability of the bone homeostasis. Zingerone, 4-(4-hydroxy-3-methoxyphenyl)-2-butanone (ZG), isolated from ginger, performs a wide range of biological functions in human diseases. The objective of this paper was to clarify the role of ZG in human bone mesenchymal stem cells (hBMSCs) and associated mechanisms of ZG promoting osteoblast differentiation. MATERIAL AND METHODS The cytotoxicity of ZG was detected by MTT assay. The expression levels of miR-200c-3p, smad7, and osteoblast differentiation markers (alkaline phosphatase [ALP], osteocalcin [OC], osterix [OSX] and runt-related transcription factor 2 [RUNX2]) were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of smad7, ALP, OC, OSX, and RUNX2 were quantified by western blot analysis. The target mRNAs were predicted by bioinformatics tools TargetScan. The interaction between miR-200c-3p and smad7 was verified by luciferase reporter assay and RIP assay. RESULTS ZG was nontoxic to hBMSCs, and it accelerated osteoblast differentiation by inducing the expression of ALP, OC, OSX, and RUNX2. MiR-200c-3p was upregulated, but smad7 was downregulated in hBMSCs treated with ZG at different concentrations at different periods. Besides, miR-200c-3p positively regulated the expression of ALP, OC, OSX, and RUNX2 in ZG-induced hBMSCs. Moreover, miR-200c-3p targeted smad7 and strengthened the expression of ALP, OC, OSX, and RUNX2 in ZG-induced hBMSCs by downregulating smad7. CONCLUSIONS ZG contributed to osteoblast differentiation via miR-200c-3p/smad7 regulatory axis by promoting the expression of ALP, OC, OSX, and RUNX2 in hBMSCs.


Coupled Dictionary Learning for Multi-Contrast MRI Reconstruction.
Magnetic resonance (MR) imaging tasks often involve multiple contrasts, such as T1-weighted, T2-weighted and fluid-attenuated inversion recovery (FLAIR) data. These contrasts capture information associated with the same underlying anatomy and thus exhibit similarities in either structure level or gray level. In this paper, we propose a coupled dictionary learning based multi-contrast MRI reconstruction (CDLMRI) approach to leverage the dependency correlation between different contrasts for guided or joint reconstruction from their under-sampled k -space data. Our approach iterates between three stages: coupled dictionary learning, coupled sparse denoising, and enforcing k -space consistency. The first stage learns a set of dictionaries that not only are adaptive to the contrasts, but also capture correlations among multiple contrasts in a sparse transform domain. By capitalizing on the learned dictionaries, the second stage performs coupled sparse coding to remove the aliasing and noise in the corrupted contrasts. The third stage enforces consistency between the denoised contrasts and the measurements in the k -space domain. Numerical experiments, consisting of retrospective under-sampling of various MRI contrasts with a variety of sampling schemes, demonstrate that CDLMRI is capable of capturing structural dependencies between different contrasts. The learned priors indicate notable advantages in multi-contrast MR imaging and promising applications in quantitative MR imaging such as MR fingerprinting.


Anchoring effects in a wide binding pocket: the molecular basis of regioselectivity in engineered cytochrome P450 monooxygenase from B. megaterium.
The molecular basis of regioselectivity of cytochrome P450 monooxygenases from Bacillus megaterium (CYP102A1) with its flexible and widely opened active site is still not well understood. In the present work (-)-alpha-pinene bound complexes with two triple mutants were modeled to elucidate the contribution of the three major factors that mediate selectivity: active site shape, protein flexibility, and chemical reactivity of the substrate. For the triple mutant A74G F87V L188Q (GVQ), one stable, productive conformation of the substrate (conformation I) was identified by multiple molecular dynamics simulations. The model predicts pinene epoxide as a major product (42% pinene oxide, 23% verbenol) which is in agreement with the experimental product profile (70% pinene oxide, 20% verbenol). In contrast, for the triple mutant A74G F87G L188Q (GGQ) two stable productive substrate conformations were identified (conformations IIa and IIb), and verbenol was predicted as major product (81% verbenol, 16% myrtenol), which is in agreement with experimental results (77% verbenol, 10% myrtenol). The effect of chemical reactivity of the substrate was demonstrated by comparison of (-)-alpha-pinene to its regioisomer (-)-beta-pinene, where the product profile is shifted from 68% pinocarveol and 32% myrtanal in mutant GVQ, to 40% pinocarveol and 60% myrtanal in mutant GGQ. Our results strongly suggest a major role of residue 87 in anchoring (-)-alpha-pinene during substrate binding which provides a simple and elegant rationalization of the dynamic structure of this enzyme-substrate complex.


In situ habitat clearance rates and particle size preference of indigenous olympia oysters (Ostrea lurida) and non-native pacific oysters (Magallana gigas) in the north american pacific coast estuaries.
The Olympia oyster, Ostrea lurida, is the target of many restoration projects along estuaries on the North American Pacific coast, while the non-native Pacific oyster, Magallana gigas, dominates oyster aquaculture globally. Both species provide filtration functions that were investigated in three California bays using a whole-habitat, in situ approach, a laboratory particle selection experiment, and a regional physiological comparison. Measurements of chlorophyll α, temperature, salinity, and turbidity upstream and downstream, as well as point samples of seston total particulate matter and organic content to estimate habitat clearance rates (HCR, L hr-1 m-2) were collected. From February 2018 to June 2019, twenty-two trials were conducted across four sites. HCRs were highly variable within and among sites, ranging from site averages of -464 to 166 L hr-1 m-2, and not significantly different among sites, indicating field filtration performance of O. lurida habitat and M. gigas aquaculture is similar. Using a random forest regression, site was the most important predictor of HCR, with a variable importance score of 25.7 % (SD = 4.6 %). O. lurida and M. gigas had significantly different particle size selection preferences, likely affecting the quality of their filtration. This study's findings suggest that restoring O. lurida habitat may provide similar filtration benefits as M. gigas aquaculture, but the unique hydrodynamics and food quality of individual bays, as well as regional differences in filter feeder communities, must be considered in managing oyster habitat for filtration functions.


Surgical approach to functional tricuspid regurgitation: should we be more aggressive?
Functional tricuspid regurgitation (FTR) is an important clinical entity that is encountered frequently during the operative management of left-sided heart valve disease, particularly in the setting of mitral regurgitation. Failure to recognize the importance of FTR during mitral valve surgery may result in inferior early and late outcomes because of the progression of tricuspid regurgitation and right heart failure. In this review, attention is focused on recent literature, which increasingly supports the use of corrective tricuspid valve annuloplasty, and the growing consensus that FTR or tricuspid annular dilation should be more aggressively addressed at the time of cardiac surgery. The European Society of Cardiology and the European Association for Cardiothoracic Surgery (ESC/EACTS) 2012 guidelines recommend a more proactive approach to tricuspid regurgitation correction and highlight the shifting consensus toward more aggressive surgical therapy. Rigid annuloplasty rings should be used in favor of flexible bands or DeVega-style repairs. Preoperative determinants of clinical outcomes now include echocardiographic parameters of tricuspid valve tenting area, and associated right ventricular size and function. Despite data demonstrating inferior outcomes in the presence of residual tricuspid regurgitation after cardiac surgery, surgical repair for FTR appears to be underutilized. Increased recognition and correction of tricuspid regurgitation at the time of surgery is increasingly supported by published guidelines, although randomized prospective data are needed to bolster current recommendations.


Bayesian subset analysis.
As a means of assessing the importance of variation in treatment effect among patient subsets, we derived posterior distributions for subset-specific treatment effects. The effects are represented by combinations of terms for treatment and treatment-by-covariate interaction effects in familiar regression models. Exchange-ability among the interactions is a key assumption; thus, the results are of interest primarily in the context of examining a collection of subsets with no definite a priori distinction relative to treatment effect. Exchangeability leads to a shrinking of the posterior distributions of the interaction terms toward the natural origin of 0, offsetting the tendency of the estimated effects to disperse. The method is applied to parameter estimates from a proportional hazards regression analysis of survival data from a clinical trial, invoking the approximate multivariate normal distribution of the estimates. No subjective prior distributions are required. Vague priors are used for all of the regression coefficients except the treatment-by-covariate interactions, which are assumed to follow a normal distribution.


Spontaneous rupture of pyometra - a rare cause of peritonitis in elderly female.
The incidence of spontaneous perforations in pyometra is rare, being 0.01-0.5% in gynaecological patients. However, they are more common in postmenopausal females and their incidence is 13.6%. Here, we are reporting a rare case of peritonitis with a pneumoperitoneum in an elderly female, which was caused by a spontaneous perforation of pyometra. A 74-years old postmenopausal female with uncontrolled diabetes mellitus was admitted to surgical emergency with signs of diffuse peritonitis and she was in shock. After resuscitation, an emergency laparotomy was performed because of suspicion of a hollow viscous perforation with peritonitis. At laparotomy, about 2000ml of purulent fluid was found to be present in peritoneal cavity, while alimentary tract was intact. A rent with a diameter of 0.8 cm was found on fundus of uterus. A total abdominal hysterectomy with a bilateral salphingo oophorectomy was performed. Patient got discharged on 12(th) post-operative day without major complications. Histopathological studies revealed endometritis and myometritis, with no evidence of malignancy.


Factors associated with the use of diet and the use of exercise for prostate cancer by long-term survivors.
To assess the use of diet and the use of exercise for prostate cancer (and/or its treatments' side effects) by long-term survivors and whether such use is associated with selected socio-demographic, clinical, health-related quality-of-life (HRQOL) and psychological factors. Population-based cohort study in New South Wales, Australia of prostate cancer survivors aged <70 years at diagnosis and who returned a 10-year follow-up questionnaire. Validated instruments assessed patient's HRQOL and psychological well-being. Poisson regression was used to estimate adjusted relative proportions (RRs) of prostate cancer survivor groups who were currently eating differently ('using diet') or exercise differently ('using exercise') to help with their prostate cancer. 996 (61.0% of 1634) participants completed the 10-year questionnaire of whom 118 (11.8%; 95%CI[9.8-13.9]) were using diet and 78 (7.8%; 95%CI[6.2-9.5]) were using exercise to help with their prostate cancer. Men were more likely to use diet or use exercise for prostate cancer if they were younger (p-trend = 0.020 for diet, p-trend = 0.045 for exercise), more educated (p-trend<0.001, p-trend = 0.011), support group participants (p-nominal<0.001, p-nominal = 0.005), had higher Gleason score at diagnosis (p-trend<0.001, p-trend = 0.002) and had knowledge of cancer spread (p-nominal = 0.002, p-nominal = 0.001). Use of diet was also associated with receipt of androgen deprivation therapy (RR = 1.59; 95%CI[1.04-2.45]), a greater fear of cancer recurrence (p-trend = 0.010), cognitive avoidance (p-trend = 0.025) and greater perceived control of cancer course (p-trend = 0.014). Use of exercise was also associated with receipt of prostatectomy (RR = 2.02; 95%CI[1.12-3.63]), receipt of androgen deprivation therapy (RR = 2.20; 95%CI[1.34-3.61]) and less satisfaction with medical treatments (p-trend = 0.044). Few long-term prostate cancer survivors use diet or exercise to help with their prostate cancer. Survivors may benefit from counselling on the scientific evidence supporting healthy eating and regular exercise for improving quality-of-life and cancer-related outcomes.


Vitamin D Status at the Time of Hospitalization for Bronchiolitis and Its Association with Disease Severity.
To investigate the association between circulating 25-hydroxyvitamin D [25(OH)D] status at admission and disease severity among infants hospitalized for bronchiolitis and to determine whether the association differs by the form of 25(OH)D-total, bioavailable or free 25(OH)D. We conducted a 17-center prospective cohort study of 1016 US infants <12 months old hospitalized with bronchiolitis. Vitamin D status was defined by total 25(OH)D levels, and by calculated levels of bioavailable and free 25(OH)D. Bronchiolitis severity was defined by requirement for intensive care and hospital length-of-stay (LOS). Logistic and Poisson regression were used for unadjusted and multivariable analyses. The median age of hospitalized infants was 3.2 months (IQR 1.6-6.0). The median total 25(OH)D was 26.5 ng/mL (IQR 18.0-33.1); 298 (29%) infants had total 25(OH)D <20 ng/mL. In multivariable models, infants with total 25(OH)D <20 ng/mL had higher risk of requiring intensive care (aOR 1.72, 95% CI 1.12-2.64) and longer LOS (adjusted rate ratio 1.39, 95% CI 1.17-1.65) compared with infants with total 25(OH)D ≥30 ng/mL. Infants with the lowest tertile of bioavailable 25(OH)D, compared with those with the highest tertile, had longer LOS (adjusted rate ratio 1.32, 95% CI 1.07-1.62); admission to the intensive care unit was not statistically significant in the adjusted model (aOR 1.39, 95% CI 0.96-2.64). Free 25(OH)D level was not associated with severity of bronchiolitis in either unadjusted or adjusted models. In a large, multicenter cohort of US infants hospitalized for bronchiolitis, infants with total 25(OH)D <20 ng/mL had increased risk of intensive care and longer hospital LOS.


A polyimide pressure-contact multielectrode array for implantation along a submillimeter neural process in small animals.
We describe a microelectrode system for recordings from nerve bundles with diameters ranging from 20-200 microm. A novel polyimide structure allows for a planar microfabricated device to constrain a free neural process against several recording sites. This polyimide array contains multiple zigzag vias through which a small nerve process may be woven while remaining functionally intact in a live preparation. Our electrode construct features the benefits of nerve cuffs (evenly spaced electrodes in a polymer) and the functionality of a nerve hook (ability to connect to submillimeter processes). The device records extracellular action potentials in the red-swamp crayfish, Procambarus clarkii. Action potential propagation is monitored at several sites along a constrained nerve in this model organism's peripheral nervous system. Details of temporal accuracy and error sources in absolute conduction velocity measurements from microelectrode arrays are discussed.


Serum betatrophin levels are reduced in patients with full-blown polycystic ovary syndrome.
Betatrophin is defined as a new marker in glucose homeostasis and lipid metabolism. We aimed to investigate the role of serum betatrophin in full-blown polycystic ovary syndrome (PCOS) patients and 47-aged healthy women, 51 full-blown PCOS patients were included in this cross-sectional study. Betatrophin concentrations were significantly lower in PCOS group and displayed a positive correlation only with serum tryglyceride in control group (p < .05). A cutoff level (464.5 ng/L) was determined for betatrophin according to Receiver Operating Characteristic curve. Using this value, 64.7% of PCOS patients were classified as below the cutoff and in this group betatrophin was found to correlate negatively with fasting glucose, fasting insulin, and homeostasis model assessment of insulin resistance (p = .038, p = .020, and p = .014, respectively), and positively with total testosterone (p = .041). In the rest of PCOS cases (35.3%) who had betatrophin higher than cutoff, positive correlation was found with low-density lipoprotein cholesterol (p = .009). In conclusion, betatrophin levels are reduced in full-blown PCOS patients who had worse metabolic phenotype.


Spontaneous squamous cell carcinoma induced by the somatic inactivation of retinoblastoma and Trp53 tumor suppressors.
Squamous cell carcinomas (SCC) represent the most aggressive type of nonmelanoma skin cancer. Although little is known about the causal alterations of SCCs, in organ-transplanted patients the E7 and E6 oncogenes of human papillomavirus, targeting the p53- and pRb-dependent pathways, have been widely involved. Here, we report the functional consequences of the simultaneous elimination of Trp53 and retinoblastoma (Rb) genes in epidermis using Cre-loxP system. Loss of p53, but not pRb, produces spontaneous tumor development, indicating that p53 is the predominant tumor suppressor acting in mouse epidermis. Although the simultaneous inactivation of pRb and p53 does not aggravate the phenotype observed in Rb-deficient epidermis in terms of proliferation and/or differentiation, spontaneous SCC development is severely accelerated in doubly deficient mice. The tumors are aggressive and undifferentiated and display a hair follicle origin. Detailed analysis indicates that the acceleration is mediated by premature activation of the epidermal growth factor receptor/Akt pathway, resulting in increased proliferation in normal and dysplastic hair follicles and augmented tumor angiogenesis. The molecular characteristics of this model provide valuable tools to understand epidermal tumor formation and may ultimately contribute to the development of therapies for the treatment of aggressive squamous cancer.


From incomer to insider: The development of the TRANSPEC model - A systematic review of the factors influencing the effective rapid and early career TRANsition to a nursing SPECiality in differing contexts of practice.
Shortages in the speciality nursing workforce, both nationally and internationally are driving the need for the development of an evidence-based model to inform recruitment and retention into speciality nursing practice. This study aimed to identify the factors influencing rapid and early career transition into speciality nursing practice. A comprehensive systematic review of the literature was undertaken using a convergent qualitative synthesis design where results from qualitative, quantitative and mixed methods studies were transformed into qualitative findings. Databases included CINAHL, Medline, Scopus and PsycINFO. Search terms were: nurse, early career, rapid career, transition, specialty, and Medical Subject Heading terms included: professional development and educational, nursing, and continuing. Using validated tools, papers were independently assessed by a minimum of two reviewers. Twenty-three research articles were included. There were no randomized control trials. Through thematic analysis and matrix mapping of the results, the TRANSPEC model was developed. The model outlines three phases of transition: pre-entry, incomer and insider. There has been little focus on pre-entry with programs being designed at the incomer and insider phases. Impacting on these phases are three concepts: the self (professional and personal), the transition processes (informal and formal) and a sense of belonging. The overarching theme influencing the phases and concepts is the context of practice. Enablers and inhibitors influence successful transition and therefore impact on recruitment and retention. Each nurse's transition is influenced by time. For successful transition, the enablers and inhibitors impacting on the three concepts, phases and the context of practice need to be considered when developing any program. It is apparent that while previous studies have focused on the transition processes, such as curricula, the development of the self and a sense of belonging are also essential to successful transition. Further studies should include the pre-entry phase.


Exon skipping and duchenne muscular dystrophy therapy: selection of the most active U1 snRNA antisense able to induce dystrophin exon 51 skipping.
One promising approach for the gene therapy of Duchenne muscular dystrophy (DMD) is exon skipping. When thinking of possible intervention on human, it is very crucial to identify the most appropriate antisense sequences able to provide the highest possible skipping efficiency. In this article, we compared the exon 51 skipping activity of 10 different antisense molecules, raised against splice junctions and/or exonic splicing enhancers (ESEs), expressed as part of the U1 small nuclear RNA (snRNA). The effectiveness of each construct was tested in human DMD myoblasts carrying the deletion of exons 48-50, which can be treated with skipping of exon 51. Our results show that the highest skipping activity and dystrophin rescue is achieved upon expression of a U1 snRNA-derived antisense molecule targeting exon 51 splice sites in combination with an internal exon sequence. The efficacy of this molecule was further proven on an exon 45-50 deletion background, utilizing patient's fibroblasts transdifferentiated into myoblasts. In this system, we showed that the selected antisense was able to produce 50% skipping of exon 51.


Developmental changes in the topographical distribution of cells contributing to the lateral olfactory tract.
Developmental changes in the locations of cells in the olfactory bulb which contribute axons to the lateral olfactory tract (LOT) were investigated using retrograde transport of horseradish peroxidase (HRP). HRP was placed in the olfactory tubercle, LOT, and/or piriform cortex of golden hamsters aged 3,7,8 and 20 days. The numbers of labeled mitral and tufted cells were counted on equidistant sections through the olfactory bulb. In the younger animals (days 3 and 7), there was a distinct localization of labeled cells in the medial quadrant of the bulb. In the 3-day-old group 49% of all labeled cells were located in the medial quadrant, but by day 8 the per cent of cells in all 4 quadrants was equivalent. Between days 3 and 20, the average perimeter of the mitral cell layer doubled. There was a similar increase in the average relative total number of labeled cells per bulb. When pairs of brains from the different age groups with HRP placements of similar size and location were compared, the older brain always had more labeled cells. There was no relation of the number or distribution of labeled cells to the location of the injection. The number of labeled cells correlated positively with the size of the injection within each age group. We believe this is the first report of a developmental topographical gradient of the cells of origin of the LOT. This localization may be related to early topographical differences in functional activity which reflect a limited exposure to odors in the first week of life.


Identification of new human pregnane X receptor ligands among pesticides using a stable reporter cell system.
Pregnane X receptor (PXR, NR1I2) is activated by various chemically unrelated compounds, including environmental pollutants and drugs. We proceeded here to in vitro screening of 28 pesticides with a new reporter system that detects human pregnane X receptor (hPXR) activators. The cell line was obtained by a two-step stable transfection of cervical cancer HeLa cells. The first transfected cell line, HG5LN, contained an integrated luciferase reporter gene under the control of a GAL4 yeast transcription factor-binding site. The second cell line HGPXR was derived from HG5LN and stably expressed hPXR ligand-binding domain fused to GAL4 DNA-binding domain (DBD). The HG5LN cells were used as a control to detect nonspecific activities. Pesticides from various chemical classes were demonstrated, for the first time, to be hPXR activators: (1) herbicides: pretilachlor, metolachlor, and alachlor chloracetanilides, oxadiazon oxiconazole, and isoproturon urea; (2) fungicides: bupirimate and fenarimol pyrimidines, propiconazole, fenbuconazole, prochloraz conazoles, and imazalil triazole; and (3) insecticides: toxaphene organochlorine, permethrin pyrethroid, fipronil pyrazole, and diflubenzuron urea. Pretilachlor, metolachlor, bupirimate, and oxadiazon had an affinity for hPXR equal to or greater than the positive control rifampicin. Some of the newly identified hPXR activators were also checked for their ability to induce cytochrome P450 3A4 expression in a primary culture of human hepatocytes. HGPXR, with HG5LN as a reference, was grafted onto nude mice to assess compound bioavailability through in vivo quantification of hPXR activation. Altogether, our data indicate that HGPXR cells are an efficient tool for identifying hPXR ligands and establishing pesticides as hPXR activators.


The Development of the de Morton Mobility Index (DEMMI) in an independent sample of older acute medical patients: refinement and validation using the Rasch model (part 2).
This study describes the refinement and validation of the 17-item DEMMI in an independent sample of older acute medical patients. Instrument refinement was based on Rasch analysis and input from clinicians and researchers. The refined DEMMI was tested on 106 older general medical patients and a total of 312 mobility assessments were conducted. Based on the results of this study a further 2 items were removed and the 15 item DEMMI was adopted. The Rasch measurement properties of the DEMMI were consistent with estimates obtained from the instrument development sample. No differential item functioning was identified and an interval level scoring system was established. The DEMMI is the first mobility instrument for older people to be developed, refined and validated using the Rasch model. This study confirms that the DEMMI provides clinicians and researchers with a unidimensional instrument for measuring and monitoring changes in mobility of hospitalised older acute medical patients.


Optical antenna arrays in the visible range.
We report on experimental observations of highly collimated beams of radiation generated when a periodic sub-wavelength grating interacts with surface bound plasmon-polariton modes of a thin gold film. We find that the radiation process can be fully described in terms of interference of emission from a dipole antenna array and modeling the structure in this way enables the far-field radiation pattern to be predicted. The directionality, multiplicity and divergence of the beams can be completely described within this framework. Essential to the process are the surface plasmon excitations: these are the driving mechanism behind the beam formation, phase-coupling radiation from the periodic surface structure and thus imposing a spatial coherence. Detailed fitting of the experimental and modeled data indicates the presence of scattering events involving the interaction of two surface plasmon polariton modes.


Multiple system organ failure. The role of uncontrolled infection.
Multiple system organ failure (MSOF) remains a principal cause of death after major operative procedures and/or severe trauma. We studied multiple parameters in 553 consecutive emergency surgical patients to determine the incidence of MSOF, the predisposing factors to MSOF, and the sequelae of MSOF. Thirty-eight patients had MSOF; mortality was 74% for these patients. Evaluation of multiple factors demonstrated that (1) MSOF is primarily due to infection, (2) the temporal sequence of organ failure is lung, liver, gastric mucosa, and kidney, and (3) MSOF is the most common fatal expression of uncontrolled infection.


Fast spin-echo MR assessment of patients with poor outcome following spinal cervical surgery.
The aim of the investigation was to evaluate poor outcome following spinal and cervical surgery. A total of 146 consecutive patients operated with anterior discectomy and fusion (ADF) with the Cloward technique were investigated. Clinical notes, plain radiography, CT, and fast spin-echo (FSE) images were retrospectively evaluated. Some 30% of the patients had unsatisfactory clinical results within 12 months after surgery; 13% had initial improvement followed by deterioration of the preoperative symptoms, while 14.4% were not improved or worsened. Disc herniation and bony stenosis above, below, or at the fused level were the most common findings. In 45% of patients, surgery failed to decompress the spinal canal. In only 4 patients was no cause of remaining myelopathy and/or radiculopathy found. FSE demonstrated a large variety of pathological findings in the patients with poor clinical outcome after ADF. Postoperatively, patients with good clinical outcome had a lower incidence of pathological changes. FSE is considered the primary imaging modality for the cervical spine. However, CT is a useful complement in the axial projection to visualize bone changes.