Hemoconcentration is a poor predictor of severity in acute pancreatitis.
To determine whether the hematocrit (Hct) at admission or at 24 h after admission was associated with severe acute pancreatitis (AP), organ failure (OF), and pancreatic necrosis. A total of 336 consecutive patients with a first AP episode were studied. Etiology, Hct values at admission and at 24 h, development of severe AP according to Atlanta's criteria, pancreatic necrosis, OF and mortality were recorded. Hemoconcentration was defined as Hct level >44% for males and >40% for females. The t-test and chi2 test were used to assess the association of hemoconcentration to the severity, necrosis and OF. Diagnostic accuracy was also determined. Biliary disease was the most frequent etiology (n = 148). Mean Hct levels at admission were 41+/-6% for females and 46+/-7% for males (P<0.01). Seventy-eight (23%) patients had severe AP, and OF developed in 45 (13%) patients. According to contrast-enhanced computed tomography scan, 36% (54/150) patients showed pancreatic necrosis. Hct levels were elevated in 58% (55/96) and 61% (33/54) patients with interstitial and necrotizing pancreatitis, respectively. Neither Hct levels at admission nor hemoconcentration at 24 h were associated with the severity, necrosis or OF. Sensitivity, specificity and positive predictive values for both determinations were very low; and negative predictive values were between 61% and 86%, being the highest value for OF. Hct is not a useful marker to predict a worse outcome in acute pancreatitis. In spite of the high negative predictive value of hemoconcentration, the prognosis gain is limited due to an already high incidence of mild disease.


Constitutive expression of tdTomato protein as a cytotoxicity and proliferation marker for space radiation biology.
The radiation risk assessment for long-term space missions requires knowledge on the biological effectiveness of different space radiation components, e.g. heavy ions, on the interaction of radiation and other space environmental factors such as microgravity, and on the physical and biological dose distribution in the human body. Space experiments and ground-based experiments at heavy ion accelerators require fast and reliable test systems with an easy readout for different endpoints. In order to determine the effect of different radiation qualities on cellular proliferation and the biological depth dose distribution after heavy ion exposure, a stable human cell line expressing a novel fluorescent protein was established and characterized. tdTomato, a red fluorescent protein of the new generation with fast maturation and high fluorescence intensity, was selected as reporter of cell proliferation. Human embryonic kidney (HEK/293) cells were stably transfected with a plasmid encoding tdTomato under the control of the constitutively active cytomegalovirus (CMV) promoter (ptdTomato-N1). The stably transfected cell line was named HEK-ptdTomato-N1 8. This cytotoxicity biosensor was tested by ionizing radiation (X-rays and accelerated heavy ions) exposure. As biological endpoints, the proliferation kinetics and the cell density reached 100 h after irradiation reflected by constitutive expression of the tdTomato were investigated. Both were reduced dose-dependently after radiation exposure. Finally, the cell line was used for biological weighting of heavy ions of different linear energy transfer (LET) as space-relevant radiation quality. The relative biological effectiveness of accelerated heavy ions in reducing cellular proliferation peaked at an LET of 91 keV/μm. The results of this study demonstrate that the HEK-ptdTomato-N1 reporter cell line can be used as a fast and reliable biosensor system for detection of cytotoxic damage caused by ionizing radiation.


Infectious pathogens and risk of esophageal, gastric and duodenal cancers and ulcers in China: A case-cohort study.
Infection by certain pathogens is associated with cancer development. We conducted a case-cohort study of ~2500 incident cases of esophageal, gastric and duodenal cancer, and gastric and duodenal ulcer and a randomly selected subcohort of ~2000 individuals within the China Kadoorie Biobank study of >0.5 million adults. We used a bead-based multiplex serology assay to measure antibodies against 19 pathogens (total 43 antigens) in baseline plasma samples. Associations between pathogens and antigen-specific antibodies with risks of site-specific cancers and ulcers were assessed using Cox regression fitted using the Prentice pseudo-partial likelihood. Seroprevalence varied for different pathogens, from 0.7% for Hepatitis C virus (HCV) to 99.8% for Epstein-Barr virus (EBV) in the subcohort. Compared to participants seronegative for the corresponding pathogen, Helicobacter pylori seropositivity was associated with a higher risk of non-cardia (adjusted hazard ratio [HR] 2.73 [95% CI: 2.09-3.58]) and cardia (1.67 [1.18-2.38]) gastric cancer and duodenal ulcer (2.71 [1.79-4.08]). HCV was associated with a higher risk of duodenal cancer (6.23 [1.52-25.62]) and Hepatitis B virus was associated with higher risk of duodenal ulcer (1.46 [1.04-2.05]). There were some associations of antibodies again some herpesviruses and human papillomaviruses with risks of gastrointestinal cancers and ulcers but these should be interpreted with caution. This first study of multiple pathogens with risk of gastrointestinal cancers and ulcers demonstrated that several pathogens are associated with risks of gastrointestinal cancers and ulcers. This will inform future investigations into the role of infection in the etiology of these diseases.


Immune checkpoint molecules. Possible future therapeutic implications in autoimmune diseases.
During host immune response, an initial and sufficient activation is required to avoid infection and cancer, yet an excessive activation bears the risk of autoimmune reactivity and disease development. This fastidious balance of the immune system is regulated by co-stimulatory and co-inhibitory molecules, also known as immune checkpoints. Both excessive co-stimulation and insufficient co-inhibition can induce the activation and proliferation of autoreactive cells that may lead to the development of autoimmune diseases. During the last decade, a growing number of new immune checkpoint receptors and ligands have been discovered, providing an attractive approach to investigate their implication in the pathogenesis of autoimmune diseases and their potential role as targets for effective therapeutic interventions. In this review, we focus on the roles and underlying mechanisms of co-stimulatory and co-inhibitory receptors and other molecules that function as immune checkpoints in autoimmune diseases such as systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis, Sjögren's syndrome, type I diabetes and inflammatory bowel disease. We also summarize previous and current clinical trials targeting these checkpoint pathways in autoimmune diseases and discuss further therapeutic implications and possible risks and challenges.


Proteolytically active ADAM10 and ADAM17 carried on membrane microvesicles in human abdominal aortic aneurysms.
The intraluminal thrombus (ILT) of human abdominal aortic aneurysm (AAA) has been suggested to damage the underlying aortic wall, but previous work found scant activity of soluble proteases in the abluminal layer of the ILT, adjacent to the aneurysm. We hypothesised that transmembrane proteases carried by membrane microvesicles (MV) from dying cells remain active in the abluminal ILT. ILTs and AAA segments collected from 21 patients during surgical repair were assayed for two major transmembrane proteases, ADAM10 (a disintegrin and metalloprotease-10) and ADAM17. We also exposed cultured cells to tobacco smoke and assessed ADAM10 and ADAM17 expression and release on MVs. Immunohistochemistry showed abundant ADAM10 and ADAM17 protein in the ILT and underlying aneurysmal aorta. Domain-specific antibodies indicated both transmembrane and shed ADAM17. Importantly, ADAM10 and ADAM 17 in the abluminal ILT were enzymatically active. Electron microscopy of abluminal ILT and aortic wall showed MVs with ADAM10 and ADAM17. By flow cytometry, ADAM-positive microvesicles from abluminal ILT carried the neutrophil marker CD66, but not the platelet marker CD61. Cultured HL60 neutrophils exposed to tobacco smoke extract showed increased ADAM10 and ADAM17 content, cleavage of these molecules into active forms, and release of MVs carrying mature ADAM10 and detectable ADAM17. In conclusion, our results implicate persistent, enzymatically active ADAMs on MVs in the abluminal ILT, adjacent to the aneurysmal wall. The production of ADAM10- and ADAM17-positive MVs from smoke-exposed neutrophils provides a novel molecular mechanism for the vastly accelerated risk of AAA in smokers.


Naltrexone and coping skills therapy for alcohol dependence. A controlled study.
Ninety-seven alcohol-dependent patients were treated for 12 weeks in a double-blind, placebo-controlled study evaluating naltrexone and two manual guided psychotherapies in the treatment of alcohol dependence. Patients were randomized to receive either naltrexone or placebo and either coping skills/relapse prevention therapy or a supportive therapy designed to support the patient's own efforts at abstinence without teaching specific coping skills. Naltrexone proved superior to placebo in measures of drinking and alcohol-related problems, including abstention rates, number of drinking days, relapse, and severity of alcohol-related problems. Medication interacted with the type of psychotherapy received. The cumulative rate of abstinence was highest for patients treated with naltrexone and supportive therapy. For those patients who initiated drinking, however, patients who received naltrexone and coping skills therapy were the least likely to relapse.


Toxoplasma gondii Infections Alter GABAergic Synapses and Signaling in the Central Nervous System.
During infections with the protozoan parasite Toxoplasma gondii, gamma-aminobutyric acid (GABA) is utilized as a carbon source for parasite metabolism and also to facilitate parasite dissemination by stimulating dendritic-cell motility. The best-recognized function for GABA, however, is its role in the nervous system as an inhibitory neurotransmitter that regulates the flow and timing of excitatory neurotransmission. When this pathway is altered, seizures develop. Human toxoplasmosis patients suffer from seizures, suggesting that Toxoplasma interferes with GABA signaling in the brain. Here, we show that while excitatory glutamatergic presynaptic proteins appeared normal, infection with type II ME49 Toxoplasma tissue cysts led to global changes in the distribution of glutamic acid decarboxylase 67 (GAD67), a key enzyme that catalyzes GABA synthesis in the brain. Alterations in GAD67 staining were not due to decreased expression but rather to a change from GAD67 clustering at presynaptic termini to a more diffuse localization throughout the neuropil. Consistent with a loss of GAD67 from the synaptic terminals, Toxoplasma-infected mice develop spontaneous seizures and are more susceptible to drugs that induce seizures by antagonizing GABA receptors. Interestingly, GABAergic protein mislocalization and the response to seizure-inducing drugs were observed in mice infected with type II ME49 but not type III CEP strain parasites, indicating a role for a polymorphic parasite factor(s) in regulating GABAergic synapses. Taken together, these data support a model in which seizures and other neurological complications seen in Toxoplasma-infected individuals are due, at least in part, to changes in GABAergic signaling. Infections of the central nervous system can cause seizures. While inflammation in the brain has been proposed to initiate the onset of the seizures, relatively little is known about how inflammation impacts the structure and function of the neurons. Here we used a parasite called Toxoplasma gondii that infects the brain and showed that seizures arise due to a defect in signaling of GABA, which is the neurotransmitter primarily responsible for preventing the onset of seizures.


Epitope identification from fixed-complexity random-sequence peptide microarrays.
Antibodies play an important role in modern science and medicine. They are essential in many biological assays and have emerged as an important class of therapeutics. Unfortunately, current methods for mapping antibody epitopes require costly synthesis or enrichment steps, and no low-cost universal platform exists. In order to address this, we tested a random-sequence peptide microarray consisting of over 330,000 unique peptide sequences sampling 83% of all possible tetramers and 27% of pentamers. It is a single, unbiased platform that can be used in many different types of tests, it does not rely on informatic selection of peptides for a particular proteome, and it does not require iterative rounds of selection. In order to optimize the platform, we developed an algorithm that considers the significance of k-length peptide subsequences (k-mers) within selected peptides that come from the microarray. We tested eight monoclonal antibodies and seven infectious disease cohorts. The method correctly identified five of the eight monoclonal epitopes and identified both reported and unreported epitope candidates in the infectious disease cohorts. This algorithm could greatly enhance the utility of random-sequence peptide microarrays by enabling rapid epitope mapping and antigen identification.


Melatonin and cortisol "switches" during mania, depression, and euthymia in a drug-free bipolar patient.
Low melatonin and elevated cortisol levels have typically been reported during depression. The evidence that the converse is true during mania has been less well documented. In a single case design, repeated measures of nocturnal melatonin and cortisol were taken during mania, depression, and euthymia. Elevated levels of melatonin during mania and elevated cortisol levels during depression were the principal findings. There also did not appear to be any marked change in circadian rhythm of hormone output during the three clinical states. The implications of these findings in relation to noradrenergic dysfunction are discussed.


A novel micro- and nano-scale positioning sensor based on radio frequency resonant cavities.
In many micro- and nano-scale technological applications high sensitivity displacement sensors are needed, especially in ultraprecision metrology and manufacturing. In this work a new way of sensing displacement based on radio frequency resonant cavities is presented and experimentally demonstrated using a first laboratory prototype. The principle of operation of the new transducer is summarized and tested. Furthermore, an electronic interface that can be used together with the displacement transducer is designed and proved. It has been experimentally demonstrated that very high and linear sensitivity characteristic curves, in the range of some kHz/nm; are easily obtainable using this kind of transducer when it is combined with a laboratory network analyzer. In order to replace a network analyzer and provide a more affordable, self-contained, compact solution, an electronic interface has been designed, preserving as much as possible the excellent performance of the transducer, and turning it into a true standalone positioning sensor. The results obtained using the transducer together with a first prototype of the electronic interface built with cheap discrete elements show that positioning accuracies in the micrometer range are obtainable using this cost-effective solution. Better accuracies would also be attainable but using more involved and costly electronics interfaces.


Membrane remodeling by the PX-BAR protein SNX18 promotes autophagosome formation.
The membrane remodeling events required for autophagosome biogenesis are still poorly understood. Because PX domain proteins mediate membrane remodeling and trafficking, we conducted an imaging-based siRNA screen for autophagosome formation targeting human PX proteins. The PX-BAR protein SNX18 was identified as a positive regulator of autophagosome formation, and its Drosophila melanogaster homologue SH3PX1 was found to be required for efficient autophagosome formation in the larval fat body. We show that SNX18 is required for recruitment of Atg16L1-positive recycling endosomes to a perinuclear area and for delivery of Atg16L1- and LC3-positive membranes to autophagosome precursors. We identify a direct interaction of SNX18 with LC3 and show that the pro-autophagic activity of SNX18 depends on its membrane binding and tubulation capacity. We also show that the function of SNX18 in membrane tubulation and autophagy is negatively regulated by phosphorylation of S233. We conclude that SNX18 promotes autophagosome formation by virtue of its ability to remodel membranes and provide membrane to forming autophagosomes.


Investigation of the potential role of the germ cell complement in control of the expression of transferrin mRNA in the prepubertal and adult rat testis.
Iron is required for the normal development of germ cells during spermatogenesis. Because these cells have no direct access to systemic iron, there exists a shuttle system involving production and secretion of the iron-transporting protein transferrin by the Sertoli cells. Previous reports using cultures of immature Sertoli cells exposed to adult germ cells, or in vivo studies involving germ cell depleted adult rat testes, concluded that production of transferrin by Sertoli cells is modulated by germ cell complement. In the present study we have used in situ hybridisation with cRNA probes directed against the 5' and 3' ends of transferrin mRNA to examine the pattern of expression of transferrin in the immature and adult rat testis. Adult rats were treated with ethane dimethane sulphonate or methoxyacetic acid (MAA) to manipulate their testosterone levels or germ cell complement respectively. Initial findings obtained using the 3' probe showed a decrease in transferrin mRNA associated with round spermatid depletion. However, these data were not confirmed by in situ hybridisation when the 5' probe was used. The specificity of the probes was examined using Northern blotting and the 3' probe was found to hybridise to the germ cell transcript for hemiferrin even under conditions of high stringency. Examination of immature and pubertal rat testes by in situ hybridisation using the 5' transferrin-specific probe found that as early as 14 days of age the level of expression of transferrin mRNA was clearly different between tubules, and the mRNA appeared to be expressed in Leydig cells on and after day 31. In the adult rat testis, maximal expression of transferrin mRNA was found at stages VIII-XIV, calling into question the interpretation of the results of some previous studies showing expression of transferrin mRNA at all stages of the spermatogenic cycle. This stage-specific pattern of expression was not altered by acute germ cell depletion using MAA. However, Northern blot analysis showed a statistically significant increase in transferrin mRNA expression at 7 days after MAA treatment when pachytene spermatocytes were depleted from tubules at all stages of the spermatogenic cycle at which transferrin is normally expressed. In conclusion, we found that transferrin mRNA expression was not modulated by round spermatids as has been reported previously but that meiotic germ cells may influence expression of transferrin at specific stages of the spermatogenic cycle.


Photoaging and skin cancer: Is the inflammasome the missing link?
Photoaging and epithelial skin tumorigenesis are complex processes triggered mainly by UV radiation from chronic sun exposure. This leads to DNA damage and reactive oxygen species (ROS) production, which initiate an inflammatory response that alters cell structure and function. Changes in cell homeostasis and ROS production activate intracellular multiprotein platforms called inflammasomes. Inflammasomes nucleate around cytoplasmic receptors mainly of the NLR (nucleotide-binding domain and leucine-rich repeat) family and regulate caspase-1-dependant secretion of pro-inflammatory interleukin (IL)1β and IL18 cytokines, and an inflammatory form of death named pyroptosis. NLRP1 inflammasomes have taken centre stage in skin biology, as mutations in NLRP1 underlie the genetic etiology of dermatological diseases and increase the susceptibility to skin cancer. Targeting inflammasome(s) might be an important approach to improve skin inflammation, photoaging and reduce the risk of epithelial skin tumorigenesis. In this context, we discuss the potential implication of NLRP1 and NLRP3 inflammasomes.


Does exposure to agricultural chemicals increase the risk of prostate cancer among farmers?
Several studies suggest that farmers may be at increased risk of prostate cancer. The present analysis, based on a large population-based case-control study conducted among men in the Montreal area in the early 1980's, aim at identifying occupational chemicals which may be responsible for such increases. The original study enrolled 449 prostate cancer cases, nearly 4,000 patients with other cancers, as well as 533 population controls. Subjects were interviewed about their occupation histories, and a team of industrial hygienists assigned their past exposures using a checklist of some 300 chemicals. The present analysis was restricted to a study base of men who had worked as farmers earlier in their lives. There were a total of 49 men with prostate cancers, 127 with other cancers and 56 population controls. We created a pool of 183 controls combining the patients with cancers at sites other than the prostate and the population controls. We then estimated the odds ratio for prostate cancer associated with exposure to each of 10 agricultural chemicals, i.e., pesticides, arsenic compounds, acetic acid, gasoline engine emissions, diesel engine emissions, polycyclic aromatic hydrocarbons from petroleum, lubricating oils and greases, alkanes with >or=18 carbons, solvents, and mononuclear aromatic hydrocarbons. Based on a model adjusting for age, ethnicity, education, and respondent status, there was evidence of a two-fold excess risk of prostate cancer among farmers with substantial exposure to pesticides [odds ratio (OR)=2.3, 95% confidence interval (CI) 1.1-5.1], as compared to unexposed farmers. There was some suggestion, based on few subjects, of increased risks among farmers ever exposed to diesel engine emissions (OR=5.7, 95% CI 1.2-26.5). The results for pesticides are particularly noteworthy in the light of findings from previous studies. Suggestions of trends for elevated risks were noted with other agricultural chemicals, but these are largely novel and need further confirmation in larger samples.


Characterization of the indoor particles and their sources in an Antarctic research station.
Many studies have been carried out on the environmental impact of the research stations on the Antarctic continent. However, the assessment of indoor air quality in these confined environments has been neglected. The main objectives of this study are to investigate the granulometric distribution of the indoor particles in the different compartments of the Brazilian Antarctic Station, to examine the number and mass concentration of the indoor particles, to conduct chemical and morphological analyses of the indoor PM2.5, and to identify the possible sources of the PM. The results showed that Na, K, Cl, Fe, Zn, S and Si were the main elements detected. High levels of black carbon were recorded in the workshop, which may be associated with the use of diesel vehicles. To identify the human activities related to the indoor particle emission in the station, the size distribution of the particles in the living room was monitored for seven consecutive days, during normal station operation. It was possible to identify the influence of individual processes, such as incineration, cooking and the movement of people, upon the particle size number concentration. The indoor/outdoor (I/O) ratio for the total suspended particles (TSP), PM10, PM2.5 and PM1 measured was significantly larger than those reported for urban buildings. In general, the I/O ratio distribution for all the compartments shows peak values between 2.5 and 10 μm, which is often related to human activity, such as cleaning, personnel circulation or clothing surfaces. The maximum I/O ratio at this range varied from 12 to 60. In addition, the compartments affected by combustion processes tend to present a significant number of submicron particles.


Can nutrition counselling be more behavioural? Lessons learned from dietary management of cystic fibrosis.
Cystic fibrosis (CF) is a genetically-inherited disorder that results in energy imbalance. Undernutrition is common in children with CF and associated with poor health outcomes. To ensure optimal growth and nutrition, children with CF are recommended to consume 120-150% of the recommended daily allowance (RDA) for energy, but most studies show they typically are only able to achieve 100% of the RDA. While biological factors clearly contribute to poor dietary adherence, recent studies have documented behavioural and environmental barriers to adherence that includes parent-child interaction at mealtimes. While not 'abnormal', parent behaviours such as paying increased attention to the child in the form of coaxing, commanding and feeding when the child is engaged in behaviours incompatible with eating (food refusal, talking, leaving the meal) may serve to reinforce these child non-eating behaviours. Thus, dietary counselling alone, albeit necessary, is typically insufficient because of failure to specifically address these behavioural and environmental barriers to dietary treatment. Behavioural intervention that targets both nutrition education and behavioural management has been found to be effective in achieving an average increased energy intake of 4200 kJ (1000 kcal)/d and weight gain of 1.48 kg over 9 weeks in children with CF. This intervention utilizes self-monitoring, goal setting and shaping to structure the delivery of treatment. It also teaches parents to utilize child behaviour-management techniques to motivate children to increase their energy intake. These behavioural strategies include differential attention (praising and ignoring), contingency management and behavioural contracting. The potential application of these techniques to dietary counselling is suggested.


Preoperative cytological diagnosis of malignant mesothelioma of tunica vaginalis.
Malignant mesothelioma of tunica vaginalis testis is a rare aggressive neoplasm with ∼200 cases reported so far. A correct preoperative diagnosis of this rare tumor is difficult. Fine-needle aspiration cytology (FNAC) is a rapid and inexpensive diagnostic tool that may prove helpful in such cases. Very few cases have been diagnosed on cytology so far, and the descriptions of the cytological features have rarely been described in the literature. In this report we present the cytological features of malignant mesothelioma of tunica vaginalis testis which was diagnosed by FNAC and subsequently confirmed by histopatholoy and immunohistochemical (IHC) studies.


FAD roles in glucose catalytic oxidation studied by multiphase flow of extractive electrospray ionization (MF-EESI) mass spectrometry.
The role of the coenzyme flavin adenine dinucleotide (FAD) in the catalytic oxidation of glucose was elucidated by MS using a new extraction and ionization method. By a multiphase flow of liquid-gas, extractive electrospray ionization was achieved, and this technique (MF-EESI) decreased the salt-matrix interference effectively, avoided salt crystallizations at the capillary tip and increased ionization efficiency by a concentric-sprayed solvent. Notably, two intermediate complexes of FAD-glucose have been observed and differentiated for the first time using this MF-EESI technique. These intermediate complexes were demonstrated to be responsible for the hydride abstraction from glucose, as well as the cyclic coenzyme conversion of FAD during glucose oxidation. Online monitoring was also employed in MF-EESI, thereby providing a potential and informative tool to scrutinize enzymatic catalytic reactions.


Young women's lifestyle behaviours and their bone mineral density changes: a grounded theory analysis.
Only limited information is available on healthy young women's perspective of their own lifestyle behaviours. By lifestyle behaviours, e.g. smoking and physical activity, individuals have the possibility to influence bone mineral density (BMD). The aim of this study was to generate a theoretical model of lifestyle behaviours among young women with different BMD changes. Data were collected by interviewing with 11 women, and the material was analysed by means of the grounded theory. Two core categories were generated: (i) the respondents' outlook on life and (ii) their life situation. The respondents' outlook on life was either 'rigid' or 'relaxed'. Respondents who had a rigid outlook on life adjusted with others and had a decreased BMD while those with a relaxed outlook on life were doing things for fun and had an increased BMD level. Life situation also consisted of two categories: 'stagnation' and 'development'. Respondents in a static life situation did not pursue any active actions while in a developing life situation, the respondents were actively striving towards a goal. Four dimensions which characterized the respondents' outlook on life in relation to their life situation emerged: subordinating and enduring with a decreased BMD level or compromising and discerning with an increased BMD level. It seems as if the outlook on life has a greater influence than the acted lifestyle behaviour for bone development. Further research is needed to generalize the findings of this study and to explore the importance of the outlook on life among women of all ages.


Extensive nucleotide variability within a 370 kb sequence from the central region of the major histocompatibility complex.
The recent availability of the genomic sequence spanning the central and telomeric end of the major histocompatibility complex (MHC) has allowed a detailed study of its organisation, gene content and level of nucleotide variability. Previous analyses of nucleotide variability in the MHC have focused on the coding regions of the human leukocyte antigen (HLA) Class I and II genes. Non-coding nucleotide variability has been considered a by-product of exonic diversity. However, with the advent of genomic sequencing, the extent of non-coding nucleotide variability within the MHC has just begun to be appreciated. In this study, we compared different human haplotypes in 370 kb of sequence in the central region of the MHC to show the following: 1. unusually high levels of non-coding nucleotide variability, up to 80 times greater than elsewhere in the genome; 2. non-coding nucleotide variability greater than 1% at nucleotide sites distant to the Class I genes; 3. nucleotide variability greater than 1% maintained over regions containing highly linked loci; and 4. distinct troughs and peaks in the level of nucleotide variability. We will discuss these observations in relation to a possible role of nucleotide variability in the organisation of the MHC.


Insights into the role of elastin in vocal fold health and disease.
Elastic fibers are large, complex, and surprisingly poorly understood extracellular matrix macromolecules. The elastin fiber, generated from a single human gene--elastin, is a self assembling integral protein that endows critical mechanical proprieties to elastic tissues and organs such as the skin, lungs, and arteries. The biology of elastic fibers is complex because they have multiple components, a tightly regulated developmental deposition, a multistep hierarchical assembly, and unique biomechanical functions. Elastin is present in vocal folds, where it plays a pivotal role in the quality of phonation. This review article provides an overview of the genesis of elastin and its wide-ranging structure and function. Specific distribution within the vocal fold lamina propria across the lifespan in normal and pathological states and its contribution to vocal fold biomechanics will be examined. Elastin and elastin-derived molecules are increasingly investigated for their application in tissue engineering. The properties of various elastin-based materials will be discussed and their current and future applications evaluated. A new level of understanding of the biomechanical properties of vocal fold elastin composites and their molecular basis should lead to new strategies for elastic fiber repair and regeneration in aging and disease.


Risk Factors Underlying COVID-19 Lockdown-Induced Mental Distress.
Recent reports suggest that the COVID-19 lockdown resulted in changes in mental health, however, potential age-related changes and risk factors remain unknown. We measured COVID-19 lockdown-induced stress levels and the severity of depressive symptoms prior to and during the COVID-19 lockdown in different age groups and then searched for potential risk factors in a well-characterized general population-based sample. A total of 715 participants were tested for mental distress and related risk factors at two time-points, baseline testing prior to COVID-19 and follow-up testing during COVID-19, using a battery of validated psychological tests including the Perceived Stress Scale and the Patient Health Questionnaire. Longitudinal measurements revealed that the prevalence of moderate to high stress and the severity of depressive symptoms increased 1.4- and 5.5-fold, respectively, during the COVID-19 lockdown. This surge in mental distress was more severe in women, but was present in all age groups with the older age group exhibiting, cross-sectionally, the lowest levels of mental distress prior to and during the lockdown. Illness perception, personality characteristics such as a feeling of loneliness, and several lifestyle components were found to be associated with a significant increase in mental distress. The observed changes in mental health and the identified potential risk factors underlying these changes provide critical data justifying timely and public emergency-tailored preventive, diagnostic, and therapeutic mental health interventions, which should be integrated into future public health policies globally.


[Four cases with latex allergy followed by anaphylaxis to chestnut].
It is well known that patients with latex allergy have cross-reactions to various fruits, which is called a latex fruit syndrome. We report four cases with latex allergy followed by anaphylaxis to chestnut. They are all nurses of our hospital, who has personal history of atopic diseases. There were varieties in the methods of processing chestnut, presence of epicutaneous contact to chestnut, and clinical courses among the cases. All cases had positive skin prick test reactions while only two cases showed specific IgEs measured with AlaSTAT to chestnut. This fact suggests that we have to warn the risk of anaphylaxis even if one had not shown a serum specific IgE. We could follow the clinical courses and study specific IgEs to chestnut and latex in the two cases for more than two years. The titer of specific IgE was increased in the one, who could not avoid eating chestnut and contact to latex, while it was decreased in the other who could avoid the exposure to the antigens. Hevein is one of the panallergens among latex and related fruits. We studied specific IgEs to hevein on these four cases and 12 normal controls. The results showed that the former had significantly higher values of sIgEs to hevein compared to the latter (p < 0.05). We conclude that a patient with latex allergy has a high risk of contact urticaria or even anaphylaxis to the related fruits such as chestnut so that we recommend the patient with latex allergy to avoid them.


Effects of fibrin micromorphology on neurite growth from dorsal root ganglia cultured in three-dimensional fibrin gels.
The effect of fibrin matrix micromorphology on neurite growth was investigated by measuring the length of neurites growing in three-dimensional fibrin gels with well characterized micromorphologies. Dorsal root ganglia (DRGs) from 7-day chick embryos were entrapped and cultured in gels made from varying concentrations of fibrinogen (5-15 mg/mL) or calcium (2-10 mM). The length of growing neurites was measured with light videomicroscopy, and the number and diameter of fibrin fiber bundles were measured from scanning electron micrographs. An increase in fibrinogen concentration caused a decrease in the average fiber bundle thickness, an increase in the number of fiber bundles, and a marked decrease in neurite length. Gels made with different calcium concentrations had a similar range of variation in fibrin fiber bundle number or diameter, but these variations had little effect on neurite and associated nonneuronal cell outgrowth. These results provide insights into the process of neurite advance within fibrin and may be useful in the design of fibrin-based materials used for peripheral nerve regeneration. Furthermore, this study provides the first detailed experimental data on the micromorphology of fibrin matrices made from more than 5 mg/mL of fibrinogen and indicates that existing kinetic models of fibrin polymerization do not accurately predict fibrin structure at these higher concentrations.


New Tricks for Old Proteins: Single Mutations in a Nonenzymatic Protein Give Rise to Various Enzymatic Activities.
Design of a new catalytic function in proteins, apart from its inherent practical value, is important for fundamental understanding of enzymatic activity. Using a computationally inexpensive, minimalistic approach that focuses on introducing a single highly reactive residue into proteins to achieve catalysis we converted a 74-residue-long C-terminal domain of calmodulin into an efficient esterase. The catalytic efficiency of the resulting stereoselective, allosterically regulated catalyst, nicknamed AlleyCatE, is higher than that of any previously reported de novo designed esterases. The simplicity of our design protocol should complement and expand the capabilities of current state-of-art approaches to protein design. These results show that even a small nonenzymatic protein can efficiently attain catalytic activities in various reactions (Kemp elimination, ester hydrolysis, retroaldol reaction) as a result of a single mutation. In other words, proteins can be just one mutation away from becoming entry points for subsequent evolution.


Human adenovirus serotypes 4 and 11 show higher binding affinity and infectivity for endothelial and carcinoma cell lines than serotype 5.
Adenoviruses are promising vectors for human cancer gene therapy. However, the extensively used adenoviruses serotypes 2 and 5 (Ad2 and Ad5) from species C have a major disadvantage in being highly prevalent; thus, most adults have an immunity against the two viruses. Furthermore, the expression of coxsackievirus and adenovirus receptors for Ad2 and Ad5 varies in different cells. This study aims to identify adenovirus serotypes with specific tropism for endothelial cells and epithelial tumour cells. Comparison of the binding affinities of Ad31, Ad11, Ad5, Ad37, Ad4 and Ad41, belonging to species A-F, respectively, to established cell lines of hepatoma (HepG2), breast cancer (CAMA and MG7), prostatic cancer (DU145 and LNCaP) and laryngeal cancer (Hep2), as well as to endothelial cells (HMEC), was carried out by flow cytometric analysis. Ad11 from species B showed markedly higher binding affinity than Ad5 for the endothelial cell line and all carcinoma cell lines studied. Ad4 showed a specific binding affinity for hepatoma cells and laryneal carcinoma cells. The ability of Ad11, Ad4 and Ad5 to be expressed in hepatoma, breast cancer and endothelial cell lines was studied by immunostaining and (35)S-labelling of viral proteins in infected cells. Ad11 and Ad4 manifested a higher proportion of infected cells and a higher degree of hexon expression than Ad5.


Kinematic Analysis of Four Plyometric Push-Up Variations.
Plyometric research in the upper extremity is limited, with the effects of open-chain plyometric exercises being studied most. Kinematic and ground reaction force data concerning closed-chain upper extremity plyometrics has yet to be examined. Twenty-one recreationally active male subjects performed four variations of plyometric push-ups in a counterbalanced order. These included box drop push-ups from 3.8 cm, 7.6 cm, 11.4 cm heights, and clap push-ups. Kinematics of the trunk, dominant extremity and both hands were collected to examine peak flight, elbow flexion at ground contact, elbow displacement, and hand separation. Additionally peak vertical ground reaction force was measured under the dominant extremity. The 11.4 cm and clap push-ups had significantly higher peak flight than the other variations (P<.001). At ground contact, the elbow was in significantly greater flexion for the 3.8 cm and clap push-up compared to the other variations (P<.001). The clap push-up had significantly more elbow displacement than the other variations (P<.001) while hand separation was not significantly different between variations (P=.129). Peak vertical ground reaction force was significantly greater for the clap push-ups than for all other variations (P< .001). Despite similar flight heights between the 11.4 cm and clap push-ups, the greater peak vertical ground reaction force and elbow displacement of the clap push-ups indicates the clap push-up is the most intense of the variations examined. Understanding the kinematic variables involved will aid in the creation of a closed chain upper-extremity plyometric progression.


Typhoid in Laos: An 18-Year Perspective.
Although typhoid is endemic to Southeast Asia, very little is known about the disease in Laos. Typhoid vaccination is not included in the national immunization program. Although sanitation has improved, one million people still do not have access to basic clean water sources. We describe the epidemiology and antimicrobial susceptibility patterns of Salmonella enterica serovar Typhi (S. Typhi) infection in Laos based on isolates accrued over 18 years at Mahosot Hospital, Vientiane. All blood cultures collected from patients presenting with fever submitted to the Microbiology Laboratory at Mahosot Hospital (February 2000-December 2018) were included. This included patients from Vientiane and four provincial hospitals and one typhoid outbreak investigation. A total of 913 (1.5%) of 60,384 blood cultures were positive for S. Typhi. The majority of isolates with data available (712/898, 79.3%) were susceptible to all antibiotics tested, with 59 (6.5%) multidrug-resistant (MDR) isolates, mostly from one outbreak. Of 854 isolates, 12 (1.4%) were fluoroquinolone resistant. Patient admissions peaked between March and June at the end of the dry season. Although there are key limitations, these data give the first detailed epidemiological evidence of typhoid in Laos. However, estimates will be greatly influenced by access to blood culture services and health-seeking behavior. Although typhoid multidrug resistance and fluoroquinolone resistance are not currently major issues in Laos, continued surveillance and improved antibiotic stewardship are necessary to forestall worsening of the situation. Cost-effectiveness analysis is needed to inform decisions regarding typhoid vaccine introduction.


Persistent expression of Ia antigen and viral genome in visna-maedi virus-induced inflammatory cells. Possible role of lentivirus-induced interferon.
In this study we investigated the pathogenesis of the lymphoproliferative response in the chronic-active visna maedi virus-induced inflammatory lesions. Viral RNA expression was confined to macrophages, but only in tissues showing inflammatory lesions. A persistent and high level of Ia antigen expression was seen in macrophage-like cells in the inflammatory lesions, and the amounts of viral RNA and Ia expression were closely correlated. A small subpopulation of macrophages contained both viral RNA and Ia antigen, and these were found in greatest number in the lung. In vitro experiments showed that a lentivirus-induced interferon (LV-IFN) could induce Ia antigens in normal sheep spleen and lymph node cells as well as in a transformed sheep macrophage cell line. Ia antigen expression in macrophages was transient in the absence of a continuing IFN stimulus and persisted for at least 2 wk in the presence of LV-IFN. LV-IFN also restricted viral replication in macrophages. It is suggested that LV-IFN induced by the inflammatory cells in visna-maedi lesions may induce Ia antigen expression in macrophages, thereby indirectly causing the lymphoproliferative response and restricted virus replication.


Validity of the National Death Index to ascertain the date and cause of death in men having undergone prostatectomy for prostate cancer.
The National Death Index (NDI) is a centralized database containing information from death certificates that are frequently referenced by health and medical investigators to ascertain vital statistics. Yet, it commonly includes misclassified causes of death. Since the NDI is frequently relied upon in studies that evaluate outcomes following radical prostatectomy (RP) for prostate cancer (PC), we evaluated its validity by referencing mortality data from the Shared Equal Access Regional Cancer Hospital (SEARCH) database which is a prospectively managed database of 5009 Veterans who underwent a RP at eight Veterans Affairs medical centers between 1982 and 2016. We compared vital status, cause of death and date of death from the SEARCH database with the NDI. A total of 1312 men in SEARCH were deceased, yet the NDI reported 17% (219) of those men as still alive. Among the 1093 men who had concordant vital status in both SEARCH and NDI, the date of death was an exact match within one day, a week, or 31 days in 94%, 97%, 99%, and 100%, respectively. Of those men coded as dying from prostate cancer in the SEARCH database (n = 105), 12% were coded as having died from non-PC causes in the NDI. Meanwhile, among patients coded by the NDI as having died of PC (n = 139), 34% were coded in SEARCH as having died of non-PC causes. These findings demonstrate that the NDI provides accurate dates of death, but frequently misclassifies whether a death was due to prostate cancer. Studies that rely upon death certificates, as captured in the NDI, may be unreliable to report prostate cancer-specific mortality rates after prostatectomy.


Evidence of self-report bias in assessing adherence to guidelines.
To assess trends in the use of self-report measures in research on adherence to practice guidelines since 1980, and to determine the impact of response bias on the validity of self-reports as measures of quality of care. We conducted a MEDLINE search using defined search terms for the period 1980 to 1996. Included studies evaluated the adherence of clinicians to practice guidelines, official policies, or other evidence-based recommendations. Among studies containing both self-report (e.g. interviews) and objective measures of adherence (e.g. medical records), we compared self-reported and objective adherence rates (measured as per cent adherence). Evidence of response bias was defined as self-reported adherence significantly exceeding the objective measure at the 5% level. We identified 326 studies of guideline adherence. The use of self-report measures of adherence increased from 18% of studies in 1980 to 41% of studies in 1985. Of the 10 studies that used both self-report and objective measures, eight supported the existence of response bias in all self-reported measures. In 87% of 37 comparisons, self-reported adherence rates exceeded the objective rates, resulting in a median over-estimation of adherence of 27% (absolute difference). Although self-reports may provide information regarding clinicians' knowledge of guideline recommendations, they are subject to bias and should not be used as the sole measure of guideline adherence.


Silver jewelry microanalysis with dual-pulse laser-induced breakdown spectroscopy: 266 + 1064 nm wavelength combination.
Orthogonal dual-wavelength dual-pulse laser-induced breakdown spectroscopy (ODWDP-LIBS) with 266+1064 nm wavelength combination was applied to realize silver jewelry microanalysis with enhanced sensitivity and minimal sample ablation. In this technique, the 266 nm laser with low pulse energy was selected as ablation laser and the time-delayed 1064 nm laser with moderate pulse energy was selected as reheating laser to enhance plasma emission. Significant signal enhancement was achieved under the excitation of the reheating laser without increasing mass ablation which was only determined by the ablation laser. Internal standard method was applied to realize quantitative analysis of copper impurity in silver jewelry samples. The calibration curve was built, and the limit of detection of copper in silver matrix was determined to be 37.4 ppm when the crater diameter was controlled at 6.5 μm. This technique is especially useful for microanalysis of precious samples due to the property of less sample ablation in comparison with single-pulse laser-induced breakdown spectroscopy (SP-LIBS) under the same analytical sensitivity.


Deep brain stimulation can suppress pathological synchronisation in parkinsonian patients.
Although deep brain stimulation (DBS) of the subthalamic nucleus (STN) is a highly effective therapeutic intervention in severe Parkinson's disease, its mechanism of action remains unclear. One possibility is that DBS suppresses local pathologically synchronised oscillatory activity. To explore this, the authors recorded from DBS electrodes implanted in the STN of 16 patients with Parkinson's disease during simultaneous stimulation (pulse width 60 μs; frequency 130 Hz) of the same target using a specially designed amplifier. The authors analysed data from 25 sides. The authors found that DBS progressively suppressed peaks in local field potential activity at frequencies between 11 and 30 Hz as voltage was increased beyond a stimulation threshold of 1.5 V. Median peak power had fallen to 54% of baseline values by a stimulation intensity of 3.0 V. The findings suggest that DBS can suppress pathological 11-30 Hz activity in the vicinity of stimulation in patients with Parkinson's disease. This suppression occurs at stimulation voltages that are clinically effective.


Sarpogrelate, a selective 5-HT2A serotonergic receptor antagonist, inhibits serotonin-induced coronary artery spasm in a porcine model.
Serotonin is one of the most important vasoactive substances and has been implicated in the pathogenesis of coronary artery spasm and of acute coronary syndrome. We have recently demonstrated that local and long-term treatment with interleukin-1beta(IL-1beta) causes coronary arteriosclerotic changes and hyperconstrictive responses to serotonin in pigs in vivo. However, it remains to be examined which serotonergic (5-HT) receptor subtype mediates coronary spasm and whether alterations in serotonergic receptors are involved in the abnormality. In this study, we examined the inhibitory effect of sarpogrelate, a selective 5-HT2A serotonergic receptor antagonist, on the serotonin-induced coronary spasm as well as the possible alterations of serotonergic receptors in our porcine model. A segment of the porcine coronary artery was carefully dissected and aseptically wrapped with cotton mesh absorbing IL-1beta-bound microbeads from the adventitia. Two weeks after the procedure, angiographic study was performed, followed by binding assay for 5-HT1B and 5-HT2A serotonergic receptors and reverse transcription-polymerase chain reaction (RT-PCR) analysis for mRNA of those receptors. Angiographic study showed coronary vasospastic responses to serotonin at the IL-1beta-treated site. Sarpogrelate dose-dependently inhibited the serotonin-induced coronary spasm, but it did not affect the prostaglandin F2alpha-induced vasoconstriction. Radiolabeled receptor-binding assay showed that receptor affinity or receptor number of the 5-HT1B, or 5-HT2A receptors did not differ significantly between the spastic and the control sites. Furthermore, RT-PCR analysis showed that the expression of neither 5-HT2A nor 5-HT1B receptor mRNA was significantly altered at the spastic site. These results indicate that serotonin-induced coronary spasm is mediated primarily by 5-HT2A receptor in our porcine model, although the 5-HT2A receptor was not up-regulated, suggesting that alteration in the signal-transduction pathway for vascular smooth muscle contraction beyond the 5-HT2A receptor plays a primary role in the pathogenesis of coronary spasm in our porcine model.


Prevalence of skeletal and eye malformations in frogs from north-central United States: estimations based on collections from randomly selected sites.
Skeletal malformation rates for several frog species were determined in a set of randomly selected wetlands in the north-central USA over three consecutive years. In 1998, 62 sites yielded 389 metamorphic frogs, nine (2.3%) of which had skeletal or eye malformations. A subset of the original sites was surveyed in the following 2 yr. In 1999, 1,085 metamorphic frogs were collected from 36 sites and 17 (1.6%) had skeletal or eye malformations, while in 2000, examination of 1,131 metamorphs yielded 16 (1.4%) with skeletal or eye malformations. Hindlimb malformations predominated in all three years, but other abnormalities, involving forelimb, eye, and pelvis were also found. Northern leopard frogs (Rana pipiens) constituted the majority of collected metamorphs as well as most of the malformed specimens. However, malformations were also noted in mink frogs (R. septentrionalis), wood frogs (R. sylvatica), and gray tree frogs (Hyla spp.). The malformed specimens were found in clustered sites in all three years but the cluster locations were not the same in any year. The malformation rates reported here are higher than the 0.3% rate determined for metamorphic frogs collected from similar sites in Minnesota in the 1960s, and thus, appear to represent an elevation of an earlier baseline malformation rate.


Weight change in adult life and health outcomes.
To investigate the relationship between weight change in adult life and subsequent mortality and cancer incidence in women. In 1994 to 1995, all women (age range, 42 to 81) still under general practitioner observation in the United Kingdom's Royal College of General Practitioners Oral Contraception Study (n = 12,303) were sent a health survey asking about health and lifestyle issues, including current weight and weight at age 30. The main outcome measures were 6-year all-cause mortality and cancer incidence among different weight change deciles. Cox regression was used to calculate hazard ratios that were adjusted for: social class at recruitment, BMI at age 30, and age group, parity, smoking status, and hormone replacement therapy status in 1995. Women who had been obese at age 30 were more likely to die and significantly more likely to develop cancer in the 6 years after the health survey than non-obese respondents. Women reporting weight gains between age 30 and 1995 were significantly less likely to die during the 6 years after the health survey than those with a stable weight, whereas those with weight loss did not fare any better than those in the stable-weight group. Although obesity at young age was associated with subsequent mortality and cancer incidence, weight gain over a time period of 12 to 51 years appeared to be beneficial when compared with women with stable weight over the same time period. Further research is needed to confirm or refute our findings and to allow detailed examination of potential explanations for them.


SOX2 Gene Amplification and Overexpression is Linked to HPV-positive Vulvar Carcinomas.
SOX2 (SRY-related HMG-box 2) belongs to the SOX gene family of high-mobility transcription factors indispensably involved in gene regulation in pluripotent stem cells and neural differentiation. SOX2 copy number increases have been frequently reported in various types of squamous cell cancer. To better understand the effect of SOX2 aberrations on vulvar cancer phenotype and patient prognosis, we analyzed SOX2 copy number changes using fluorescence in situ hybridization and SOX2 expression by immunohistochemistry in 55 squamous cell carcinomas of the vulva. SOX2 amplification was found in 20.8% of tumors; 27.3% of vulvar carcinomas showed SOX2 protein overexpression. SOX2 amplification was correlated with SOX2 overexpression in our data set (P<0.01). Amplification of the SOX2 locus was associated with high tumor grade (P<0.05) and human papillomavirus (HPV) positivity (P<0.01). SOX2-amplified tumors showed more frequently a basaloid phenotype than nonamplified carcinomas. SOX2 protein overexpression was also correlated with basaloid phenotype and positive HPV status of vulvar carcinomas (P<0.05, each). SOX2 amplification and expression were not associated with patient overall survival. In conclusion, SOX2 copy number increases are detectable in a substantial proportion of high-grade HPV-positive vulvar carcinomas with basaloid differentiation. Our study provides further evidence for different molecular alterations in HPV-positive and HPV-negative vulvar carcinomas.


Association of Endovascular Thrombectomy vs Medical Management With Functional and Safety Outcomes in Patients Treated Beyond 24 Hours of Last Known Well: The SELECT Late Study.
The role of endovascular thrombectomy is uncertain for patients presenting beyond 24 hours of the time they were last known well. To evaluate functional and safety outcomes for endovascular thrombectomy (EVT) vs medical management in patients with large-vessel occlusion beyond 24 hours of last known well. This retrospective observational cohort study enrolled patients between July 2012 and December 2021 at 17 centers across the United States, Spain, Australia, and New Zealand. Eligible patients had occlusions in the internal carotid artery or middle cerebral artery (M1 or M2 segment) and were treated with EVT or medical management beyond 24 hours of last known well. Endovascular thrombectomy or medical management (control). Primary outcome was functional independence (modified Rankin Scale score 0-2). Mortality and symptomatic intracranial hemorrhage (sICH) were safety outcomes. Propensity score (PS)-weighted multivariable logistic regression analyses were adjusted for prespecified clinical characteristics, perfusion parameters, and/or Alberta Stroke Program Early CT Score (ASPECTS) and were repeated in subsequent 1:1 PS-matched cohorts. Of 301 patients (median [IQR] age, 69 years [59-81]; 149 female), 185 patients (61%) received EVT and 116 (39%) received medical management. In adjusted analyses, EVT was associated with better functional independence (38% vs control, 10%; inverse probability treatment weighting adjusted odds ratio [IPTW aOR], 4.56; 95% CI, 2.28-9.09; P < .001) despite increased odds of sICH (10.1% for EVT vs 1.7% for control; IPTW aOR, 10.65; 95% CI, 2.19-51.69; P = .003). This association persisted after PS-based matching on (1) clinical characteristics and ASPECTS (EVT, 35%, vs control, 19%; aOR, 3.14; 95% CI, 1.02-9.72; P = .047); (2) clinical characteristics and perfusion parameters (EVT, 35%, vs control, 17%; aOR, 4.17; 95% CI, 1.15-15.17; P = .03); and (3) clinical characteristics, ASPECTS, and perfusion parameters (EVT, 45%, vs control, 21%; aOR, 4.39; 95% CI, 1.04-18.53; P = .04). Patients receiving EVT had lower odds of mortality (26%) compared with those in the control group (41%; IPTW aOR, 0.49; 95% CI, 0.27-0.89; P = .02). In this study of treatment beyond 24 hours of last known well, EVT was associated with higher odds of functional independence compared with medical management, with consistent results obtained in PS-matched subpopulations and patients with presence of mismatch, despite increased odds of sICH. Our findings support EVT feasibility in selected patients beyond 24 hours. Prospective studies are warranted for confirmation.


Strategies towards digital and semi-automated curation in RegulonDB.
Experimentally generated biological information needs to be organized and structured in order to become meaningful knowledge. However, the rate at which new information is being published makes manual curation increasingly unable to cope. Devising new curation strategies that leverage upon data mining and text analysis is, therefore, a promising avenue to help life science databases to cope with the deluge of novel information. In this article, we describe the integration of text mining technologies in the curation pipeline of the RegulonDB database, and discuss how the process can enhance the productivity of the curators. Specifically, a named entity recognition approach is used to pre-annotate terms referring to a set of domain entities which are potentially relevant for the curation process. The annotated documents are presented to the curator, who, thanks to a custom-designed interface, can select sentences containing specific types of entities, thus restricting the amount of text that needs to be inspected. Additionally, a module capable of computing semantic similarity between sentences across the entire collection of articles to be curated is being integrated in the system. We tested the module using three sets of scientific articles and six domain experts. All these improvements are gradually enabling us to obtain a high throughput curation process with the same quality as manual curation.


Preemptive use of oral ganciclovir to prevent cytomegalovirus infection in liver transplant patients: a randomized, placebo-controlled trial.
The use of postdetection antiviral treatment of cytomegalovirus (CMV) as a strategy to prevent infection and disease in solid-organ transplant patients has not been evaluated by placebo-controlled trials. We carried out such a study in 69 patients who had received liver transplants and had positive results of CMV polymerase chain reaction within 8 weeks after transplantation but did not have concomitant CMV infection or disease. These patients were randomly assigned to receive placebo or oral ganciclovir for 8 weeks. CMV infection developed in 21% and disease developed in 12% of placebo recipients (P =.022), compared with 3% and 0%, respectively, among ganciclovir recipients (P =.003). Similarly, in the placebo arm, 55% and 36% of CMV-negative patients who received organs from CMV-positive donors developed CMV infection or disease, respectively (P =.02), compared with 11% and 0% of such patients in the ganciclovir arm (P <.01). Oral ganciclovir administered on CMV detection by PCR prevents CMV infection or disease after liver transplantation.


MR defecography in patients with dyssynergic defecation: spectrum of imaging findings and diagnostic value.
We describe the spectrum of findings and the diagnostic value of MR defecography in patients referred with suspicion of dyssynergic defecation. 48 patients (34 females, 14 males; mean age 48 years) with constipation and clinically suspected dyssynergic defecation underwent MR defecography. Patients were divided into patients with dyssynergic defecation (n = 18) and constipated patients without dyssynergic defecation (control group, n = 30). MRIs were analysed for evacuation ability, time to initiate evacuation, time of evacuation, changes in the anorectal angle (ARA-change), presence of paradoxical sphincter contraction and presence of additional pelvic floor abnormalities. Sensitivity, specificity, positive and negative predictive values and accuracy for the diagnosis of dyssynergic defecation were calculated. The most frequent finding was impaired evacuation, which was seen in 100% of patients with dyssynergic defecation and in 83% of the control group, yielding a sensitivity for MR defecography for the diagnosis of dyssynergic defecation of 100% (95% confidence interval (CI) 97-100%), but a specificity of only 23% (95% CI 7-40%). A lower sensitivity (50%; 95% CI 24-76%) and a high specificity (97%; 95% CI 89-100%) were seen with abnormal ARA-change. The sensitivity of paradoxical sphincter contraction was relatively high (83%; 95% CI 63-100%). A combined analysis of abnormal ARA-change and paradoxical sphincter contraction allowed for the detection of 94% (95% CI 81-100%) of the patients with dyssynergic defecation. MR defecography detects functional and structural abnormal findings in patients with clinically suspected dyssynergic defecation. Impaired evacuation is seen in patients with functional constipation owing to other pelvic floor abnormalities than dyssynergic defecation.


Defining dimensions of research readiness: a conceptual model for primary care research networks.
Recruitment to research studies in primary care is challenging despite widespread implementation of electronic patient record (EPR) systems which potentially make it easier to identify eligible cases. Literature review and applying the learning from a European research readiness assessment tool, the TRANSFoRm International Research Readiness instrument (TIRRE), to the context of the English NHS in order to develop a model to assess a practice's research readiness. Seven dimensions of research readiness were identified: (1) Data readiness: Is there good data quality in EPR systems; (2) Record readiness: Are EPR data able to identify eligible cases and other study data; (3) Organisational readiness: Are the health system and socio-cultural environment supportive; (4) Governance readiness: Does the study meet legal and local health system regulatory compliance; (5) Study-specific readiness; (6) Business process readiness: Are business processes tilted in favour of participation: including capacity and capability to take on extra work, financial incentives as well as intangibles such as social and intellectual capital; (7) Patient readiness: Are systems in place to recruit patients and obtain informed consent? The model might enable the development of interventions to increase participation in primary care-based research and become a tool to measure the progress of practice networks towards the most advanced state of readiness.


The Lower-Extremity Functional Test and Lower-Quadrant Injury in NCAA Division III Athletes: A Descriptive and Epidemiologic Report.
The Lower-Extremity Functional Test (LEFT) has been used to assess readiness to return to sport after a lower-extremity injury. Current recommendations suggest that women should complete the LEFT in 135 s (average; range 120-150 s) and men should complete the test in 100 s (average; range 90-125 s). However, these estimates are based on limited data and may not be reflective of college athletes. Thus, additional assessment, including normative data, of the LEFT in sport populations is warranted. To examine LEFT times based on descriptive information and off-season training habits in NCAA Division III (DIII) athletes. In addition, this study prospectively examined the LEFT's ability to discriminate sport-related injury occurrence. Descriptive epidemiology. DIII university. 189 DIII college athletes (106 women, 83 men) from 15 teams. LEFT times, preseason questionnaire, and time-loss injuries during the sport season. Men completed the LEFT (105 ± 9 s) significantly faster than their female counterparts (117 ± 10 s) (P < .0001). Female athletes who reported >3-5 h/wk of plyometric training during the off-season had significantly slower LEFT scores than those who performed ≤3 h/wk of plyometric training (P = .03). The overall incidence of a lower-quadrant (LQ) time-loss injury for female athletes was 4.5/1000 athletic exposures (AEs) and 3.7/1000 AEs for male athletes. Female athletes with slower LEFT scores (≥118 s) experienced a higher rate of LQ time-loss injuries than those with faster LEFT scores (≤117 s) (P = .03). Only off-season plyometric training practices seem to affect LEFT score times among female athletes. Women with slower LEFT scores are more likely to be injured than those with faster LEFT scores. Injury rates in men were not influenced by performance on the LEFT.


Evolution of Cognitive Rehabilitation After Stroke From Traditional Techniques to Smart and Personalized Home-Based Information and Communication Technology Systems: Literature Review.
Neurological patients after stroke usually present cognitive deficits that cause dependencies in their daily living. These deficits mainly affect the performance of some of their daily activities. For that reason, stroke patients need long-term processes for their cognitive rehabilitation. Considering that classical techniques are focused on acting as guides and are dependent on help from therapists, significant efforts are being made to improve current methodologies and to use eHealth and Web-based architectures to implement information and communication technology (ICT) systems that achieve reliable, personalized, and home-based platforms to increase efficiency and level of attractiveness for patients and carers. The goal of this work was to provide an overview of the practices implemented for the assessment of stroke patients and cognitive rehabilitation. This study puts together traditional methods and the most recent personalized platforms based on ICT technologies and Internet of Things. A literature review has been distributed to a multidisciplinary team of researchers from engineering, psychology, and sport science fields. The systematic review has been focused on published scientific research, other European projects, and the most current innovative large-scale initiatives in the area. A total of 3469 results were retrieved from Web of Science, 284 studies from Journal of Medical Internet Research, and 15 European research projects from Community Research and Development Information Service from the last 15 years were reviewed for classification and selection regarding their relevance. A total of 7 relevant studies on the screening of stroke patients have been presented with 6 additional methods for the analysis of kinematics and 9 studies on the execution of goal-oriented activities. Meanwhile, the classical methods to provide cognitive rehabilitation have been classified in the 5 main techniques implemented. Finally, the review has been finalized with the selection of 8 different ICT-based approaches found in scientific-technical studies, 9 European projects funded by the European Commission that offer eHealth architectures, and other large-scale activities such as smart houses and the initiative City4Age. Stroke is one of the main causes that most negatively affect countries in the socioeconomic aspect. The design of new ICT-based systems should provide 4 main features for an efficient and personalized cognitive rehabilitation: support in the execution of complex daily tasks, automatic error detection, home-based performance, and accessibility. Only 33% of the European projects presented fulfilled those requirements at the same time. For this reason, current and future large-scale initiatives focused on eHealth and smart environments should try to solve this situation by providing more complete and sophisticated platforms.


The impact of laparoscopic cystectomy on ovarian reserve in patients with unilateral and bilateral endometriomas.
To evaluate the effects of laparoscopic cystectomy on ovarian reserve in patients with endometriomas. Prospective study. Private and university hospitals. A total of 193 patients with endometriomas undergoing laparoscopic cyctectomy. Serum levels of antimüllerian hormone (AMH), FSH, and E₂, as well as antral follicle count (AFC) were measured preoperatively and 1 week, 3 and 9 months postoperatively for AMH, and 3 months for other values. Ovarian reserve based on the comparison of AMH alterations. The secondary end points are changes in FSH, E₂, and AFC. Serum AMH level decreased significantly from the baseline (3.86 ± 3.58 ng/mL) to 1 week (1.66 ± 1.92 ng/mL), 3 months (2.06 ± 2.5 ng/mL), and 9 months (1.77 ± 1.76 ng/mL) postoperatively. Those patients with bilateral endometriomas had significantly lower levels of AMH, 1 week, 3 and 9 months after operation. Also, patients older than 38 years had lower postoperative AMH levels. The FSH levels increased significantly from baseline to 3 months postoperatively. The AFC level increased significantly from baseline to 3 months after operation. The AMH level decreased and the FSH level increased after laparoscopic cystectomy for endometriomas, especially in older patients and those with bilateral cysts.


Leukoencephalopathy with calcifications and cysts: a purely neurological disorder distinct from coats plus.
With the identification of mutations in the conserved telomere maintenance component 1 (CTC1) gene as the cause of Coats plus (CP) disease, it has become evident that leukoencephalopathy with calcifications and cysts (LCC) is a distinct genetic entity. A total of 15 patients with LCC were identified from our database of patients with intracranial calcification. The clinical and radiological features are described. The median age (range) at presentation was 10 months (range, 2 days-54 years). Of the 15 patients, 9 presented with epileptic seizures, 5 with motor abnormalities, and 1 with developmental delay. Motor abnormalities developed in 14 patients and cognitive problems in 13 patients. Dense calcification occurred in the basal ganglia, thalami, dentate nucleus, brain stem, deep gyri, deep white matter, and in a pericystic distribution. Diffuse leukoencephalopathy was present in all patients, and it was usually symmetrical involving periventricular, deep, and sometimes subcortical, regions. Cysts developed in the basal ganglia, thalamus, deep white matter, cerebellum, or brain stem. In unaffected areas, normal myelination was present. No patient demonstrated cerebral atrophy. LCC shares the neuroradiological features of CP. However, LCC is a purely neurological disorder distinguished genetically by the absence of mutations in CTC1. The molecular cause(s) of LCC has (have) not yet been determined.


Silver nanoparticles for the detection of cell surface antigens in mass cytometry.
Mass cytometry has pioneered >40-parameter single-cell analyses that allow for the characterization of complex cellular networks at unprecedented depth. Up to 135 parameters can be simultaneously detected, but limited availability of metal tags suitable for labeling of specific probes prevents optimal exploitation of the analytical capacity of mass cytometers. To this end, we here establish the application of elemental silver nanoparticles (AgNP) of different size for reporting cell surface antigens on human leukocytes in mass cytometry assays. The mass channels at 107 Da and 109 Da are uniquely occupied by silver isotopes and do not interfere with other mass cytometry reagents. Streptavidin-coated AgNP (SA-AgNP) facilitated distinct and specific detection of various antigens, such as CD8, CD244 and CD294 on peripheral blood leukocytes pre-incubated with respective biotinylated primary antibodies. Signal intensities elicited by 40 nm-sized AgNP allowed specific detection of the low abundance antigen CD25 on both, peripheral blood regulatory T cells and CD25lo CD127+ CD4+ T cells, enabling their distinct clustering in viSNE plots. SA-AgNP were of high elemental purity, showed minor background binding to cells in immunoassays, and were compatible with previously established staining protocols for PBMC and leukocytes, facilitating their use in complex mass cytometry panels. Considering the synthesis of AgNP from isotopically purified silver, the usage of AgNP extends the analytical capacity of mass cytometry panels by one, prospectively two, additional parameters, suitable for the detection of cellular targets of low abundance. © 2016 International Society for Advancement of Cytometry.


Fish consumption and cognitive function among older people in the UK: baseline data from the OPAL study.
Observational epidemiological data suggest that habitual consumption in later life of oily fish, rich in n-3 long-chain polyunsaturated fatty acids (n-3 LCPs), is associated with better cognitive function, slower rates of cognitive decline and a lower risk of dementia. In this paper we present data on baseline fish consumption and cognitive function in cognitively healthy older people randomised onto the Older People And n- 3 Long-chain polyunsaturated fatty acid (OPAL) study. In total, 867 older people were recruited to join the OPAL study from 20 general practices in England and Wales. Participants were aged 70-79 years at baseline were free of dementia and diabetes, had a Mini-Mental State Examination score of 24 or greater and did not report daily fish oil supplement consumption. Self-reported habitual fish consumption was assessed at baseline via questions on frequency and type of fish consumption. Cognitive function at baseline was assessed via validated cognitive tests assessing memory, executive function, psychomotor speed and attention, including the Californian Verbal Learning Test (CVLT), the primary outcome of the OPAL study. Reported age at leaving full time education was recorded as a measure of educational achievement and psychological health was measured using the GHQ-30 questionnaire. Unadjusted analysis revealed significant positive associations between reported fish consumption and the CVLT scores with a mean increase of approximately 0.24 words remembered for each increase in level of reported fish consumption. These associations were noticeably attenuated on adjustment for age, gender and reported age at leaving full-time education and did not remain significant on further adjustment for GHQ-30 score. Similar associations were also observed between fish consumption and the global cognitive z-score, memory score, executive function score and delay scores in unadjusted analysis with the associations again attenuated on adjustment. Baseline data from participants randomised into the OPAL study provide support for the hypothesis that higher fish consumption is associated with better cognitive function in later life. However, although in the main associations remain after adjusting for education and psychological health, the data do not allow us to rule out the possibility of residual confounding e.g. from socioeconomic status or other health behaviours. Evidence is needed from randomised clinical trials to clarify the role of n-3 LCPs in cognitive health in later life in the normal older person population.


From QASC to QASCIP: successful Australian translational scale-up and spread of a proven intervention in acute stroke using a prospective pre-test/post-test study design.
To embed an evidence-based intervention to manage FEver, hyperglycaemia (Sugar) and Swallowing (the FeSS protocols) in stroke, previously demonstrated in the Quality in Acute Stroke Care (QASC) trial to decrease 90-day death and dependency, into all stroke services in New South Wales (NSW), Australia's most populous state. Pre-test/post-test prospective study. 36 NSW stroke services. Our clinical translational initiative, the QASC Implementation Project (QASCIP), targeted stroke services to embed 3 nurse-led clinical protocols (the FeSS protocols) into routine practice. Clinical champions attended a 1-day multidisciplinary training workshop and received standardised educational resources and ongoing support. Using the National Stroke Foundation audit collection tool and processes, patient data from retrospective medical record self-reported audits for 40 consecutive patients with stroke per site pre-QASCIP (1 July 2012 to 31 December 2012) were compared with prospective self-reported data from 40 consecutive patients with stroke per site post-QASCIP (1 November 2013 to 28 February 2014). Inter-rater reliability was substantial for 10 of 12 variables. Proportion of patients receiving care according to the FeSS protocols pre-QASCIP to post-QASCIP. All 36 (100%) NSW stroke services participated, nominating 100 site champions who attended our educational workshops. The time from start of intervention to completion of post-QASCIP data collection was 8 months. All (n=36, 100%) sites provided medical record audit data for 2144 patients (n=1062 pre-QASCIP; n=1082 post-QASCIP). Pre-QASCIP to post-QASCIP, proportions of patients receiving the 3 targeted clinical behaviours increased significantly: management of fever (pre: 69%; post: 78%; p=0.003), hyperglycaemia (pre: 23%; post: 34%; p=0.0085) and swallowing (pre: 42%; post: 51%; p=0.033). We obtained unprecedented statewide scale-up and spread to all NSW stroke services of a nurse-led intervention previously proven to improve long-term patient outcomes. As clinical leaders search for strategies to improve quality of care, our initiative is replicable and feasible in other acute care settings.


Interactions between a polystyrene particle and hydrophilic and hydrophobic surfaces in aqueous solutions.
The interaction between a colloidal polystyrene particle mounted on an AFM cantilever and a hydrophilic and a hydrophobic surface in aqueous solution is investigated. Despite the apparent simplicity of these two types of systems a variety of different types of interactions are observed. The system containing the polystyrene particle and a hydrophilic surface shows DLVO-like interactions characteristic of forces between charged surfaces. However, when the surface is hydrophobized the interaction changes dramatically and shows evidence of a bridging air bubble being formed between the particle and the surface. For both sets of systems, plateaus of constant force in the force curves are obtained when the particle is retracted from the surface after being in contact. These events are interpreted as a number of individual polystyrene molecules that are bridging the polystyrene particle and the surface. The plateaus of constant force are expected for pulling a hydrophobic polymer in a bad (hydrophilic) solvent. The plateau heights are found to be of uniform spacing and independent of the type of surface, which suggests a model by which collapsed polymers are extended into the aqueous medium. This model is supported by a full stretching curve showing also the backbone elasticity and a stretching curve obtained in pentanol, where the plateau changes to a nonlinear force response, which is typical for a polymer in a good or neutral solvent. We suggest that these polymer bridges are important in particular for the interaction between polystyrene and the hydrophilic surface, where they to some extent counteract the long-range electrostatic repulsion.


Oleuropein induces apoptosis via activation of caspases and suppression of phosphatidylinositol 3-kinase/protein kinase B pathway in HepG2 human hepatoma cell line.
Oleuropein is a polyphenol, that is found in extra‑virgin olive oil. Previous studies have shown that oleuropein inhibits cell proliferation and induces apoptosis in breast cancer, colorectal cancer and thyroid cancer. The aim of the present study was to investigate the effects of oleuropein in hepatocellular carcinoma (HCC) cells. The results of Cell Counting Kit 8 and flow cytometric analysis indicated that oleuropein effectively inhibited cell viability and induced apoptosis in HepG2 human hepatoma cells in a dose‑dependent manner, through activation of the caspase pathway. Proapoptotic Bcl‑2 family members, BAX and Bcl‑2, were involved in oleuropein‑induced apoptosis. The phosphatidylinositol 3‑kinase/protein kinase B (PI3K/AKT) signaling pathway was also shown to be involved in this process. Oleuropein was demonstrated to suppress the expression of activated AKT. In addition, AKT overexpression promoted cell survival following treatment with oleuropein, while inhibition of AKT promoted cell death. Furthermore, the data demonstrated that oleuropein induces the production of reactive oxygen species (ROS) and that the function of oleuropein is, at least partially, ROS‑dependent. These results suggest that oleuropein may be a promising novel chemotherapeutic agent in hepatocellular carcinoma.


WOX4 imparts auxin responsiveness to cambium cells in Arabidopsis.
Multipotent stem cell populations, the meristems, are fundamental for the indeterminate growth of plant bodies. One of these meristems, the cambium, is responsible for extended root and stem thickening. Strikingly, although the pivotal role of the plant hormone auxin in promoting cambium activity has been known for decades, the molecular basis of auxin responsiveness on the level of cambium cells has so far been elusive. Here, we reveal that auxin-dependent cambium stimulation requires the homeobox transcription factor WOX4. In Arabidopsis thaliana inflorescence stems, 1-N-naphthylphthalamic acid-induced auxin accumulation stimulates cambium activity in the wild type but not in wox4 mutants, although basal cambium activity is not abolished. This conclusion is confirmed by the analysis of cellular markers and genome-wide transcriptional profiling, which revealed only a small overlap between WOX4-dependent and cambium-specific genes. Furthermore, the receptor-like kinase PXY is required for a stable auxin-dependent increase in WOX4 mRNA abundance and the stimulation of cambium activity, suggesting a concerted role of PXY and WOX4 in auxin-dependent cambium stimulation. Thus, in spite of large anatomical differences, our findings uncover parallels between the regulation of lateral and apical plant meristems by demonstrating the requirement for a WOX family member for auxin-dependent regulation of lateral plant growth.


Multiple faces of pain: effects of chronic pain on the brain regulation of facial expression.
Pain behaviors are shaped by social demands and learning processes, and chronic pain has been previously suggested to affect their meaning. In this study, we combined functional magnetic resonance imaging with in-scanner video recording during thermal pain stimulations and use multilevel mediation analyses to study the brain mediators of pain facial expressions and the perception of pain intensity (self-reports) in healthy individuals and patients with chronic back pain (CBP). Behavioral data showed that the relation between pain expression and pain report was disrupted in CBP. In both patients with CBP and healthy controls, brain activity varying on a trial-by-trial basis with pain facial expressions was mainly located in the primary motor cortex and completely dissociated from the pattern of brain activity varying with pain intensity ratings. Stronger activity was observed in CBP specifically during pain facial expressions in several nonmotor brain regions such as the medial prefrontal cortex, the precuneus, and the medial temporal lobe. In sharp contrast, no moderating effect of chronic pain was observed on brain activity associated with pain intensity ratings. Our results demonstrate that pain facial expressions and pain intensity ratings reflect different aspects of pain processing and support psychosocial models of pain suggesting that distinctive mechanisms are involved in the regulation of pain behaviors in chronic pain.


Human periodontal fibroblast response to enamel matrix derivative, amelogenin, and platelet-derived growth factor-BB.
The ideal goal of clinical therapy in periodontal defects is regeneration of all lost structures. For regeneration to occur, cell proliferation, migration, and extracellular matrix synthesis are prerequisites. Attempts at regeneration of periodontal defects by guided tissue regeneration using bone grafts and membranes have not always yielded predictable results. Recently, attempts at engineering the defects using various materials have shown promising results. Two such approaches have been used to regenerate periodontal defects, one using extracellular matrix such as enamel matrix proteins and the other using growth factors. However, to our knowledge, no study has looked at combining these two approaches to achieve potentially even greater regeneration. Primary human periodontal ligament (PDL) fibroblasts were explanted, and alkaline phosphatase (ALK PHOS) activity was determined. Phenotypically different cell lines were incubated for 1, 3, 6, and 10 days in 0.2% fetal bovine serum (FBS) media containing different concentrations of either enamel matrix derivative (EMD), amelogenin, platelet-derived growth factor-BB (PDGF-BB), EMD+PDGF-BB, or amelogenin+PDGF-BB. A culture of 0.2% FBS alone served as a negative control, and a culture of 10% FBS served as a positive control. Cell proliferation was measured using a Coulter counter to determine the cell number. The effects on a wound-fill model were evaluated by scraping a 3-mm wide cell-free zone in PDL monolayers across the diameter of the tissue-culture plate and determining PDL cell migration into the cell-free zone using computer assisted histomorphometry. Compared to the control, only EMD+PDGF-BB significantly increased PDL cell proliferation in an ALK PHOS (-) cell line (P<0.001), and EMD alone, EMD+PDGF-BB, and amelogenin+PDGF-BB significantly increased PDL cell proliferation in an ALK PHOS (+) cell line (P<0.001) with EMD+PDGF-BB showing a trend for greater proliferation than either PDGF or EMD alone. Individually, EMD and amelogenin had no significant effect on PDL cell proliferation. In the wound-fill experiment, all factors and their combinations except amelogenin significantly enhanced cell migration compared to the control (P<0.05) at the wound edge. In addition, EMD+PDGF-BB had additive effects on the ALK PHOS (-) cell line at the wound edge. At the center of the wound, neither EMD nor amelogenin had a significant wound-fill effect. However, the combination of EMD+PDGF-BB additively increased wound fill for both ALK PHOS (+) and ALK PHOS (-) cells. The combination of EMD and PDGF-BB produces greater proliferative and wound-fill effects on PDL cells than each by themselves. If these combined effects can be translated clinically, one may see greater regeneration in periodontal defects with this combination. However, amelogenin does not have significant effects on PDL cell proliferation or migration by itself. This may suggest that either another enamel matrix component in EMD may be responsible for some of its clinical effects, or that amelogenin alone may not trigger the regenerative potential of periodontal tissues and that it requires a combined interaction with other enamel matrix components of EMD to direct the regenerative process.


Interaction of the adenovirus L1 52/55-kilodalton protein with the IVa2 gene product during infection.
The adenovirus L1 52/55-kDa protein is expressed both in the early and late stages of infection, raising the possibility that it has multiple roles in the viral life cycle. To obtain possible insights into these roles, the yeast two-hybrid system was used to examine the interactions of the 52/55-kDa protein with viral and cellular factors. cDNA expression libraries from human 293 cells at both early and late stages of adenovirus type 5 infection were constructed and screened, with the 52/55-kDa protein being used as bait. Characterization of positive clones revealed that the adenovirus IVa2 gene product interacted specifically with the 52/55-kDa protein. In addition, the IVa2 protein was shown to interact with a bacterial glutathione S-transferase-52/55-kDa fusion protein in vitro, further supporting the finding with the yeast two-hybrid system. Finally, coimmunoprecipitation studies confirmed that the 52/55-kDa protein and IVa2 polypeptide interact specifically during the course of adenovirus infection. A potential role for the IVa2-52/55-kDa protein interaction in the regulation of transcription from the major late promoter and in viral assembly is discussed.


Evaluation of the Vicorder, a novel cuff-based device for the noninvasive estimation of central blood pressure.
The Vicorder is a new brachial cuff-based device that estimates central blood pressure (cBP) using a brachial-to-aortic transfer function. The aim of this study was to evaluate cBP estimated by the Vicorder. During cardiac angiography, cBP estimated by the Vicorder and the SphygmoCor was evaluated against simultaneous invasive cBP in 50 patients. The two devices were also compared noninvasively in a separate group of 90 healthy individuals. Central SBP (cSBP) obtained with each device satisfied the American Association for the Advancement of Medical Instrumentation accuracy criteria when peripheral waveforms were calibrated to invasive mean arterial pressure (MAP)/DBP: estimated - invasive cSBP difference, -4.0 ± 7.4 mmHg, Vicorder, P < 0.001; -1.4 ± 7.9 mmHg, SphygmoCor, P = 0.21. When oscillometric brachial SBP/DBP was used for peripheral waveform calibration, cSBP was underestimated by Vicorder (Δ -6.4 ± 7.4 mmHg, P < 0.001 versus invasive) and more so by SphygmoCor (Δ -11.9 ± 7.2 mmHg, P < 0.001 versus invasive). Conversely, cSBP was more closely estimated by SphygmoCor when waveforms were calibrated to brachial MAP/DBP (Δ -2.8 ± 9.4 mmHg, P = 0.04 versus invasive). In the noninvasive study, Vicorder cSBP correlated well with SphygmoCor cSBP when SphygmoCor waveforms were calibrated to brachial MAP/DBP (121 ± 16 versus 121 ± 17 mmHg, P = 0.2) but not when brachial SBP/DBP was used for calibration (115 ± 19 mmHg, P < 0.001). The Vicorder and SphygmoCor devices provide reliable estimates of cSBP when calibrated to invasive pressure. When calibrated to brachial BP, both devices underestimated cSBP, although this was attenuated when SphygmoCor was calibrated to brachial MAP/DBP. Vicorder may be a simple alternative to tonometry-based methods for noninvasive assessment of cBP.


Predictors of growth kinetics and outcomes in small renal masses (SRM ≤4 cm in size): Tayside Active Surveillance Cohort (TASC) Study.
To determine outcomes of small renal masses (≤4 cm) on active surveillance and explore factors which can influence their growth. Two hundred twenty six patients between January 2007 and December 2014 were analysed using cross-linked methodology of healthcare data and independent review. Cancer specific and non-specific survival were the primary outcomes. Growth kinetics, factors influencing growth and need for interventions were secondary outcomes. 101 (64.4%) solid and 4 (5.9%) cystic SRMs showed growth. 43 (19.02%) of SRMs required treatment interventions. Seven patients (7/158; 4.4%) died due to renal cancer at a median follow-up of 21.7 (SD 10.6, min 6-42) months, all in solid category. Independent review of serial radiological imaging of these seven cases showed two patients had subtle metastatic disease at the initial presentation, and 5 of the 7 did not adhere to recommended imaging regime. 33 (33/158; 20.8%) died due to other causes including non-renal cancers (14/158; 8.8%). Multivariate analyses showed that lower eGFR at baseline, co-morbidities and tumour location were independently associated with growth in size. A higher cancer-specific mortality was seen in the present study compared to the reported literature. Independent critical review of imaging of cases with poor outcome underscored the importance of adherence to a robust protocol including follow up. Comorbid conditions had a significant impact on growth and overall survival of patients with SRMs.


Osteogenic nanostructured titanium surfaces with antibacterial properties under conditions that mimic the dynamic situation in the oral cavity.
The study aim was to assess the impact of different surface nanofeatures on otherwise smooth titanium surfaces on bacterial adhesion as well as on their osteogenic potential. Bacterial adhesion was assessed in the presence of saliva under static and dynamic conditions to approximate both sub- and supragingival conditions in the oral cavity as the gingival seal will be affected by implantation. The ultimate goal was to develop a surface that will reduce biofilm formation but still support osseointegration in vivo. To this end nanotubular or nanopitted surfaces were created on electropolished titanium via electrochemical anodization procedures. Sandblasted/acid etched surfaces (SBAE) were used as a microrough reference. Bacterial adhesion was studied using saliva-precoated samples with S. sanguinis as a typical early colonizer of the oral cavity; osteogenic differentiation was assessed with human bone marrow stromal cells. While bacterial adhesion was reduced on all microsmooth surfaces to an average of 17% surface coverage compared to 61% on SBAE under static conditions, under dynamic conditions the nanopitted surface had a significant impact on bacterial adhesion. Here fluid flow removed all bacteria. By comparison, the reduction on the nanotubular surface was only similar to that of the SBAE reference. We hypothesise the underlying cause to be an effect of the surface morphology on the structure and composition of the saliva precoating that reduces its stability, giving rise to a self-cleaning effect. In addition, no negative influence on the osteogenic potential of the nanopitted surface could be determined by alkaline phosphatase activity, mineralization behaviour or gene expression; it remained on a par with the tissue culture plastic control. Thus, nanopitting seems to be a promising surface treatment candidate for dental implants to reduce infection related complications without compromising the implant integration.