Catecholamine infusion in vagotomized dogs during thiamylal-halothane and pentobarbital anesthesia.
The cardiac arrhythmogenic infusion rate of epinephrine, dopamine, and dobutamine in vagotomized dogs was determined during thiamylal-halothane and pentobarbital anesthesia. Epinephrine, dopamine, and dobutamine were administered until 4 or more ventricular arrhythmias on duplicated trials were produced or until a predetermined maximum infusion rate was attained. The mean ventricular arrhythmogenic infusion rates (micrograms X kg-1 X min-1) during thiamylal-halothane anesthesia were: epinephrine, 0.57 +/- 0.24; dopamine, 23.7 +/- 8.26; and dobutamine, 10.21 +/- 3.54. Few arrhythmias were produced at the maximum administered infusion rate during pentobarbital anesthesia (2 of 6 with epinephrine, 3 of 6 with dopamine, and 0 of 6 administered dobutamine). Heart rate and blood pressure increased progressively with increasing infusion rates for all 3 catecholamines during thiamylal-halothane anesthesia. Heart rate and blood pressure changes were similar during pentobarbital anesthesia except for blood pressure changes during dobutamine infusion.


Structural polymorphisms in fibrillar aggregates associated with exfoliation syndrome.
Exfoliation syndrome is largely considered an age-related disease that presents with fibrillar aggregates in the front part of the eye. A growing body of literature has investigated structural diversity of amyloids and fibrillar aggregates associated with neurodegenerative disease. However, in case of exfoliation syndrome, there is a dearth of information on the biophysical characteristics of these fibrils and structural polymorphisms. Herein, structural diversity of fibrils isolated from the anterior lens capsule of patients was evaluated using transmission electron microscopy techniques. It was apparent that, despite having a low sample number of different patients, there exists a wide range of fibril morphologies. As it is not precisely understood how these fibrils form, or what they are composed of, it is difficult to postulate a mechanism responsible for these differences in fibril structure. However, it is apparent that there is a wider range of fibril structure than initially appreciated. Moreover, these data may suggest the variance in fibril structure arises from patient-specific fibril composition and/or formation mechanisms.


[Catabolism of amino acids in bacteria of the genus Klebsiella].
Propagation and activity level of 18 enzymes catalyzing deamination reactions of dicarboxylic and oxyamino acids and enzymes of amino acid reamination and amino acid N-acyl-derivatives' deacylation have been studied in Klebsiella bacteria. Klebsiella the most actively utilizes serin, threonine, aspartic and glutamic acids and aromatic amino acids. The first three amino acids are utilized by deamination, aromatic acids- in aminotransferase reaction with alpha-ketoglutaric acid, glutamic acid--by deamination and decarboxylation. Besides, Klebsiella actively deacylates N-acyl-derivatives of amino acids.


Phenotype and effector function of CC chemokine receptor 9-expressing lymphocytes in small intestinal Crohn's disease.
CCL25/CCR9 chemokine ligand/receptor pair has been reported to play an important role in small bowel (SB) immunity and inflammation. We have previously reported an aberrant SB expression of CCL25 in Crohn's disease (CD) and an increased frequency of CCR9(+) T cells in the peripheral blood of patients with SB inflammatory diseases such as CD and celiac disease. In this study, we have characterized the phenotype and effector function of CCR9(+) T cells in mucosal lymphoid tissues in CD. We show that CCR9(+) T cells isolated from mesenteric lymph nodes (MLN) draining CD SB express a more activated phenotype compared with MLN draining normal SB. Stimulation of CCR9(+) T cells isolated from CD SB lamina propria produced more IFN-gamma and IL-17 in response to anti-CD3 or IL-12/IL-18 stimulation compared with those isolated from normal SB. The addition of TL1A to the cytokine combination markedly augmented the secretion of IFN-gamma, but not IL-17, by CD lamina propria CCR9(+) T cells. CCL25 incubation of CD SB lamina propria lymphocytes and MLN lymphocytes increased their adhesion to VCAM-1/Fc in vitro. Finally, the TCRVbeta analysis of CCR9(+) T cells revealed a diverse TCRVbeta repertoire among MLN CCR9(+) T cells in patients with SB CD. Our data indicate that CCR9(+) T cells in SB CD are proinflammatory and support the rationale for the use of CCR9 antagonists for the treatment of human SB CD.


How Robust Is the Ligand Binding Transition State?
For many drug targets, it has been shown that the kinetics of drug binding (e.g., on rate and off rate) is more predictive of drug efficacy than thermodynamic quantities alone. This motivates the development of predictive computational models that can be used to optimize compounds on the basis of their kinetics. The structural details underpinning these computational models are found not only in the bound state but also in the short-lived ligand binding transition states. Although transition states cannot be directly observed experimentally due to their extremely short lifetimes, recent successes have demonstrated that modeling the ligand binding transition state is possible with the help of enhanced sampling molecular dynamics methods. Previously, we generated unbinding paths for an inhibitor of soluble epoxide hydrolase (sEH) with a residence time of 11 min. Here, we computationally modeled unbinding events with the weighted ensemble method REVO (resampling of ensembles by variation optimization) for five additional inhibitors of sEH with residence times ranging from 14.25 to 31.75 min, with average prediction accuracy within an order of magnitude. The unbinding ensembles are analyzed in detail, focusing on features of the ligand binding transition state ensembles (TSEs). We find that ligands with similar bound poses can show significant differences in their ligand binding TSEs, in terms of their spatial distribution and protein-ligand interactions. However, we also find similarities across the TSEs when examining more general features such as ligand degrees of freedom. Together these findings show significant challenges for rational, kinetics-based drug design.


Multi-forms of human MTH1 polypeptides produced by alternative translation initiation and single nucleotide polymorphism.
The human MTH1 gene for 8-oxo-7,8-dihydrodeoxyguanosine triphosphatase, produces seven types (types 1, 2A, 2B, 3A, 3B, 4A and 4B) of mRNAs. The B-type mRNAs with exon 2b-2c segments have three additional in-frame AUGs in their 5' regions. We report here that these transcripts produce three forms of MTH1 polypeptides (p22, p21 and p18) in in vitro translation reactions. Three polypeptides were also detected in extracts of human cells, using western blotting. B-type mRNAs with a polymorphic alteration (GU-->GC) at the beginning of exon 2c that converts an in-frame UGA to CGA yielding another in-frame AUG further upstream, produced an additional polypeptide (p26) in vitro. Substitution of each AUG abolished the production of each corresponding polypeptide. Cell lines from individuals with the GC allele contain more B-type mRNAs than do those of GT homozygotes, and the former produce all of four polypeptides but the latter lack p26. Amounts of each polypeptide reflected copy number of the GC allele in each cell line. There is an apparent linkage dis-equilibrium between the two polymorphic sites, GT/GC at exon 2c and Val83/Met83 at codon 83 for p18.


Variation in individual responses to the cytotoxicity of cigarette smoke.
There is increasing evidence that elastase may mediate pulmonary damage in emphysema and also that the release of intracellular elastase by cigarette smoke is necessarily associated with cell death. We report consistent and significant variation in response of lymphocytes from different individuals to the cytotoxic effect of cigarette smoke condensate in vitro. This variation which occurred at levels of cigarette smoke likely to be encountered in vivo and which was independent of the age, sex, or smoking habits of the subjects, may be relevant in determining the risk of developing emphysema amongst smokers.


How representative are dissolved organic matter (DOM) extracts? A comprehensive study of sorbent selectivity for DOM isolation.
Solid phase extraction (SPE) has become a widespread method for isolating dissolved organic matter (DOM) of diverse origin such as fresh and marine waters. This study investigated the DOM extraction selectivity of 24 commercially available SPE sorbents under identical conditions (pH = 2, methanol elution) on the example of Suwannee River (SR) water and North Sea (NS) water by using DOC analysis and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Proton nuclear magnetic resonance (1H NMR) spectroscopy was employed to assess leaching behavior, and HLB sorbent was found to leach substantially, among others. Variable DOC recoveries observed for SR DOM and NS DOM were primarily caused by the respective molecular composition, with subordinated and heterogeneous contributions of relative salinity. Scatter of average H/C and O/C elemental ratios and gross alignment in mass-edited H/C ratios according to five established coarse SPE characteristics was near identical for SR DOM and NS DOM. FTMS-based principal component analysis (PCA) provided essentially analogous alignment of SR DOM and NS DOM molecular compositions according to the five established groups of SPE classification, and corroborated the sorption-mechanism-based selectivity of DOM extraction in both cases. Evaluation of structural blanks and leaching of SPE cartridges requires NMR spectroscopy because FT-ICR mass spectrometry alone will not reveal inconspicuous displacements of continual bulk signatures caused by leaching of SPE resin constituents.


Why values elicitation techniques enable people to make informed decisions about cancer trial participation.
Decision aids help patients make informed treatment decisions. Values clarification (VC) techniques are part of decision aids that help patients assimilate the information with their personal values. There is little evidence that these techniques contribute to enhanced decision making over and above the provision of good quality information. To assess whether VC techniques are active ingredients in enhancing informed decision making and explain how and why they work. Participants were randomly assigned to one of three groups: (i) information only, (ii) information plus implicit task, (iii) information plus explicit task. Thirty healthy women from a UK University participated by making a hypothetical choice between taking part in a clinical trial and having the standard treatment for breast cancer. Verbal protocols were elicited by think-aloud method and content analysed to assess informed decision making; a questionnaire was completed after the decision assessing decision preference, perceptions of decisional conflict and ambivalence. Data were analysed using multivariate statistics. No participants changed their decision preference as a result of the VC techniques. Women in the explicit VC group evaluated more information in accord with personal values, expressed lower ambivalence, decisional uncertainty and greater clarity of personal values than those in the implicit VC and control groups. Feelings of ambivalence about both options were related to decisional conflict. Explicit VC techniques are likely to be active ingredients in decision aids. They work by enabling people to deliberate about the decision information in accord with their personal values, which is associated with a better decision experience.


The Age of Blood Evaluation (ABLE) randomized controlled trial: study design.
Red blood cells (RBCs) are transfused to treat anemia and to maintain oxygen delivery to vital organs during critical illness. Laboratory and observational studies have raised the possibility that prolonged RBC storage may adversely affect clinical outcomes. Compared with RBCs stored less than 1 week, there are no clinical data demonstrating that RBCs stored longer remain as effective at carrying or releasing oxygen, and observational studies have risen to possibility that prolonged RBC storage might result in harm to vulnerable patients requiring blood transfusions. The "Age of Blood Evaluation" (ABLE) study (ISRCTN44878718) is a double-blind, multicenter, parallel randomized controlled clinical trial. It will test the hypothesis that the transfusion of prestorage leukoreduced RBCs stored for 7 days or less (fresh arm) as compared with standard-issue RBCs stored, on average, 15 to 20 days (control arm) will lead to lower 90-day all-cause mortality and reduced morbidity in critically ill adults. We include adults in intensive care units (ICUs) who (1) have had a request for a first RBC unit transfusion during the first 7 days of ICU admission and (2) have an anticipated requirement for ongoing invasive and noninvasive mechanical ventilation exceeding 48 hours. Enrolled patients are randomized at the time of transfusion to receive either standard-issue RBC units or RBCs stored 7 days or less issued by the local hospital transfusion service. The primary outcome is 90-day all-cause mortality. Secondary outcomes include ICU and hospital mortality, organ failure, and serious nosocomial infections. With 2510 patients, we will be able to detect a 5% absolute risk reduction (from 25% to 20%). The ABLE study is currently enrolling patients in 23 university-affiliated and community-hospital ICUs across Canada; sites in France and United Kingdom are expected to start recruitment in 2011. Regardless of the results, ABLE study will have significant implications on the duration of RBC storage. A negative trial will reassure clinicians and blood bankers regarding the effectiveness and safety of standard-issue RBCs. A positive trial will have significant implications with respect to inventory management of RBCs given to critically ill adults with a high risk of mortality and will also prompt research to better understand the RBC storage lesion in the hopes of minimizing its clinical consequences through the development of better storage methods.


A Tale of Confusion From Overlapping Confidence Intervals.
In clinical research presentations, study results are commonly reported in the form of P values and confidence intervals as an estimate of association and treatment effect. The interpretation of confidence intervals that overlap can be confusing and difficult for the reader to draw clinically meaningful conclusions. In this brief report, we describe the basics of confidence intervals and present an example from a recently published randomized control trial to illustrate a common confusion that overlapping confidence intervals between the means of two independent groups may not necessarily reject the true significant difference of effect. It is recommended that investigators use the direct difference of means between groups for confidence interval estimation to reduce type II errors. Clinicians should interpret overlapping confidence intervals with caution and avoid the assumption that overlapping confidence intervals always implies a lack of difference of treatment effect to decide application of treatment.


Pulmonary vascular pressures of strenuously exercising thoroughbreds after administration of phenylbutazone.
To determine the effects of phenylbutazone administration on heart rate and right atrial and pulmonary vascular pressures in Thoroughbreds during rest and during exercise performed at maximal heart rate. 7 healthy, exercise-conditioned Thoroughbreds. Horses were studied on 3 occasions: without medication [control], after i.v. administration of phenylbutazone (4.4 mg/kg of body weight) at 12-hour intervals for 2 days and a final dose given 1 hour before exercise, and after i.v. administration of phenylbutazone for 2 days in the same manner, but with the final dose given 24 hours before exercise. Horses were studied at rest and during exercise performed at maximal heart rate on a treadmill. Right atrial and pulmonary vascular pressures were measured with catheter-tip manometers referenced at the point of the shoulder. We did not detect significant differences in heart rate or right atrial and pulmonary vascular pressures among values recorded when horses were not given medication and values recorded when phenylbutazone was administered by either regimen. Exercise-induced pulmonary hemorrhage occurred in 6 of the 7 horses regardless of whether phenylbutazone was administered or the dosage regimen used. In these Thoroughbreds, phenylbutazone treatment did not modify heart rate or right atrial and pulmonary vascular pressures at rest or during exercise capable of eliciting exercise-induced pulmonary hemorrhage. Thus, because phenylbutazone is a potent inhibitor of cyclooxygenase, prostaglandins probably do not play a role in mediating exercise-induced pulmonary hypertension in horses. Phenylbutazone administration did not modify the pulmonary capillary hypertension in the strenuously exercising Thoroughbreds, and therefore, is unlikely to alter the prevalence or severity of exercise-induced pulmonary hemorrhage in Thoroughbred race-horses.


An SEM evaluation of conditioned and bonded enamel following carbamide peroxide bleaching and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) treatment.
The study aimed to evaluate, (a) the surface morphology of acid etched/conditioned enamel following carbamide peroxide bleaching with/without casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) (Tooth Mousse (MI Paste); GC Corp., Tokyo, Japan) treatment; and b) the nature of the bonded resin-enamel interfaces formed with a self-etching primer adhesive. Twenty-four human adult molars were each sectioned into four, the specimens divided and treated according to four experimental groups: 1, no treatment; 2, 16% carbamide peroxide bleaching; 3, CPP-ACP paste; 4, bleaching and CPP-ACP paste. A self-etching primer adhesive (Clearfil SE Bond, CSE) was used. The specimens were further divided into four subgroups for etching/conditioning: A, CSE Primer only; B, 30-40% phosphoric acid and CSE primer; C, 15% EDTA and CSE primer; D, 20% polyacrylic acid and CSE primer. Bonded specimens were also prepared. The morphology of the etched/conditioned enamel surfaces and polished resin-enamel interfaces of the bonded specimens were observed by field-emission scanning electron microscopy (FE-SEM). Treatment with CPP-ACP did not inhibit phosphoric acid etching of enamel. Poorly defined enamel etch patterns were observed with the other conditioners in all the groups. The morphology of the bonded resin-enamel interfaces observed on FE-SEM for each acid etching/conditioning subgroup was similar in all the experimental groups, except after bleaching. Resin infiltration into enamel was observed with prior phosphoric acid etching and polyacrylic acid conditioning and was unaffected by experimental group treatments. The use of a CPP-ACP paste with or without prior bleaching did not inhibit enamel etching. Enamel etching/conditioning may help improve bonding efficiency of the self-etching primer adhesive after CPP-ACP treatment.


[NMR study of the state of water in the human lens during cataract development].
Water proton spin-spin relaxation times (T2) and the content of bound, "non-freezable" at -9 degrees C water in both normal human lenses and human lenses of different stages of cataract progression (cataracta incipiens, nondum matura, mature hypermatura) were measured by NMR spin echoes method. By the stage of cataracta nondum matura, increase of bound water content and simultaneous, almost half decrease of the relaxation time (T2), were observed. However, on the following stages of cataract evaluation (almost mature, mature cataracts) a gradual decrease of bound water content is noted, but only for the mature cataract stage the water content significantly differs from that of the normal one. On the stage of hypermature cataract the presence of two unexchanged with each other fractions of water is found. The obtained data are explained by lens protein reconstructions during the cataract progression.


Effective Subcritical Butane Extraction of Bifenthrin Residue in Black Tea.
As a natural and healthy beverage, tea is widely enjoyed; however, the pesticide residues in tea leaves affect the quality and food safety. To develop a highly selective and efficient method for the facile removal of pesticide residues, the subcritical butane extraction (SBE) technique was employed, and three variables involving temperature, time and extraction cycles were studied. The optimum SBE conditions were found to be as follows: extraction temperature 45 °C, extraction time 30 min, number of extraction cycles 1, and in such a condition that the extraction efficiency reached as high as 92%. Further, the catechins, theanine, caffeine and aroma components, which determine the quality of the tea, fluctuated after SBE treatment. Compared with the uncrushed leaves, pesticide residues can more easily be removed from crushed leaves, and the practical extraction efficiency was 97%. These results indicate that SBE is a useful method to efficiently remove the bifenthrin, and as appearance is not relevant in the production process, tea leaves should first be crushed and then extracted in order that residual pesticides are thoroughly removed.


Dominant and diet-responsive groups of bacteria within the human colonic microbiota.
The populations of dominant species within the human colonic microbiota can potentially be modified by dietary intake with consequences for health. Here we examined the influence of precisely controlled diets in 14 overweight men. Volunteers were provided successively with a control diet, diets high in resistant starch (RS) or non-starch polysaccharides (NSPs) and a reduced carbohydrate weight loss (WL) diet, over 10 weeks. Analysis of 16S rRNA sequences in stool samples of six volunteers detected 320 phylotypes (defined at >98% identity) of which 26, including 19 cultured species, each accounted for >1% of sequences. Although samples clustered more strongly by individual than by diet, time courses obtained by targeted qPCR revealed that 'blooms' in specific bacterial groups occurred rapidly after a dietary change. These were rapidly reversed by the subsequent diet. Relatives of Ruminococcus bromii (R-ruminococci) increased in most volunteers on the RS diet, accounting for a mean of 17% of total bacteria compared with 3.8% on the NSP diet, whereas the uncultured Oscillibacter group increased on the RS and WL diets. Relatives of Eubacterium rectale increased on RS (to mean 10.1%) but decreased, along with Collinsella aerofaciens, on WL. Inter-individual variation was marked, however, with >60% of RS remaining unfermented in two volunteers on the RS diet, compared to <4% in the other 12 volunteers; these two individuals also showed low numbers of R-ruminococci (<1%). Dietary non-digestible carbohydrate can produce marked changes in the gut microbiota, but these depend on the initial composition of an individual's gut microbiota.


Development and validation of a high-throughput whole cell assay to investigate Staphylococcus aureus adhesion to host ligands.
Staphylococcus aureus adhesion to the host's skin and mucosae enables asymptomatic colonization and the establishment of infection. This process is facilitated by cell wall-anchored adhesins that bind to host ligands. Therapeutics targeting this process could provide significant clinical benefits; however, the development of anti-adhesives requires an in-depth knowledge of adhesion-associated factors and an assay amenable to high-throughput applications. Here, we describe the development of a sensitive and robust whole cell assay to enable the large-scale profiling of S. aureus adhesion to host ligands. To validate the assay, and to gain insight into cellular factors contributing to adhesion, we profiled a sequence-defined S. aureus transposon mutant library, identifying mutants with attenuated adhesion to human-derived fibronectin, keratin, and fibrinogen. Our screening approach was validated by the identification of known adhesion-related proteins, such as the housekeeping sortase responsible for covalently linking adhesins to the cell wall. In addition, we also identified genetic loci that could represent undescribed anti-adhesive targets. To compare and contrast the genetic requirements of adhesion to each host ligand, we generated a S. aureus Genetic Adhesion Network, which identified a core gene set involved in adhesion to all three host ligands, and unique genetic signatures. In summary, this assay will enable high-throughput chemical screens to identify anti-adhesives and our findings provide insight into the target space of such an approach.


Meiofaunal deuterostomes from the basal Cambrian of Shaanxi (China).
Deuterostomes include the group we belong to (vertebrates) as well as an array of disparate forms that include echinoderms, hemichordates and more problematic groups such as vetulicolians and vetulocystids. The Cambrian fossil record is well-populated with representative examples, but possible intermediates are controversial and the nature of the original deuterostome remains idealized. Here we report millimetric fossils, Saccorhytus coronarius nov. gen., nov. sp., from an Orsten-like Lagerstätte from the earliest Cambrian period of South China, which stratigraphically are amongst the earliest of deuterostomes. The bag-like body bears a prominent mouth and associated folds, and behind them up to four conical openings on either side of the body as well as possible sensory structures. An anus may have been absent, and correspondingly the lateral openings probably served to expel water and waste material. This new form has similarities to both the vetulicolians and vetulocystids and collectively these findings suggest that a key step in deuterostome evolution was the development of lateral openings that subsequently were co-opted as pharyngeal gills. Depending on its exact phylogenetic position, the meiofaunal habit of Saccorhytus may help to explain the major gap between divergence times seen in the fossil record and estimates based on molecular clocks.


A study of motion perception in primary open angle glaucoma.
To study the features of the motion perception (MP) and explore the worth of the clinical application of MP test in primary open angle glaucoma (POAG). MPs were recorded from 41 patients (67 eyes) with POAG ang 56 normal subjects (112 eyes) by the MP software in PC compatible computer. Meanwhile, the MP ways and MP sites were analyzed and MP abnormal rate was compared to the clinical parameters and the visual function indices related to glaucoma. The results showed that the total abnormal rate of MP was 89.5% in POAG, and the abnormal rate was 81.6% in early stage of POAG, and MP abnormal degree had positive correlation with C/D, ocular tension and corrected loss variance (CLV), and negative correlation with visual acuity and mean sensitivity (MS), and no correlation with age and short-term fluctuation (SF) in the patients with POAG. These results suggest that the MP test provides a newly effective examination method in diagnosing the early POAG.


Familial tumoral calcinosis and testicular microlithiasis associated with a new mutation of GALNT3 in a white family.
Familial tumoral calcinosis (FTC) is a rare autosomal recessive disease characterised by the development of multiple calcified masses in periarticular soft tissues; GALNT3 gene mutations have recently been described in an African American and in a Druse Arab family with FTC. To report the clinical and histological features caused by a new GALNT3 mutation in a white family. Homozygosity for the nonsense mutation Lys463X was found in both affected siblings, who displayed a classic phenotype, the male also having testicular microlithiasis. He is the first subject described with testicular microlithiasis in FTC. The high testicular expression of GALNT3 suggests that the gene alteration could act locally by causing deposition of calcium, and the testis may be an underestimated site of calcification in FTC. Autoimmune diseases are present in several members of the family. Although immune disorders have been described in FTC, autoimmunity does not segregate with the GALNT3 mutation in this family.


Neurological consequences of vitamin B12 deficiency and its treatment.
In developed countries, the vitamin B12 deficiency usually occurs in children exclusively breast-fed, whose mothers are vegetarians, causing low stores of vitamin B12. Symptoms of vitamin B12 deficiency appear during the second trimester of life and include failure to thrive, lethargy, hypotonia, and arrest or regression of developmental skills. A megaloblastic anemia can be present. One half of the infants exhibit abnormal movements before the start of treatment with intramuscular cobalamin, which disappear 1 or 2 days after. More rarely, movement disorders appear a few days after treatment, whereas neurological symptoms are improving. These abnormal movements can last for 2 to 6 weeks. If not treated, vitamin B12 deficiency can cause lasting neurodisability. Therefore, efforts should be directed to preventing deficiency in pregnant and breast-feeding women on vegan diets and their infants by giving them vitamin B12 supplements. When preventive supplementation has failed, one should recognize and treat quickly an infant presenting with failure to thrive and delayed development.


A comparison of ileal digesta and excreta analysis for the determination of amino acid digestibility in food ingredients for poultry.
1. The apparent ileal and excreta digestibilities of amino acids in 15 samples representing 12 food ingredients were determined using 5-week-old male broiler chickens. The ingredients included 3 samples of cereals (wheat, maize and sorghum), 6 samples of plant protein meals (soyabean meal, cottonseed meal, canola meal and sunflower meal) and 6 samples of animal protein meals (meat meal, meat-and-bone meal, feather meal and fish meal). 2. The test ingredients were incorporated as the sole source of dietary protein in assay diets. Each diet was offered ad libitum to 3 pens (4 birds/pen) from d 35 to d 42 post-hatching. Total collection of excreta was carried out during the last 4 d. All birds were killed on d 42 and the contents of the lower half of the ileum were collected. Apparent ileal and excreta amino acid digestibilities were calculated using acid-insoluble ash as the indigestible marker. 3. The influence of site of measurement was found to vary among food ingredients, among samples within an ingredient and among different amino acids within an ingredient. Ileal amino acid digestibility values were similar in some ingredients, but significantly lower or higher in others than the corresponding excreta values. 4. Average ileal and excreta amino acid digestibilities in sorghum and maize were similar, but significant differences were observed for individual amino acids. In contrast, ileal amino acid digestibility values were higher than the corresponding excreta digestibility values in wheat. 5. The average ileal and excreta digestibilites of amino acids in the 3 soyabean meal samples were similar although small, but significant differences were noted for individual amino acids. Site of measurement had no effect on the digestibility of amino acids in canola meal. Digestibilities of valine, isoleucine, phenylalanine, histidine, glutamic acid, alanine and tyrosine in sunflower meal and those of valine, methionine, isoleucine, leucine, lysine, glutamic acid and alanine in cottonseed meal were lower by excreta analysis. 6. Digestibilities in animal protein meals, with the exception of blood meal and fish meal, were consistently higher by excreta analysis. Ileal-excreta differences in individual amino acid digestibilities were more evident in feather meal, meat meal and meat-and-bone meal. 7. Threonine and valine were the indispensable amino acids that were more frequently influenced by the site of measurement. Of the dispensable amino acids, aspartic acid, serine, glutamic acid and alanine were the most affected. 8. Differences determined between ileal and excreta digestibilities in the present study clearly demonstrate that amino acid metabolism by hindgut microflora in chickens may be substantial and that digestibilities measured in the terminal ileum are more accurate measures of amino acid availability than those measured in the excreta.


Previous diagnosis of Sjögren's Syndrome as rheumatoid arthritis or systemic lupus erythematosus.
The diagnosis of SS is often difficult and many patients are symptomatic for years with other diagnoses before confirmation of SS. Our aim was to determine whether overlapping clinical and serologic features with RA and SLE may in part drive the misdiagnoses. A total of 1175 sicca patients were evaluated in a multidisciplinary clinic and classified as having SS based on the American-European Consensus Group Criteria. They were interrogated for a past history of suspicion or diagnosis of RA, SLE or SSc. These diseases were confirmed or ruled out by applying the corresponding classification criteria if the patients responded affirmatively. Of these, 524 (44.6%) subjects reported previous diagnosis or suspicion of RA, SLE or SSc, which was confirmed in 130 (24.8%) but excluded in 394 (75.2%) subjects. Of those previously diagnosed with another illness, 183 (34.9%) met the criteria for primary SS. RF was present in 70/191 patients with previous diagnosis of RA compared with 445/845 without a prior RA diagnosis (P = 3.38E-05), while 128/146 with a diagnosis of SLE had positive ANA compared with 622/881 without the diagnosis (P = 8.77E-06). Age also influenced former diagnoses: people with suspected RA were older than those without the diagnosis (P = 5.89E-06), while patients with SLE suspicion were younger (P = 0.0003). Interestingly, the previous diagnoses did not significantly delay a final classification of SS. Among subjects classified as SS, the presence of a positive ANA or RF was associated with a previous, apparently erroneous diagnosis of SLE or RA, respectively.


Occupational posttraumatic stress disorder and workplace violence in workers' compensation claims.
Acts of violence are the fifth leading cause of nonfatal occupational injuries in the United States. Experiencing a traumatic event at work can have serious mental health consequences, including the development of posttraumatic stress disorder (PTSD). This study aimed to quantify the prevalence of PTSD caused by workplace violence (WPV) in a statewide workers' compensation system and compare the outcomes and treatment of WPV cases versus those caused by other traumatic events. Using a retrospective cohort study design, workers who reported PTSD as the primary reason for a workers' compensation claim and had no coexisting physical injuries were found in California during 2009-2018. A total of 3,772 PTSD cases were identified, 48.9% of which were attributed to WPV. Demographic risk factors associated with WPV PTSD included lower income, younger age, female gender, and employment in retail or finance, p < .001-p = .007. For individuals who returned to work, claims due to WPV resulted in longer medically approved time away from work than non-WPV causes (Mdn = 132.5 days vs. Mdn = 91 days, respectively), p < .001. Three of the top 10 most frequently prescribed medications were administered against evidence-based guidelines. This study found that many treatments prescribed to PTSD patients are based on insufficient evidence, and the provision of existing empirically supported treatments is needed, particularly in generalized populations. The findings support the need for additional recognition of the cause of workplace PTSD to facilitate appropriate referrals to WPV or PTSD specialists to support return-to-work efforts.


A divide and conquer deformable contour method with a model based searching algorithm.
A divide and conquer deformable contour method is presented with an initial inside closed contour being divided into arbitrary segments, and these segments are allowed to deform separately preserving the segments' connectivity. A maximum area threshold, A/sub max/, is used to stop these outward contour segments' marching. Clear and blur contour points are then identified to partition the whole contour into clear and blur segments. A bi-directional searching method is then recursively applied to each blur segment including a search for contour-within-contour segment to reach a final close contour. Further improvements are provided by a model based searching algorithm. It is a two-step process with step 1 being a linked contour model matching operation where landmarks are extracted, and step 2 being a posteriori probability model matching and correction operation where large error segments are fine tuned to obtain the final results. The experiments include ultrasound images of pig heart, MRI brain images, MRI knee images having complex shapes with or without gaps, and inhomogeneous interior and contour region brightness distributions. These experiments have shown that the method has the capability of moving a contour into the neighboring region of the desired boundary by overcoming inhomogeneous interior, and by adapting each contour segment searching operation to different local difficulties, through a contour partition and repartition scheme in searching for a final solution.


In vivo efficacy of trovafloxacin against Bacteroides fragilis in mixed infection with either Escherichia coli or a vancomycin-resistant strain of Enterococcus faecium in an established-abscess murine model.
The pharmacodynamic and pharmacokinetic properties of trovafloxacin were studied in a standardized murine model of established subcutaneous abscesses. Daily dosing regimens of 37.5 to 300 mg/kg every 8 h (q8h) or every 24 h (q24h) were started 3 days after inoculation with mixtures containing either Bacteroides fragilis-Escherichia coli-autoclaved cecal contents (ACC) or B. fragilis-vancomycin-resistant Enterococcus faecium (VREF)-ACC. Treatment was continued for 3 or 5 days. The efficacy of treatment was determined by the decrease in abscess bacterial counts and abscess weights, as well as by the reduction in inflammation (biodistribution of (99m)Tc-HYNIC immunoglobulin G) compared to saline-treated controls. Trovafloxacin showed a significant dose-response effect on the bacterial counts, weight, and inflammation of B. fragilis-E. coli abscesses after 3 and/or 5 days of treatment. A maximum 3.4 and 3.1 log(10) reduction in CFU/abscess in the respective B. fragilis and E. coli bacterial counts was attained after 5 days of treatment with daily doses of 300 mg/kg. The peak serum concentration was more predictive for effect than the area under the concentration-time curve. The C(max) was the pharmacodynamic index most predictive for success, and the efficacy of the q24h regimens was significantly better than the q8h regimens. The antibiotic was ineffective against the VREF in mixed infection with B. fragilis, while the killing of the anaerobe in the same combination was significantly less than in the E. coli combination (P < 0.05). We conclude that this is a useful model for studying the activity of antimicrobials for the treatment of small (<1-cm), undrainable, mixed-infection abscesses. In addition, we have shown for the first time that a decrease in bacterial numbers also leads to a reduction in both abscess weight and inflammation.


Cephalometric Evaluation Based on Steiner's Analysis on Adults of Bihar.
To evaluate the mean cephalometric values for Steiner Analysis in bihar population. To evaluate the cephalometric features of a bihar population and to present_an organized, comprehensive cephalometric norms for Steiner cephalometric analysis.. To introduce mean values to assess skeletal, dental and soft tissue relationship using Steiner analysis for orthodontic diagnosis.. To compare standards that will be derived with the earlier established norms for other population. The study was conducted in the Department of orthodontics and dentofacial orthopedics,Buddha Institute of Dental Sciences And Hospital,Patna(Bihar). The sample was collected from the out patient department of the same college. The present analysis was made on lateral cephalograms of 60 subjects with well balanced and acceptable facial profile of Bihar population. The subjects were informed about the purpose of the study. Subjects with age group of 18-26 years with dento-alveolar class I malocclusion and acceptable facial profile were selected by trained orthodontists, who belonged to the department of orthodontics and dentofacial orthopedics,Buddha Institute of Dental Sciences And Hospital. There was increase in SNA angle, SNB angle, SND angle,in Bihar population than Caucasians. Bihar population has proclined and forwardly placed upper and lower incisors, which was indicative of bimaxillary protrusion as compared to Caucasians. Bihar population had protrusive upper and lower lips as compared to Caucasians. Bihar population males had tendency towards horizontal growth pattern of the mandible than females. Bihar females had more proclined lower incisors than males.


Self-concept, social support and hysterectomy.
This paper reports on part of a larger study financed by the DHSS and concerned with describing recovery from hysterectomy and evaluating an experimental counselling session aimed at helping women to plan and manage their own recovery. The effect of the operation on the woman's self-concept and the possible role of social support in the recovery process are the principal foci. Contrary to findings in much previous research, women in this study showed no evidence of adverse changes in self-concept, depression levels were reduced after operation, and the majority of women were glad to have had a hysterectomy. Low levels of support from partner, family and friends were associated with poorer outcome on some of the indicators used, and possible explanations for this are considered.


Developing post-stroke psychometric properties for an Adherence to rehabilitation regimen scale: A sequential exploratory mixed-method study.
Adherence to rehabilitation in patients post-stroke plays a significant role in the effectiveness of rehabilitation and patient recovery. This study aimed to design and determine the psychometric properties of a scale for measuring adherence to the rehabilitation regimen in patients post-stroke in the Iranian community. The present study used a sequential exploratory mixed method and was conducted in two phases (phase one qualitative and phase two quantitative). Participants in the first phase were patients post-stroke, caregivers, and rehabilitation team members (n=20). The second phase was conducted on patients post-stroke (n=198), and the psychometric steps, including face, content, and construct validity, as well as reliability, were assessed. The Adherence to Rehabilitation Regimen Scale (ARRS)was designed with 26 items and four factors of participation: 1) physical exercises, 2) following prescribed regimens, 3) performing the activities of daily living, and 4) psychological follow-up. The internal consistency was 0.96 by calculating Cronbach's alpha coefficient. The Interclass Correlation Coefficient was 0.99 with a confidence interval of 0.96-0.99. The scale measuring adherence to the rehabilitation regimen in patients post-stroke has optimal psychometric properties. Therefore, as the first specific scale to measure the degree of rehabilitation regimen adherence in patients post-stroke, this tool may be beneficial for other rehabilitation programs interested in managing and improving program adherence.


Fast inexact mapping using advanced tree exploration on backward search methods.
Short sequence mapping methods for Next Generation Sequencing consist on a combination of seeding techniques followed by local alignment based on dynamic programming approaches. Most seeding algorithms are based on backward search alignment, using the Burrows Wheeler Transform, the Ferragina and Manzini Index or Suffix Arrays. All these backward search algorithms have excellent performance, but their computational cost highly increases when allowing errors. In this paper, we discuss an inexact mapping algorithm based on pruning strategies for search tree exploration over genomic data. The proposed algorithm achieves a 13x speed-up over similar algorithms when allowing 6 base errors, including insertions, deletions and mismatches. This algorithm can deal with 400 bps reads with up to 9 errors in a high quality Illumina dataset. In this example, the algorithm works as a preprocessor that reduces by 55% the number of reads to be aligned. Depending on the aligner the overall execution time is reduced between 20-40%. Although not intended as a complete sequence mapping tool, the proposed algorithm could be used as a preprocessing step to modern sequence mappers. This step significantly reduces the number reads to be aligned, accelerating overall alignment time. Furthermore, this algorithm could be used for accelerating the seeding step of already available sequence mappers. In addition, an out-of-core index has been implemented for working with large genomes on systems without expensive memory configurations.


High-resolution structural analysis of chromatin at specific loci: Saccharomyces cerevisiae silent mating-type locus HMRa.
Genetic and biochemical evidence implicates chromatin structure in the silencing of the two quiescent mating-type loci near the telomeres of chromosome III in yeast. With high-resolution micrococcal nuclease mapping, we show that the HMRa locus has 12 precisely positioned nucleosomes spanning the distance between the E and I silencer elements. The nucleosomes are arranged in pairs with very short linkers; the pairs are separated from one another by longer linkers of approximately 20 bp. Both the basic amino-terminal region of histone H4 and the silent information regulator protein Sir3p are necessary for the organized repressive chromatin structure of the silent locus. Compared to HMRa, only small differences in the availability of the TATA box are present for the promoter in the cassette at the active MATa locus. Features of the chromatin structure of this silent locus compared to the previously studied HMLalpha locus suggest differences in the mechanisms of silencing and may relate to donor selection during mating-type interconversion.


Ca(2+)-modulated phosphorylation of a low-molecular-mass polypeptide in rat liver mitochondria: evidence that it is identical with subunit c of F(0)F(1)-ATPase.
A 3.5-kDa polypeptide associated with the inner membrane of rat liver was found to be phosphorylated by [gamma-(32)P]ATP, presumably via a cAMP-dependent kinase. The phosphorylation was modulated by [Ca(2+)] in the physiological range, with a minimum at 1 microM and rising fourfold toward lower (10 nM) and higher (10 microM) concentrations. Further characterization of the 3.5-kDa component showed that the polypeptide has the same electrophoretic mobility as subunit c of F(0)F(1)-ATPase and that it selectively binds to antibodies against subunit c.


Predicting time on prolonged benefits for injured workers with acute back pain.
Some workers with work-related compensated back pain (BP) experience a troubling course of disability. Factors associated with delayed recovery among workers with work-related compensated BP were explored. This is a cohort study of workers with compensated BP in 2005 in Ontario, Canada. Follow up was 2 years. Data was collected from employers, employees and health-care providers by the Workplace Safety and Insurance Board (WSIB). Exclusion criteria were: (1) no-lost-time claims, (2) >30 days between injury and claim filing, (3) <4 weeks benefits duration, and (4) age >65 years. Using proportional hazard models, we examined the prognostic value of information collected in the first 4 weeks after injury. Outcome measures were time on benefits during the first episode and time until recurrence after the first episode. Of 6,657 workers, 1,442 were still on full benefits after 4 weeks. Our final model containing age, physical demands, opioid prescription, union membership, availability of a return-to-work program, employer doubt about work-relatedness of injury, worker's recovery expectations, participation in a rehabilitation program and communication of functional ability was able to identify prolonged claims to a fair degree [area under the curve (AUC) = .79, 95% confidence interval (CI) .74-.84]. A model containing age, sex, physical demands, opioid prescription and communication of functional ability was less successful at predicting time until recurrence (AUC = .61, 95% CI .57, .65). Factors contained in information currently collected by the WSIB during the first 4 weeks on benefits can predict prolonged claims, but not recurrent claims.


Immobilization mechanisms of heavy metals by utilizing natural cow bone waste for municipal solid waste incineration fly ash treatment.
Municipal solid waste incinerator (MSWI) fly ash poses intricate compositional challenges and potential environmental hazards. Effective management of such hazardous waste is imperative to mitigate the release of toxic compounds into the environment. Solidification/stabilization (S/S) processes have emerged as a viable strategy to transform MSWI fly ash from incineration waste into a safer and more environmentally benign material. This study aims to comprehensively investigate the potential of utilizing cow bone waste to stabilize heavy metals, focusing on Pb, within municipal solid waste incineration fly ash. Experimental investigations encompassed cow bone-to-fly ash weight ratios ranging from 0.0 (control group) to 7:3, a settling time of 2 h, and a liquid-to-solid (L/S) ratio of 1.0 mL/g. Cow bone waste exhibited pronounced efficacy, particularly within the short settling time, yielding a remarkable Pb removal efficiency of up to 99% at a cow bone waste dose of merely 2% and an L/S ratio of 1.0 mL/g. Concurrently, other heavy metals such as Cd, Cu, and Zn were effectively stabilized with a cow bone waste dose of 1.5% during the same 2-h settling period. The results underscore the pivotal roles of ash/bone ratio and settling time in augmenting Pb stabilization in MSWI fly ash. The application of cow bone waste is anticipated to offer a cost-effective and environmentally sound approach, aligning with sustainable waste management principles.


Recent achievements and research initiated in the Swedish plastics and rubber industry.
The improvement in exposure conditions in the Swedish vinyl chloride producing industry is reported. The article comments on the technology and control methods by which the vinyl chloride concentration has been lowered to less than 1 ppm vinyl chloride. Two epidemiological retrospective cohort studies are presently under way on workers in PVC-utilizing industries and in the rubber industry.


Proteolytic cleavage confers nitric oxide synthase inducing activity upon prolactin.
Prolactin (PRL), originally associated with milk secretion, is now known to possess a wide variety of biological actions and diverse sites of production beyond the pituitary. Proteolytic cleavage is a common post-translational modification that can either activate precursor proteins or confer upon the peptide fragment unique biological actions not exerted by the parent molecule. Recent studies have demonstrated that the 16-kDa N-terminal proteolytic cleavage product of PRL (16K-PRL) acts as a potent inhibitor of angiogenesis. Despite previous demonstrations of 16K-PRL production in vivo, biological functions beyond its antiangiogenic actions remain unknown. Here we show that 16K-PRL, but not full-length PRL, acts to promote the expression of the inducible isoform of nitric oxide synthase (iNOS) and nitric oxide (*NO) production by pulmonary fibroblasts and alveolar type II cells with potency comparable with the proinflammatory cytokines interleukin-1beta, interferon gamma, and tumor necrosis factor alpha. The differential effect of 16K-PRL versus PRL occurs through a receptor distinct from known PRL receptors. Additionally, pulmonary fibroblasts express the PRL gene and endogenously produce 16K-PRL, suggesting that this pathway may serve both autocrine and paracrine roles in the regulation of *NO production. These results reveal that proteolytic cleavage of PRL confers upon this classical hormone potent iNOS inducing activity, suggesting its role in inflammatory/immune processes.


Incentivizing preventive services in primary care: perspectives on Local Enhanced Services.
General practitioners in the UK play a key role in prevention but provision of preventive services is variable. The 2004 General Medical Services contract allows Primary Care Trusts (PCTs) to address health needs through providing locally agreed payments for Local Enhanced Services (LESs). This study identifies how this contractual flexibility is used for preventive services and explores its perceived effectiveness. Semi-structured interviews were carried out (2008-09) in 10 purposively selected case study sites in England. Details of LESs for these sites were collected (2009) through Freedom of Information requests or local contacts. A national on-line survey of PCTs (2009) provided a national context for case study findings. LESs were considered to be effective in incentivizing preventive activity. However, specifications and performance management were often weak, awareness of how to optimize incentives was low and, as optional services, LESs were perceived to be at risk in a financial downturn. Using LESs for preventive services highlights gaps in 'core' primary care responsibilities and in the national pay-for-performance framework. Current incentive arrangements are complex, could increase inequalities and provide only a partial, short-term solution to developing a proactive approach to prevention in primary care.


Hormone replacement therapy in pre-menopausal women undergoing bilateral salpingo-oophorectomy for benign disease: A review of practice.
Introduction The National Institute for Health and Care Excellence (NICE) guideline on the diagnosis and management of the menopause states that women who are likely to go through the menopause as a result of surgical treatment should be offered information about the menopause and the importance of starting hormonal replacement therapy before they have their treatment. Objectives To determine compliance with NICE guidelines at the Royal Derby Hospital. Study design We undertook a retrospective review of all pre-menopausal women undergoing bilateral salpingo-oophorectomy for benign pathology between 1 January 2016 and 30 June 2016. Results Thirty-six cases were reviewed. Women were aged between 32.5 and 49.8 years old (median 45.13, inter-quartile range 42.6-47.6). The commonest indications for bilateral salpingo-oophorectomy were dysfunctional uterine bleeding (36.1%), chronic pelvic pain (30.5%), complex cyst (13.9%), and pre-menstrual syndrome (13.9%). Fifteen women (41.7%) did not have hormonal replacement therapy discussed. Only two (5.6%) had hormonal replacement therapy discussed pre-operatively. The remaining 19 had hormonal replacement therapy discussed post-operatively, either on the ward prior to discharge (n = 3) or, more commonly, in clinic six to eight weeks later. Although hormonal replacement therapy was only contraindicated in one woman (3%), it was only prescribed to five (24%). Results were slightly better for women under 40 but still only 28.6% had hormonal replacement therapy discussed with them pre-operatively. Conclusion This audit has demonstrated that compliance with the NICE guidelines is poor. We suspect similar results might be found in other gynaecology departments nationally. A hospital guideline to aid clinicians and a patient information sheet to educate women has been devised. A re-audit is planned six months after ratification of these documents.


Genes affecting the expression of cytochrome c in yeast: genetic mapping and genetic interactions.
The four mutant genes, cyc2, cyc3, cyc8 and cyc9, that affect the levels of the two iso-cytochromes c in the yeast Saccharomyces cerevisiae have been characterized and mapped. Both cyc2 and cyc3 lower the amount of iso-1-cytochrome c and iso-2-cytochrome c; whereas, cyc8 and cyc9 increase the amount of iso-2-cytochrome c. The cyc2, cyc3, cyc8 and cyc9 genes are located, respectively, on chromosomes XV, I, II and III, and are, therefore, unlinked to each other and unlinked to CYC1, the structural gene of iso-1-cytochrome c and to CYC7, the structural gene of iso-2-cytochrome c. While some cyc3 mutants are completely or almost completely deficient in cyotchromes c, none of the cyc2 mutants contained less than 10% of parental level of cytochrome c even though over one-half of the mutants contain UAA or UAG nonsense mutations. Thus, it appears as if a complete block of the cyc2 gene product still allows the formation of a residual fraction of cytochrome c. The cyc2 and cyc3 mutant genes cause deficiencies even in the presence of CYC7, cyc8 and cyc9, which normally cause overproduction of iso-2-cytochrome c. We suggest that cyc2 and cyc3 may be involved with the regulation or maturation of the iso-cytochromes c. In addition to having high levels of iso-2-cytochromes c, the cyc8 and cyc9 mutants are associated with flocculent cells and other abnormal phenotypes. The cyc9 mutant was shown to be allelic with the tup1 mutant and to share its properties, which include the ability to utilize exogenous dTMP, a characteristic flocculent morphology, the lack of sporulation of homozygous diploids and low frequency of mating and abnormally shaped cells of alpha strains. The diverse abnormalities suggest that cyc8 and cyc9 are not simple regulatory mutants controlling iso-2-cytochrome c.


Structural properties of monoclonal antibody aggregates induced by freeze-thawing and thermal stress.
Aggregation of monoclonal antibodies can be induced by freeze-thawing and elevated temperature, typical stress factors during development, production and storage. Our aim was to characterize structural properties of aggregates formed after freeze-thawing and thermal stressing of humanized monoclonal IgG(1) antibody (IgG). Formulations with 1.0mg/ml IgG in 100mM phosphate pH 7.2 were subjected to freeze-thawing and heating and characterized by spectroscopic techniques (UV-absorption, CD, ATR-FTIR and fluorescence), light obscuration, dynamic light scattering, SDS-PAGE, AF4 with UV and MALLS detection, and HP-SEC with UV and online fluorescent dye detection. Thermal stress led to an increased formation of dimers and soluble oligomers (HP-SEC, AF4). Aggregates smaller than 30nm were measured (DLS), next to slightly elevated particle levels in the mum range (light obscuration). Aggregates created by heating were in part covalently linked (SDS-PAGE) and made up of conformationally perturbed monomers (CD, ATR-FTIR, extrinsic dye fluorescence). Aggregation after freeze-thawing was manifested primarily in particle formation in the mum range. These aggregates were noncovalently linked (SDS-PAGE) and composed of native-like monomers, as obvious from CD, ATR-FTIR and extrinsic dye fluorescence spectroscopy. In conclusion, the complementary methods used in this study revealed that heating and freeze-thawing induced aggregates differ significantly in their physico-chemical characteristics.


The MoXFo initiative - adherence: Exercise adherence, compliance and sustainability among people with multiple sclerosis: An overview and roadmap for research.
We know very little about exercise adherence, compliance and sustainability in multiple sclerosis (MS), yet adherence is seemingly important for yielding immediate and sustained health benefits. This paper is focused on exercise adherence, compliance and sustainability in the context of informing research and practice involving MS. This focus is critical for clarifying terminology for future research and providing a roadmap guiding clinical research and practice. Our objective was accomplished through a narrative summary of the literature by a panel of experts on exercise adherence from the Moving Exercise Research in Multiple Sclerosis Forward (MoXFo) initiative and a concluding summary of the state of the literature and future research directions. The panel of experts identified three overall themes (Background and Importance; Understanding and Promoting Exercise Adherence, Compliance and Sustainability and Challenges to Exercise Adherence, Compliance and Sustainability) that represented a categorization of nine subthemes. These overall themes and subthemes formed the basis of our recommendations regarding future research broadly involving exercise adherence in MS. Overall, there is limited evidence on rates and determinants of exercise adherence and compliance in MS, and little is known about techniques and interventions for immediate and long-term exercise behaviour change.


Efficient transmission of 'Candidatus phytoplasma prunorum' Is delayed by eight months due to a long latency in its host-alternating vector.
Understanding at which spatiotemporal scale a disease causes significant secondary spread has both theoretical and practical implications. We investigated this issue in the case of European stone fruit yellows (ESFY), a quarantine vector-borne phytoplasma disease of Prunus trees. Our work was focused on the processes underlying disease spread: the interplay between the life cycles of the pathogen ('Candidatus Phytoplasma prunorum') and of the vector (Cacopsylla pruni). We demonstrated experimentally that C. pruni has only one generation per year and we showed that, at least in southeastern France, C. pruni migrates between conifers in mountainous regions (where it overwinters) and Prunus spp. at lower altitude (where it breeds). In acquisition-inoculation experiments performed with C. pruni over its period of presence on Prunus spp., both immature and mature C. pruni were hardly infectious (0.6%) despite effective phytoplasma acquisition and multiplication. We demonstrated that most immature vectors born on infected plants reach their maximum phytoplasma load (10(7) genomes per insect) only after migrating to conifers and that, after a life-long retention of the phytoplasma, their transmission efficiency was very high (60%) at the end of winter (when they migrate back to their Prunus host). Thus, most transmissions occur only after an effective latency of 8 months, following vector migrations and overwintering on conifers in mountainous regions. From this transmission cycle, we can infer that local secondary spread of ESFY in apricot orchards is marginal, and recommend that disease management strategies take more into account the processes occurring at a regional scale, including the role of wild Prunus spp. in ESFY epidemics.


Production, Characterization, and Function of Pseudoislets from Perinatal Canine Pancreas.
We evaluated the cell composition and function of canine pancreatic pseudoislets (PIs) produced from 42- to 55-day-old fetuses, 1- to 21-day-old pups, and an adult dog pancreas. After mild collagenase treatment, partially digested tissues were cultured for 2-3 weeks. PI production started on culture day 3, was marked for 6 to 9 days, and then stopped. PI production was greatest with the neonatal specimens, reaching about 12 million aggregates per litter (55-day-old fetus) or per pancreas (1-day-old pup). Cell composition at all stages was similar to that in adult pancreatic islets, with predominant β cells, scant α cells and, most importantly, presence of δ cells. Among pancreatic markers assessed by quantitative real-time PCR (qRT-PCR) mRNA assay, insulin showed the highest expression levels in PIs from newborn and adult pancreas, although these were more than 1000 times lower than in adult islets. Pdx1 mRNA expression was high in PIs from 55-day-old pancreases and was lower at later stages. Consistent with the qRT-PCR results, the insulin content was far lower than reported in adult dog pancreatic islets. However, insulin release by PIs from 1-day-old pups was demonstrated and was stimulated by a high-glucose medium. PIs were transplanted into euglycemic and diabetic SCID mice. In euglycemic animals, the transplant cell composition underwent maturation and transplants were still viable after 6 months. In diabetic mice, the PI transplants produced insulin and partially controlled the hyperglycemia. These data indicate that PIs can be produced ex vivo from canine fetal or postnatal pancreases. Although functional PIs can be obtained, the production yield is most likely insufficient to meet the requirements for diabetic dog transplantation without further innovation in cell culture amplification.


Comparison of total fluorine, extractable organic fluorine and perfluorinated compounds in the blood of wild and pefluorooctanoate (PFOA)-exposed rats: evidence for the presence of other organofluorine compounds.
The widespread occurrence and environmental persistence of perfluorinated compounds (PFCs) received worldwide attention recently. Exhaustive analysis of all fluorinated compounds in an environmental sample can be daunting because of the constraints in the availability of analytical standards and extraction methods. Combustion ion chromatographic technique for trace fluorine analysis was used to assess the concentrations of known PFCs (e.g., PFOS, PFOA) and total fluorine (TF) in the blood of wild rats collected from Japan. The technique was further validated using tissues from PFOA-exposed rats. Six PFCs (PFOS, PFOSA, PFUnDA, PFDA, PFNA, and PFOA) were detected in all of the wild rat blood samples. Concentrations of extractable organic fluorine (EOF) in fraction 1 (Fr1; MTBE extraction) of wild rats ranged 60.9-134 ng F mL(-1), while those in fraction 2 (Fr2; hexane) were below LOQ (32 ng F mL(-1)); TF concentrations in the blood of wild rats ranged from 59.9-192 ng F mL(-1). The contribution of known PFCs in EOF-Fr1 (MTBE) varied from 9% to 89% (56% on average), and known PFC concentrations in TF content were less than 25%. In contrast, TF concentrations in the blood of PFOA-exposed rats ranged from 46900 to 111,000 ng F mL(-1), with PFOA contributing over 90% of TF. A comparison of results from the samples analyzed in this study and the literature revealed three distinct groups with PFOA/known PFC and TF levels (i.e., wild rats and general population, occupationally exposed workers, and PFOA-exposed laboratory rats). The mass balance analysis of the different forms of fluorine in blood suggested the presence of other forms of organic fluorine in addition to known PFCs.


Cerebrospinal Fluid Treponema pallidum Particle Agglutination Assay for Neurosyphilis Diagnosis.
Limited data suggest that the cerebrospinal fluid Treponema pallidum particle agglutination assay (CSF-TPPA) is sensitive and a CSF Treponema pallidum hemagglutination assay (CSF-TPHA) titer of ≥1:640 is specific for neurosyphilis diagnosis. CSF-TPPA reactivity and titer were determined for a convenience sample of 191 CSF samples from individuals enrolled in a study of CSF abnormalities in syphilis (training data set). The sensitivity of a reactive test and the specificity for reactivity at serial higher CSF dilutions were determined. Subsequently, CSF-TPPA reactivity at a 1:640 dilution was determined for all available samples from study participants enrolled after the last training sample was collected (validation data set, n = 380). Neurosyphilis was defined as (i) a reactive CSF Venereal Disease Research Laboratory test (CSF-VDRL), (ii) detection of T. pallidum in CSF by reverse transcriptase PCR, or (iii) new vision loss or hearing loss. In the training data set, the diagnostic sensitivities of a reactive CSF fluorescent treponemal antibody absorption test (CSF-FTA-ABS) and a reactive CSF-TPPA did not differ significantly (67 to 98% versus 76 to 95%). The specificity of a CSF-TPPA titer of ≥1:640 was significantly higher than that of lower dilutions and was not significantly different from that of CSF-VDRL. In the validation data set, the diagnostic specificity of a CSF-TPPA titer of ≥1:640 was high and did not differ significantly from that of CSF-VDRL (93 to 94% versus 90 to 91%). Ten CSF samples with a nonreactive CSF-VDRL had a CSF-TPPA titer of ≥1:640. If a CSF-TPPA titer of ≥1:640 was used in addition to a reactive CSF-VDRL, the number of neurosyphilis diagnoses would have increased from 47 to 57 (21.3%). A CSF-TPPA titer cutoff of ≥1:640 may be useful in identifying patients with neurosyphilis when CSF-VDRL is nonreactive.


Intrinsic choroidal neurons in the human eye: projections, targets, and basic electrophysiological data.
The chemical coding of intrinsic choroidal neurons (ICNs) has features in common with extrinsic fibers (e.g., from the pterygopalatine ganglion) making it impossible to assess whether a neuronal nitric oxide synthase (nNOS)/vasoactive intestinal polypeptide (VIP)-immunoreactive nerve fiber is of intrinsic or extrinsic origin. Neurobiotin injections into single neurons allow the visualization of projections of these cells and the determination of the origin of target innervation. Thus, this technique was used in the present study to help characterize the organization of the ICN in the human eye. ICNs were visualized with the fluorescent vital dye 4-Di-2-ASP. Electrophysiological properties were determined by means of intracellular recordings. The impaled neurons were iontophoretically filled with neurobiotin. After fixation, immunohistochemistry for neuronal nitric oxide synthase (nNOS), alpha-smooth muscle actin, and calcitonin gene-related peptide (CGRP) was conducted. ICN processes were traced over distances of up to 2.612 micro m. They were found in the immediate vicinity of other nNOS-positive or -negative ICNs and were also found apposed to smooth muscle fibers (vascular and stromal nonvascular). CGRP-positive fibers forming boutons were observed closely associated with ICNs. Electrophysiological recording showed phasic firing without slow afterhyperpolarization, no spontaneous activity, an input resistance of 136 +/-73 MOmega, and a membrane time constant of 7 +/- 1 ms. Apart from the first functional characterization of ICNs, this study provided more precise evidence of reciprocal ICN-to-ICN contacts and innervation of both choroidal nonvascular and vascular smooth muscle. The presented technique offers promising perspectives to further investigate the function of ICNs in ocular homeostasis.


Effect of date seeds on oxidative damage and antioxidant status in vivo.
Date seeds have been shown to contain high amounts of antioxidants. However, in vivo studies on date seeds are lacking. Therefore the purpose of this study was to determine the effect of date seeds on oxidative damage and antioxidant status in vivo. Male Wistar rats were fed a basal diet containing 0, 70 or 140 g kg(-1) date seeds for 30 days. All three diets were isonitrogenous and isocaloric. Indication of oxidative damage was assessed in the liver and serum, and antioxidant status was assessed in the liver. Serum biochemical parameters, including indicators of tissue cellular damage and complete blood count with differential, were also determined. The results showed that date seeds significantly (P<0.05) reduced liver and serum malondialdehyde (a lipid peroxidative damage product) and serum lactate dehydrogenase and creatine kinase. Liver antioxidants (vitamin E, vitamin C, glutathione, superoxide dismutase, glutathione peroxidase and catalase), complete blood count with differential and other serum biochemical parameters assessed were not significantly altered by date seeds. The results obtained suggest a protective effect of date seeds against in vivo oxidative damage, possibly through the action of their bioactive antioxidants.


Sigmoidin F, a new prenylated flavonoid from Erythrina sigmoidea.
Sigmoidin F, a new prenylated flavanone, as well as abyssinone IV and 5,7,4'-trihydroxy-3'-methoxy-5'-(1"-prenyl) flavonone have been isolated from the stem and bark of Erythrina sigmoidea. The structure of 1 has been confirmed by a combination of 1H-nmr and other spectroscopic techniques.


Interventions to prevent HIV/AIDS among adolescents in less developed countries: are they effective?
The objective of this study was to summarize and critically assess the effects of interventions to prevent HIV/AIDS among adolescents in less developed countries. Reports of studies that evaluated interventions for preventing HIV/AIDS/STDs among persons aged 11-25 years were obtained from online computer databases, by searching conference proceedings and relevant journals, and by following up references cited in published reports. Studies were included if they investigated any educational, behavioral, psychosocial or other intervention that aimed to prevent or reduce HIV/AIDS/STD among persons aged 11-25 years in a less developed country. Only studies that included a control group, and which involved pre- and post-intervention assessments were included. Outcome measures included: (i) changes in safe sex practices (abstinence, condom use, limitation of sexual partners, avoidance of casual sex), (ii) knowledge about HIV/AIDS transmission and prevention methods, (iii) perception of HIV/AIDS/STD risks, (iv) self-efficacy with regard to condom negotiation and refusal of sex, (v) uptake of voluntary counseling and testing (VCT), and (vi) reduction in incidence of HIV/AIDS/STDs. Studies were assessed in terms of intervention format (e.g., education, role-play, video), duration, and setting (school or community). Reported improvements in outcome measures in intervention versus control groups were assessed. Sixteen studies met the inclusion criteria. Thirteen of these were conducted in Africa and three in Latin America. Twelve of the sixteen studies were school-based, and four were community-based. The interventions reviewed were not resoundingly successful in achieving their goals of increasing knowledge of HIV/AIDS, altering attitudes, improving negotiation and communication skills, or in influencing positive behavior evidenced through consistent condom use, abstinence, or reducing the number of partners. Considering the importance of HIV/AIDS prevention among adolescents, design of evaluation studies of programs in less developed countries need to be improved. The use of randomized controlled trials or other rigorous approaches for evaluating population-based behavioral interventions (e.g., Solomon Four design) is recommended.


[Protein composition and synthesis in islands of Langerhans isolated from normal rats and rats with alloxan diabetes].
Protein fractions with dissimilar electrophoretic mobility were identified in Langerhans, islands. In solitory islands, isolated from pancreas of rats with alloxan diabetes as compared with normal rats the protein composition varied distinctly: fractions of "constant" proteins and, specifically, the insulin fraction disappeared completely, total amount of identified fractions was decreased with relative alteration in their mobility. The total protein-synthesizing activity of Langerhans' islands from diabetic animals was 3-4-fold decreased as compared with that of control animals. An effect of "glucose repression" of protein synthesis was observed in isolated Langerhans' islands.


Perceived discrimination and psychotic experiences in the English general population.
Perceived discrimination has been linked to psychotic experiences (PEs). However, as yet, information is lacking on the relationship between different forms of discrimination and PEs. This study examined this association in the English general population. Nationally representative, cross-sectional data were analyzed from 7363 adults aged 16 and above that came from the Adult Psychiatric Morbidity Survey, 2007. Self-reported information was obtained on six forms of discrimination (ethnicity, sex, religious beliefs, age, physical health problems/disability, sexual orientation), while PEs were assessed with the Psychosis Screening Questionnaire (PSQ). Multivariable logistic regression analysis was used to assess associations. In a fully adjusted logistic regression analysis, any discrimination was significantly associated with PEs (odds ratio [OR]: 2.47, 95% confidence interval [CI]: 1.75-3.48). All individual forms of discrimination were significantly associated with PEs except sexual orientation. Multiple forms of discrimination were associated with higher odds for PEs in a monotonic fashion with those experiencing ≥ 3 forms of discrimination having over 5 times higher odds for any PE. In addition, experiencing any discrimination was associated with significantly increased odds for all individual forms of PE with ORs ranging from 2.16 (95%CI: 1.40-3.35) for strange experience to 3.36 (95%CI: 1.47-7.76) for auditory hallucination. Different forms of discrimination are associated with PEs in the general population. As discrimination is common at the societal level, this highlights the importance of public policy and evidence-based interventions to reduce discrimination and improve population mental health.


Lebetin 2, a Snake Venom-Derived B-Type Natriuretic Peptide, Provides Immediate and Prolonged Protection against Myocardial Ischemia-Reperfusion Injury via Modulation of Post-Ischemic Inflammatory Response.
Myocardial infarction (MI) followed by left ventricular (LV) remodeling is the most frequent cause of heart failure. Lebetin 2 (L2), a snake venom-derived natriuretic peptide, exerts cardioprotection during acute myocardial ischemia-reperfusion (IR) ex vivo. However, its effects on delayed consequences of IR injury, including post-MI inflammation and fibrosis have not been defined. Here, we determined whether a single L2 injection exerts cardioprotection in IR murine models in vivo, and whether inflammatory response to ischemic injury plays a role in L2-induced effects. We quantified infarct size (IS), fibrosis, inflammation, and both endothelial cell and cardiomyocyte densities in injured myocardium and compared these values with those induced by B-type natriuretic peptide (BNP). Both L2 and BNP reduced IS, fibrosis, and inflammatory response after IR, as evidenced by decreased leukocyte and proinflammatory M1 macrophage infiltrations in the infarcted area compared to untreated animals. However, only L2 increased anti-inflammatory M2-like macrophages. L2 also induced a higher density of endothelial cells and cardiomyocytes. Our data show that L2 has strong, acute, prolonged cardioprotective effects in post-MI that are mediated, at least in part, by the modulation of the post-ischemic inflammatory response and especially, by the enhancement of M2-like macrophages, thus reducing IR-induced necrotic and fibrotic effects.


Long-Term Association of Venous Thromboembolism With Frailty, Physical Functioning, and Quality of Life: The Atherosclerosis Risk in Communities Study.
Background Relatively little is known about the long-term consequences of venous thromboembolism (VTE) on physical functioning. We compared long-term frailty status, physical function, and quality of life among survivors of VTE with survivors of coronary heart disease (CHD) and stroke, and with those without these diseases. Methods and Results Cases of VTE, CHD, and stroke were continuously identified since ARIC (Atherosclerosis Risk in Communities Study) recruitment during 1987 to 1989. Functional measures were objectively captured at ARIC clinic visits 5 (2011-2013) and 6 (2016-2017); quality of life was self-reported. The 6161 participants at visit 5 were, on average, 75.7 (range, 66-90) years of age. By visit 5, 3.2% had had a VTE, 6.9% CHD, and 3.4% stroke. Compared with those without any of these conditions, VTE survivors were more likely to be frail (odds ratio [OR], 3.11; 95% CI, 1.80-5.36) and have low (<10) versus good scores on the Short Physical Performance Battery (OR, 3.59; 95% CI, 2.36-5.47). They also had slower gait speed, less endurance, and lower physical quality of life. VTE survivors were similar to coronary heart disease and stroke survivors on categorical frailty and outcomes on Short Physical Performance Battery assessment. When score on the Short Physical Performance Battery instrument was modeled continuously, VTE survivors performed better than stroke survivors but worse than CHD survivors. Conclusions VTE survivors had triple the odds of frailty and poorer physical function than those without the vascular diseases considered. Their function was somewhat worse than that of CHD survivors, but better than stroke survivors. These findings suggest that VTE patients may benefit from additional efforts to improve postevent physical functioning.


Evolution. Terrestrial life--fungal from the start?
Fungi have been implicated in the early colonization of land by plants. As Blackwell explains in her Perspective, Redecker et al.'s discovery of a fossil fungus provides strong support for this hypothesis. The important role of symbiotic associations of fungi with plants and animals is increasingly being recognized, as is the fact that some of these associations date back to the origin of terrestrial life on Earth.


Tumor marker CEA in monitoring of response to tegafur-uracil and folinic acid in patients with metastatic colorectal cancer.
The assessment of response to chemotherapy of solid tumors is generally made by measurement of tumors visualized by imaging, commonly computed tomography scanning. However, response assessment based on imaging is not always feasible because patients often have disease not measurable by imaging study, such as diffuse peritoneal dissemination. Furthermore, response assessment by imaging is expensive and time consuming. This study was carried out in an effort to evaluate the correlation between serial change on imaging and on CEA (carcinoembryonic antigen) levels for assessing chemotherapeutic response of patients with metastatic colorectal cancer. Between May 1998 and August 1999, a total of 40 patients with metastatic colorectal carcinoma were enrolled in this study. All the patients had to have measurable lesions. Oral tegafur-uracil 300 mg/m2/day and folinic acid 60 mg/day were administered concurrently for four weeks, repeated every five weeks, as the first-line treatment. Tumor marker CEA was examined before and during the whole course of treatment. Response based on CEA assessment was defined as a more-than 50% drop in serum CEA level for more than four weeks. The correlation between serial change on CEA and on imaging for assessing chemotherapeutic response was evaluated. Forty patients received a total of 318 courses of treatment and a response rate of 32.5% (95% confidence interval, 18.0% to 47.0%), including five complete responses and eight partial responses, was achieved by imaging studies. The pretreatment CEA levels were elevated beyond the normal cut-off value in 34 (85%) patients. The response rate evaluated by CEA assessment was 42.5% (17/40). Nine responders (22.5%) based on CEA had no remission on imaging. Agreement in assessment by imaging study and by CEA was observed in 20 patients (50%), including eight responders, five stable diseases, and seven progressive diseases. The sensitivity of falling CEA levels in the prediction of true responders on imaging was 62%. The sensitivity of elevated CEA levels for the prediction of progressive disease was 70%. Concerning the diagnostic accuracy, change in CEA levels in the prediction of true responders and progressive disease on imaging were 65% and 85%, respectively. On a follow-up of 24 months, patients with remarkable falling CEA levels survived significantly longer than non-responders (P = 0.0184, log-rank test). The measurement of CEA levels might be useful in monitoring chemotherapeutic response and in predicting the prognosis of patients with metastatic colorectal cancer. Serum CEA level may be used as a means of monitoring chemotherapeutic response when imaging study is unsuitable for assessing the response in clinical practice.


Measuring Hox-DNA binding by electrophoretic mobility shift analysis.
Understanding gene regulation by Hox transcription factors requires understanding the forces that underlie DNA binding by these proteins. Electrophoretic mobility shift analysis (EMSA) not only allows measurement of protein affinity and cooperativity but also permits visualization of differently migrating protein-DNA complexes, including complexes with different compositions or complexes with identical compositions yet assembled in different geometries. Furthermore, protein activity can be measured, allowing correction of binding constants for the percentage of protein that is properly folded and capable of binding DNA. Protocols for measuring protein activity and the equilibrium DNA-binding dissociation constant (K d) are provided. This versatile assay system can be adjusted based on specific needs to measure other parameters, including the kinetic association and dissociation constants (k a and k d) and the formation of heterologous protein-protein interactions.


Evidence of temperature-dependent sex determination in the European sea bass (Dicentrarchus labrax L.).
To test the hypothesis that sex determination in the European sea bass (Dicentrarchus labrax L.) Can be affected by the incubating temperature during the very early developmental stages, eggs from the same batch of spontaneously spawned broodstock were divided at the stage of half-epiboly into three groups according to rearing temperature: G13 = 13 degrees C, G15 = 15 degrees C, and G20 = 20 degrees C. Temperature treatment lasted until the middle of metamorphosis (17-18 mm total length, [TL]), and, with the exclusion of water temperature, all biotic and abiotic conditions were identical for the three experimental groups. The on-growing phase was performed under ambient photoperiod and temperature conditions for all groups. Sex proportions were determined by histological examination of the gonads of fish at 308, 467, and 568 days posthatch (DPH). At 308 DPH (TL: 135-201 mm), 100% of the specimens had differentiated into males and females. A significantly higher (P < 0.01) proportion of females was found in groups G13 (72-74%) and G15 (67-73%) than in group G20 (24-28%). At the final sampling there was no statistically significant difference in body weight between the experimental groups. However, in all groups, female fish were larger than males (P < 0.001). Results provide for the first time clear evidence that temperature during the very early developmental stages is the crucial factor affecting the process of sex differentiation of the sea bass, with low rearing temperatures (13 or 15 degrees C) resulting in sex proportions consistently skewed in favor of females.


Interactive gene expression pattern in prostate cancer cells exposed to phenolic antioxidants.
Dietary phenolic compounds are known to elicite vital cellular responses such as cell cycle arrest, apoptosis and differentiation by activating a cascade of molecular events. As there is an increasing interest to improve the efficacy of these compounds for use as potential chemopreventive agents, we wanted to understand the impact of phenolic compounds on target genes in prostate cancer. In this study we used human cDNA microarrays with 2400 clones consisting of 17 prosite motifs to characterize alterations in gene expression pattern in response to the phenolic antioxidants ellagic acid (EA) and resveratrol (RE). Over a 48-hr exposure of androgen - sensitive LNCaP cells to EA and RE, a total of 593 and 555 genes respectively, showed more than a two fold difference in expression. A distinct set of genes in both EA-and RE-treated cells may represent the signature profile of phenolic antioxidant-induced gene expression in LNCaP cells. Although extensive similarity was found between effects of EA - and RE - responsive genes in prostate cancer cells, out of 246 genes with overlapping responses, 25 genes showed an opposite effect. Quantitative RT-PCR was used to verify and validate the differential expression of selected genes identified from cDNA microarrays. In-depth analysis of the data from this study provided insight into the alterations in the p53 - responsive genes, p300, Apaf-1, NF-kBp50 and p65 and PPAR families of genes, suggesting the activation of multiple signaling pathways that leads to growth inhibition of LNCaP cells. This is a first study to look for changes in a large number of human genes in response to dietary compounds.


Optimized EGFR Blockade Strategies in EGFR Addicted Gastroesophageal Adenocarcinomas.
Gastric and gastroesophageal adenocarcinomas represent the third leading cause of cancer mortality worldwide. Despite significant therapeutic improvement, the outcome of patients with advanced gastroesophageal adenocarcinoma is poor. Randomized clinical trials failed to show a significant survival benefit in molecularly unselected patients with advanced gastroesophageal adenocarcinoma treated with anti-EGFR agents. We performed analyses on four cohorts: IRCC (570 patients), Foundation Medicine, Inc. (9,397 patients), COG (214 patients), and the Fondazione IRCCS Istituto Nazionale dei Tumori (206 patients). Preclinical trials were conducted in patient-derived xenografts (PDX). The analysis of different gastroesophageal adenocarcinoma patient cohorts suggests that EGFR amplification drives aggressive behavior and poor prognosis. We also observed that EGFR inhibitors are active in patients with EGFR copy-number gain and that coamplification of other receptor tyrosine kinases or KRAS is associated with worse response. Preclinical trials performed on EGFR-amplified gastroesophageal adenocarcinoma PDX models revealed that the combination of an EGFR mAb and an EGFR tyrosine kinase inhibitor (TKI) was more effective than each monotherapy and resulted in a deeper and durable response. In a highly EGFR-amplified nonresponding PDX, where resistance to EGFR drugs was due to inactivation of the TSC2 tumor suppressor, cotreatment with the mTOR inhibitor everolimus restored sensitivity to EGFR inhibition. This study underscores EGFR as a potential therapeutic target in gastric cancer and identifies the combination of an EGFR TKI and a mAb as an effective therapeutic approach. Finally, it recognizes mTOR pathway activation as a novel mechanism of primary resistance that can be overcome by the combination of EGFR and mTOR inhibitors.See related commentary by Openshaw et al., p. 2964.


Metal-loaded zeolite remediation of soils contaminated with pandrug-resistant Acinetobacter baumannii.
Due to the development of resistance to antimicrobial agents, bacterium Acinetobacter baumannii is nowadays a leading cause of nosocomial outbreaks. Clinically relevant A. baumannii outside hospital settings including natural soils affected by human waste represents a public-health risk for humans and animals. The aim of this study was to investigate the potential of metal-loaded zeolites to eliminate viable A. baumannii from artificially contaminated natural soils. A. baumannii isolate was subjected to the activity of natural zeolitised tuff (NZ) and Cu-modified (CuNZ) or Ag-modified zeolite (AgNZ) in wet, slightly acidic terra rossa and slightly alkaline red palaeosol. A. baumannii survived in terra rossa and red palaeosol supplemented with 1 wt% of NZ for seven days and four months, respectively. The addition of 1 wt% of CuNZ to terra rossa and red palaeosol shortened the survival of A. baumannii to three and 14 days, respectively. The addition of 0.1 wt% of AgNZ to both soils resulted in complete removal of viable A. baumannii within 1 h of contact, while the total native heterotrophic bacterial counts remained high. Since AgNZ is prepared with a simple modification of cost-effective and environmentally friendly natural zeolite, it is a promising material for the remediation of soils contaminated with pandrug-resistant A. baumannii.


The role of transferrin saturation as a screening test for hereditary haemochromatosis in an Irish population seeking medical care.
Genetic studies have indicated a high prevalence of mutations for hereditary haemochromatosis (HH) in Irish populations. Given the high cost of genetic screening and the ethical implications thereof, we assessed the role of transferrin saturation (TS) as a primary screening test for HH in an Irish population seeking medical care. TS and ferritin were measured on 330 consecutive blood specimens received in the laboratory for routine screening. Patients with TS > 45% were genetically screened for the C282Y and H63D mutations. Twenty-six patients had TS values > 45%. Of these, specimens were available for genetic screening on 20 patients. Three previously undiagnosed patients were found to be C282Y homozygotes and one a compound heterozygote (C282Y/H63D). The prevalence of C282Y homozygotes was 0.93%, which is similar to the reported prevalence found in Irish populations by genetic screening. Given the number of positive findings in this study, we conclude that, in the absence of a national programme, TS could be used as part of a health-screening panel in the Irish setting. Patients expressing iron overload would be detected before organ damage occurred, leading to less severe clinical disease and better patient prognosis.


Tau and HMW microtubule-associated proteins have different microtubule binding sites in vivo.
We have previously demonstrated the immunofluorescent localization of the tau and HMW MAPS on microtubules in vivo. When cells were treated with the non-ioni detergent triton before fixation, however treated with the non-ionic detergent triton before fixation, however, the immunofluorescent staining of microtubules with antiserum to HMW protein disappears, while the staining with anti-tubulin of anti-tau serum remains unaffected. Because of these differences in sensitivity to triton, we conclude that tau protein and HMW protein have different binding sites on microtubules. Parallel ultrastructural studies indicate that this loss of staining correlates with a loss of filamentous projections from the outer wall of cytoplasmic microtubules. These results indicate that in vivo the HMW protein is most probably distributed along the outer surface of the microtubule as filamentous projections similar to those observed in vitro. The in vivo distribution of the HMW protein suggests that it may form part of the microtrabecular system and be involved in the interaction of microtubules with other cell organelles.


The OECI model: the experience of INT Milan with a focus on the integration of psycho-oncology support.
The procedure for Organisation of European Cancer Institutes (OECI) accreditation identified the formation of MDTs for the diagnosis and scheduling of primary treatment with integration of supportive care as a key strength at Istituto Nazionale Tumori (INT). The opportunities for improvement highlighted by the OECI peer review inspired a study on the evaluation of psychological distress, with a view to integrating this evaluation into global patient management and to defining standardized criteria for the provision of psychological services. This article describes the OECI accreditation experience at INT and the study conducted between January and May 2015 on the evaluation of patients' distress levels during cancer treatment, defining a score-based cutoff point that triggers the intervention of a psychologist. The Distress Thermometer was used as a tool for evaluating psychological distress, performed by nurses on admitting the patient. A total of 261 questionnaires were completed by patients admitted to the medical oncology and hematology departments, with an average distress value of 4.1, and 60% of patients experiencing clinically detectable emotional distress. Emotion-related problems had a significant association with a greater level of distress, while there were few reports of relationship issues as a cause of distress. As a result of the improvement initiative supported by the psychological distress evaluation study, we validated the screening questionnaire to be included at the initial patient evaluation stage with a cutoff point triggering the intervention by a psychologist at a score of ≥7.


Chemoprevention of lung tumorigenesis by intranasally administered diindolylmethane in A/J mice.
The main reasons for the failure of most chemopreventive agents during clinical trials are poor in vivo bioavailability and dose-limiting side effects. One potential approach to surmount these problems in lung cancer chemoprevention trials could be direct delivery of agents into the pulmonary tissue. In this study, we assessed the efficacy of intranasally delivered bio-response diindolylmethane (BRD) against 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung tumorigenesis in mice. Mice treated with NNK (two doses of 50mg/kg at an interval of a week, intraperitoneal) developed 16.3±2.9 lung tumors per mouse. Post-carcinogen administration of BRD, via intranasal instillation, for 24 weeks, twice a week, at a dose of 2mg per mouse (0.6mg pure diindolylmethane per mouse) reduced the lung tumor multiplicity to 4.6±2.2 tumors per mouse (72% reduction). Likewise, large tumors (>1mm) were almost completely abolished and multiplicities of tumors with a size of 0.5-1mm were reduced by 74%. Tumor volume was also reduced by 82%. Further studies using an in vitro model of lung tumorigenesis showed that BRD exhibited pronounced antiproliferative and apoptotic effects in premalignant and malignant bronchial cells but only minimal effects in parental immortalized cells through, at least in part, suppression of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. These results showed the potent lung tumor inhibitory activities of low doses of BRD given via intranasal instillation and, therefore, intranasal delivery of BRD holds a great promise for lung cancer chemoprevention in subjects at high risk to develop lung cancer.