Study skills course in medical education for postgraduate residents.
The learners have to take active parts in the teaching learning activities. To make them aware and to help them develop the skills required, the need of the study skills course in medical education early in the part of their training has been realized for the postgraduate residents. The important areas of the study skills course focusing in the requirement of clinical components of the postgraduate residents are 1) Interpersonal and communications skills, 2) Teaching, learning and presentation skills, 3) Language, reading and computer use, 4) Evidence based medicine and diagnosis and management, 5) Assessment principles and strategies, 6) Time management strategies to get the best out of the training, 7) Reflection, portfolio and self-directed lifelong learning, and 8) Follow-up presentation. The methodologies that could be used in such study skills course are interactive lectures, brainstorming, presentations by the trainees, demonstration to and by the trainees, small group discussion, group work and presentation, group and individual feedback, practice sessions, role play, short relevant video movies, video recording of the trainees and viewing with feedback. With their already tight training schedule and posting and other similar other mandatory courses required for the postgraduate residents, much time cannot be allocated for the study skills course in medical education alone. Similar study skills course in medical education may need to be arranged for the undergraduate medical students as well.


Physical therapy improves venous hemodynamics in cases of primary varicosity: results of a controlled study.
Physical factors are known to influence hemodynamics in the veins of the lower extremities. In a controlled, randomized study the authors investigated the effects of combined physical therapy on varicose veins. Over a twenty-four-week period a treatment group consisting of 12 persons exercised under the supervision of a therapist twice a week for sixty minutes. This included muscle and joint activation by means of externally applied compression and cold-temperature stimuli (ie, thermosteresis). They also exercised once a day without supervision for fifteen minutes. During the same period a control group of 12 persons underwent the same measurements but no treatment. In the treatment group venous capacity decreased by an average of 16% from 4.9 +/- 0.3 (sd) mL/100 mL tissue to 4.1 +/- 0.4 (P < 0.005, U-test) while in the control group it remained practically unchanged at 4.8 +/- 0.4 vs 5.0 +/- 0.3. Venous refilling time in the lower extremities also increased in the treatment group, half refilling time rising from 7.8 +/- 1.0 to 11.3 +/- 0.9 seconds (P < 0.001) and total refilling time from 17.0 +/- 1.4 to 25.7 +/- 2.1 seconds (P < 0.001); these parameters remained virtually unchanged in the control group, with half refilling time dropping slightly from 7.7 +/- 1.1 to 7.1 +/- 1.3 seconds and total refilling time from 18.3 +/- 1.7 to 16.3 +/- 1.9 seconds. Patient self-rating scores obtained by use of a standardized questionnaire administered at baseline and at the end of week 24 improved in the treatment group only. The combined physical therapy was thus shown to be of long-term therapeutic value, improving venous function and reducing patients' symptoms. These findings indicate that for the further development of this combined treatment regimen it would be useful to identify the individual factors contributing to its efficacy and evaluate them separately.


Externalizing and internalizing problems in relation to autonomic function: a population-based study in preadolescents.
To investigate whether externalizing and internalizing problems are related to lower and higher heart rate (HR), respectively, and to explore the relationship of these problems with respiratory sinus arrhythmia (RSA) and baroreflex sensitivity (BRS). Moreover, to study whether problems present at both preschool and preadolescent age show stronger associations with autonomic function than those that were not. In a population cohort of 10- to 13-year-old children (N = 931; 11.6 +/- 0.5 years; 47% boys), autonomic measurements in supine and standing position were performed at school. RSA and BRS were determined by spectral analysis. Current externalizing and internalizing problems were assessed by the Child Behavior Checklist and problems at age 4 to 5 retrospectively by the Preschool Behavior Questionnaire. At supine rest, current externalizing problems were associated with lower HR and higher RSA, but not with BRS and current internalizing problems with higher HR and lower RSA, but not with BRS. These results were specifically found in children with problems that were retrospectively reported to have been also present at preschool age. Standing-induced changes in autonomic parameters were unrelated to the behavioral dimensions. Externalizing and internalizing problems are associated with divergent autonomic patterns, suggesting autonomic underarousal and overarousal, respectively. Problems starting early in life may specifically account for this. This should be confirmed in prospective studies.


Comorbidities in familial Mediterranean fever: analysis of 2000 genetically confirmed patients.
FMF is the most common periodic fever syndrome, characterized by recurrent episodes of fever and serosal inflammation accompanied with high acute phase reactants. The analysis of possible comorbidities is important to understand the impact of these conditions on clinical care and whether they share a common aetiological pathway. In this study, we aimed to evaluate the comorbidities associated with FMF patients in a large genetically diagnosed cohort. We retrospectively evaluated the medical and genetic records of FMF patients who were followed up by rheumatologists in Hacettepe University for 15 years. The FMF patients who had homozygous or compound heterozygous mutations were included in the study. Comorbidities associated with FMF were divided into three groups: (i) comorbidities directly related to FMF, (ii) comorbidities due to increased innate inflammation, and (iii) comorbidities that were regarded as being incidental. A total of 2000 patients with a diagnosis of FMF were enrolled in the study. Among them 636 were children (31.8%) and M694V was the most common mutation in patients with associated inflammatory conditions. The frequency of AS, Iga Vasculitis (Henoch-Schönlein purpura), juvenile idiopathic arthritis, polyarteritis nodosa, multiple sclerosis and Behçet's disease were increased in patients with FMF when compared with those in the literature. This study represents the largest genetically confirmed cohort and compares the frequencies with existing national and international figures for each disease. The increased innate immune system inflammation seen in FMF may be considered as a susceptibility factor since it predisposes to certain inflammatory conditions.


Genomics and epigenomics: new promises of personalized medicine for cancer patients.
Recent years have brought about a marked extension of our understanding of the somatic basis of cancer. Parallel to the large-scale investigation of diverse tumor genomes the knowledge arose that cancer pathologies are most often not restricted to single genomic events. In contrast, a large number of different alterations in the genomes and epigenomes come together and promote the malignant transformation. The combination of mutations, structural variations and epigenetic alterations differs between each tumor, making individual diagnosis and treatment strategies necessary. This view is summarized in the new discipline of personalized medicine. To satisfy the ideas of this approach each tumor needs to be fully characterized and individual diagnostic and therapeutic strategies designed. Here, we will discuss the power of high-throughput sequencing technologies for genomic and epigenomic analyses. We will provide insight into the current status and how these technologies can be transferred to routine clinical usage.


The future nephrology workforce: will there be one?
Interest in nephrology as a career is declining and has been on the decline for nearly one decade. From 2002 to 2009, all internal medicine subspecialties except geriatric medicine increased the number of available fellowship positions. However, only two subspecialties attracted fewer United States medical graduates (USMGs) in 2009 than in 2002: geriatric medicine and nephrology. This drop occurred at a time when demand for nephrologists is increasing and when the specialty is having a harder time benefiting from the substantial contribution of international medical graduates (IMGs). Today's USMGs possess fundamentally different career and personal goals from their teachers and mentors. Medical students report receiving minimal exposure to nephrology in clinical rotations, and they perceive that the specialty is too complex, uninteresting, and lacks professional opportunity. Meanwhile, the demographics of kidney disease in the United States, as well as recent national health policy developments, indicate a growing need for nephrologists. Efforts to improve the educational continuum in nephrology and enhance mentorship are essential to restoring interest in nephrology for USMGs, maintaining its appeal among IMGs, and developing a workforce sufficient to meet future demand for renal care.


Does Needle Design Affect the Regenerative Potential of Bone Marrow Aspirate? An In Vitro Study.
While autologous bone is still the gold standard for treatment of bone defects, its availability is limited. Sufficient numbers of mesenchymal stroma cells (MSC) may be an alternative. Small volumes of bone marrow aspirate (BMA) were harvested with two different needle systems comparing the yield and regenerative potency of the MSCs. BMA (10 mL) was aspirated from the posterior iliac crest of 12 patients with degenerative spinal disc disease using both needle systems in each patient: the Jamshidi needle (JAM) and on the contralateral side the Marrow Cellution® Needle (AMC). Number of mononuclear cells (MNCs) and regeneration capacity (colony-forming unit/CFU) were determined. MSCs were characterized for surface markers and their differentiation into trilineages. There was no significant difference between the two harvesting needles regarding the quantity of MNCs in BMA: 5.2 ± 1.8 × 109 MNC/mL for AMC vs. 4.8 ± 2.5 × 109 MNC/mL for JAM, p = 0.182. The quantity of CFUs per ml BMA was similar for both groups: 3717 ± 5556 for AMC and 4305 ± 5507 for JAM (p = 0.695). The potency of MSCs expressed as colony-forming potential per 106 MNC resulted in 0.98 ± 1.51 for AMC and 1.00 ± 0.96 for JAM (p = 0.666). Regardless of the needle design, 10 mL bone marrow aspirate contains a sufficient number of about 40,000 MSCs that can be used to enhance bone healing.


Decrease in platelet reduced glutathione increases lipoxygenase activity and decreases vitamin E.
Unstimulated normal human blood platelets were treated with azodicarboxylic acid bis(dimethylamide) (diamide), a thiol-oxidizing agent. Oxygenated arachidonic acid (AA) metabolites, malondialdehyde (MDA), and tocopherols were then quantified by high-performance liquid chromatography (HPLC). Diamide treatment partially decreased the amount of reduced glutathione (GSH) content and induced a subsequent decrease in peroxidase activity. However, formation of 12-hydroxy-eicosatetraenoic acid (12-HETE), the end-product of lipoxygenation of AA, increased. Formation of MDA, a marker of overall lipid peroxidation, was also enhanced. Furthermore, platelet alpha-tocopherol, but not gamma-tocopherol, significantly decreased. These results indicate that enhanced "basal" lipoxygenase activity, as a marker of specific AA oxygenation, may be linked to decreased platelet antioxidant status.


Activity of the Human Rhinovirus 3C Protease Studied in Various Buffers, Additives and Detergents Solutions for Recombinant Protein Production.
Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications.


Transgene-mediated skeletal phenotypic variation in zebrafish.
When considering relationships between genotype and phenotype we frequently ignore the fact that the genome of a typical animal, notably including that of a fish and a human, harbours a huge amount of foreign DNA. Such DNA, in the form of transposable elements, can affect genome function in a major way, and transgene biology needs to be included in our understanding of the genome. Here we examine an unexpected phenotypic effect of the chromosomally integrated transgene fli1a-F-hsp70l:Gal4VP16 that serves as a model for transgene function generally. We examine larval fras1 mutant zebrafish (Danio rerio). Gal4VP16 is a potent transcriptional activator that is already well known for toxicity and mediating unusual transcriptional effects. In the presence of the transgene, phenotypes in the neural crest-derived craniofacial skeleton, notably fusions and shape changes associated with loss of function fras1 mutations, are made more severe, as we quantify by scoring phenotypic penetrance, the fraction of mutants expressing the trait. A very interesting feature is that the enhancements are highly specific for fras1 mutant phenotypes, occurring in the apparent absence of more widespread changes. Except for the features due to the fras1 mutation, the transgene-bearing larvae appear generally healthy and to be developing normally. The transgene behaves as a genetic partial dominant: a single copy is sufficient for the enhancements, yet, for some traits, two copies may exert a stronger effect. We made new strains bearing independent insertions of the fli1a-F-hsp70l:Gal4VP16 transgene in new locations in the genome, and observed increased severities of the same phenotypes as observed for the original insertion. This finding suggests that sequences within the transgene, for example Gal4VP16, are responsible for the enhancements, rather than the effect on neighbouring host sequences (such as an insertional mutation). The specificity and biological action underlying the traits are subjects of considerable interest for further investigation, as we discuss. Our findings show that work with transgenes needs to be undertaken with caution and attention to detail.


Characterization of single stranded telomeric DNA-binding proteins in cultured soybean (Glycine max) cells.
We have identified and characterized a protein factor in soybean (Glycine max) nuclear extracts that binds to plant single stranded telomeric DNA repeats. A single DNA-protein complex was detected in gel retardation assays using synthetic telomeres and nuclear extracts. The protein forming this complex was designated soy-bean (Glycine max) single stranded telomeric DNA-binding protein (Gm-STBP). Gm-STBP binds to single stranded telomeric DNA containing more than two repeats. It does not bind to Tetrahymena, human or mutated plant telomere sequences, and its binding activity is not affected by RNase treatment. Gm-STBP activity gradually decreased after suspension cultures entered stationary phase. A slower migrating band was formed with extracts of earlier and later phases of soybean suspension cultures. Our findings suggest that binding of Gm-STBP to plant single stranded telomeric DNA may play a role in the proper functioning of telomeres during development.


Maternal parenting styles and mother-child relationship among adolescents with and without persistent attention-deficit/hyperactivity disorder.
We investigated mothering and mother-child interactions in adolescents with and without persistent attention-deficit/hyperactivity disorder (ADHD) in a sample of 190 adolescents with persistent DSM-IV ADHD, 147 without persistent ADHD, and 223 without ADHD. Both participants and their mothers received psychiatric interviews for diagnosis of ADHD and other mental disorders; and reported on the Parental Bonding Instrument about mother's parenting style, the Social Adjustment Inventory for Children and Adolescents for interactions with mothers and home behavioral problems. The mothers also reported on their ADHD and neurotic/depressive symptoms. Our results based on both informants showed that both ADHD groups obtained less affection/care and more overprotection and control from the mothers, and perceived less family support than those without ADHD. Child's inattention and comorbidity, and maternal depression were significantly correlated with decreased maternal affection/care and increased maternal controls; child's hyperactivity-impulsivity and maternal neurotic trait were significantly correlated with maternal overprotection; and child's inattention and comorbidity, and maternal neurotic/depressive symptoms were significantly correlated with impaired mother-child interactions and less family support. Our findings suggested that, regardless of persistence, childhood ADHD diagnosis, particularly inattention symptoms and comorbidity, combining with maternal neurotic/depressive symptoms was associated with impaired maternal process.


Organic matter lability modifies the vertical structure of methane-related microbial communities in lake sediments.
Eutrophication increases the input of labile, algae-derived, organic matter (OM) into lake sediments. This potentially increases methane (CH4) emissions from sediment to water through increased methane production rates and decreased methane oxidation efficiency in sediments. However, the effect of OM lability on the structure of methane oxidizing (methanotrophic) and methane producing (methanogenic) microbial communities in lake sediments is still understudied. We studied the vertical profiles of the sediment and porewater geochemistry and the microbial communities (16S rRNA gene amplicon sequencing) at five profundal stations of an oligo-mesotrophic, boreal lake (Lake Pääjärvi, Finland), varying in surface sediment OM sources (assessed via sediment C:N ratio). Porewater profiles of methane, dissolved inorganic carbon (DIC), acetate, iron, and sulfur suggested that sites with more autochthonous OM showed higher overall OM lability, which increased remineralization rates, leading to increased electron acceptor (EA) consumption and methane emissions from sediment to water. When OM lability increased, the abundance of anaerobic nitrite-reducing methanotrophs (Candidatus Methylomirabilis) relative to aerobic methanotrophs (Methylococcales) in the methane oxidation layer of sediment surface decreased, suggesting that Methylococcales were more competitive than Ca. Methylomirabilis under decreasing redox conditions and increasing methane availability due to their more diverse metabolism (fermentation and anaerobic respiration) and lower affinity for methane. Furthermore, when OM lability increased, the abundance of methanotrophic community in the sediment surface layer, especially Ca. Methylomirabilis, relative to the methanogenic community decreased. We conclude that increasing input of labile OM, subsequently affecting the redox zonation of sediments, significantly modifies the methane producing and consuming microbial community of lake sediments. IMPORTANCE Lakes are important natural emitters of the greenhouse gas methane (CH4). It has been shown that eutrophication, via increasing the input of labile organic matter (OM) into lake sediments and subsequently affecting the redox conditions, increases methane emissions from lake sediments through increased sediment methane production rates and decreased methane oxidation efficiency. However, the effect of organic matter lability on the structure of the methane-related microbial communities of lake sediments is not known. In this study, we show that, besides the activity, also the structure of lake sediment methane producing and consuming microbial community is significantly affected by changes in the sediment organic matter lability.


A SAW resonator filter using longitudinal and transverse modes.
A SAW resonator filter using not only transverse but also longitudinal modes is presented. The design principle permits to construct four-pole filters without cascading or parallel connection. The input and output transducers arranged side by side have equal construction characterized by unsymmetrical withdrawing weighting. Neighboring disturbing longitudinal modes are suppressed due to weighting determined by an iterative procedure. The successful application of the principle to a device is demonstrated by experimental measurements.


Lieno-adrenal shunt.
Portal hypertension with variceal bleed and a well functioning liver is an indication for shunt surgery. The commonly performed procedure is the lieno-renal shunt. When the left adrenal vein is long enough, and of healthy diameter a spleno-adrenal shunt is possible. Most of the reported cases are of distal spleno-adrenal type. A case of proximal spleno-adrenal patent shunt with a follow-up two years is presented along with review of literature.


Reactive surface organometallic complexes observed using dynamic nuclear polarization surface enhanced NMR spectroscopy.
Dynamic Nuclear Polarization Surface Enhanced NMR Spectroscopy (DNP SENS) is an emerging technique that allows access to high-sensitivity NMR spectra from surfaces. However, DNP SENS usually requires the use of radicals as an exogenous source of polarization, which has so far limited applications for organometallic surface species to those that do not react with the radicals. Here we show that reactive surface species can be studied if they are immobilized inside porous materials with suitably small windows, and if bulky nitroxide bi-radicals (here TEKPol) are used as the polarization source and which cannot enter the pores. The method is demonstrated by obtaining significant DNP enhancements from highly reactive complelxes [([triple bond, length as m-dash]Si-O-)W(Me)5] supported on MCM-41, and effects of pore size (6.0, 3.0 and 2.5 nm) on the performance are discussed.


The use of biomarkers in human pharmacology (Phase I) studies.
The development of a new medicine is a risky and costly undertaking that requires careful planning. This planning is largely applied to the operational aspects of the development and less so to the scientific objectives and methodology. The drugs that will be developed in the future will increasingly affect pathophysiological pathways that have been largely unexplored. Such drug prototypes cannot be immediately introduced in large clinical trials. The effects of the drug on normal physiology, pathophysiology, and eventually the desired clinical effects will need to be evaluated in a structured approach, based on the definition of drug development as providing answers to important questions by appropriate clinical studies. This review describes the selection process for biomarkers that are fit-for-purpose for the stage of drug development in which they are used. This structured and practical approach is widely applicable and particularly useful for the early stages of innovative drug development.


Development and testing of tools to detect ambulatory surgical adverse events.
Numerous health-care systems in the United States, including the Veterans Health Administration (VA), use the National Surgical Quality Improvement Program (NSQIP) to detect surgical adverse events (AEs). VASQIP sampling methodology excludes many routine ambulatory surgeries from review. Triggers, algorithms derived from clinical logic to flag cases where AEs have most likely occurred, could complement VASQIP by detecting a higher yield of ambulatory surgeries with a true surgical AE. We developed and tested a set of ambulatory surgical AE trigger algorithms using a sample of fiscal year 2008 ambulatory surgeries from the VA Boston Healthcare System. We used VA Boston VASQIP-assessed cases to refine triggers and VASQIP-excluded cases to test how many trigger-flagged surgeries had a nurse chart review-detected surgical AE. Chart review was performed using the VA electronic medical record. We calculated the ratio of cases with a true surgical AE over flagged cases (i.e., the positive predictive value [PPV]), and the 95% confidence interval for each trigger. Compared with the VASQIP rate (9 AEs, or 2.8%, of the 322 charts assessed), nurse chart review of the 198 trigger-flagged surgeries yielded more cases with at least 1 AE (47 surgeries with an AE, or 6.0%, of the 782 VASQIP-excluded ambulatory surgeries). Individual trigger PPVs ranged from 12.4% to 58.3%. In comparison with VASQIP, our set of triggers identified a higher rate of surgeries with AEs in fewer chart-reviewed cases. Because our results are based on a relatively small sample, further research is necessary to confirm these findings.


Exposure to a firefighting overhaul environment without respiratory protection increases immune dysregulation and lung disease risk.
Firefighting activities appear to increase the risk of acute and chronic lung disease, including malignancy. While self-contained breathing apparatuses (SCBA) mitigate exposures to inhalable asphyxiates and carcinogens, firefighters frequently remove SCBA during overhaul when the firegrounds appear clear of visible smoke. Using a mouse model of overhaul without airway protection, the impact of fireground environment exposure on lung gene expression was assessed to identify transcripts potentially critical to firefighter-related chronic pulmonary illnesses. Lung tissue was collected 2 hrs post-overhaul and evaluated via whole genome transcriptomics by RNA-seq. Although gas metering showed that the fireground overhaul levels of carbon monoxide (CO), carbon dioxide (CO2), hydrogen cyanine (HCN), hydrogen sulfide (H2S) and oxygen (O2) were within NIOSH ceiling recommendations, 3852 lung genes were differentially expressed when mice exposed to overhaul were compared to mice on the fireground but outside the overhaul environment. Importantly, overhaul exposure was associated with an up/down-regulation of 86 genes with a fold change of 1.5 or greater (p<0.5) including the immunomodulatory-linked genes S100a8 and Tnfsf9 (downregulation) and the cancer-linked genes, Capn11 and Rorc (upregulation). Taken together these findings indicate that, without respiratory protection, exposure to the fireground overhaul environment is associated with transcriptional changes impacting proteins potentially related to inflammation-associated lung disease and cancer.


Variations in human HM74 (GPR109B) and HM74A (GPR109A) niacin receptors.
HM74 (GPR109B) and the highly homologous gene, HM74A (GPR109A) code for Gi-G protein-coupled orphan receptors that recently have been discovered to be involved in the metabolic effects of niacin. The B vitamin niacin is an important agent used in the treatment of dyslipidemias, but its use is limited by side effects. The novel role of the adjacent HM74 and HM74A genes in the metabolism of niacin may provide new targets for drug development. Human genetic variations in HM74 and HM74A have been reported but have not been studied in detail. These variations may play a role in the response to agents targeting receptors coded by these genes. Here we show that many of the nonsynonymous SNPs listed in public databases for HM74 and HM74A are artifacts resulting from extensive homology between these two genes. This may be representative of a neglected phenomenon in reporting sequences of highly homologous genes. We provide primer sequences that permit selective amplification of the complete coding regions of HM74 and HM74A. Using these primers, we show that subsequent sequencing of HM74 and HM74A reveals a novel and unique variation in the HM74A gene. Haplotype analysis suggests four SNPs can define the five major haplotypes that lie within a single haplotype block encompassing these two genes.


[Human leptospirosis. A short review concerning a caseload].
Leptospirosis, a zoonosis with worldwide distribution, may have a broad spectrum of clinical manifestations that can delay the diagnosis, especially in wet climates where other conditions may dissimulate the main aspects. The authors describe 42 cases of patients with leptospirosis admitted to the Infectious Diseases Unit of a general hospital since 1990. The goal was to analyse epidemiological features, clinical manifestations, laboratory findings, treatment and evolution. There were 23 males (54.7%) and 19 females (45.2%), with ages ranging from 17 to 82 years. Most cases occurred in occupational settings, especially in those in contact with animals or stagnant water. All the cases had serological confirmation (by MAT or ELISA methods). Anicteric forms represented 42.9% of the total, which implies that in many cases a diagnosis is not immediately evident. The main symptoms, other than fever (97.6%) and jaundice (57.1%), were myalgia (71.4%), headache (42.8%) and nausea (33.3%). Renal involvement was common (52%), but the most severe forms occurred in 3 patients who required hemodialysis; followed by D.I.C. in 9.5%; meningitis in 4.7% and pulmonary involvement in 2.3% of the cases. The serovars most commonly identified were L. icterohaemorraghiae (28.5%), L. australis (14.3%), L. grippothyphosa (11.9%) and L. canicola (9.5%). The patients were treated with penicillin G (83.3%) or doxycyclin (9.5%). Two of them deceased (4.7%) and the remainder had a good evolution. The diagnosis of leptospirosis may be delayed if physicians fail to include this infection in the initial differential diagnosis. In view of this, the authors emphasize that when nonspecific clinical manifestations occur, a good epidemiological history is useful in proposing this diagnosis.


Expanded Use of Preloaded Branched and Fenestrated Endografts for Endovascular Repair of Complex Aortic Aneurysms.
The aim of this study was to report the expanded use of preloaded catheters and wires of fenestrations and directional branches to facilitate access to renal and mesenteric target arteries during endovascular repair of complex aortic aneurysms. This was an observational retrospective cohort multicentre study. Prospectively collected data from six physician sponsored investigational device exemption studies at US centres were analysed. Patients were treated with fenestrated and branched aortic endografts for pararenal and thoraco-abdominal aortic aneurysms (TAAAs) between 2012 and 2017. Technical success was defined as successful intra-operative catheterisation and stenting of all intended target visceral arteries. Univariable and stratified analyses were performed to identify differences in outcomes between repairs using preloaded and standard devices. There were 564 patients (73% men, mean age 73 ± 8 years) treated for 168 pararenal aortic aneurysms (29.8%), 216 type IV TAAAs (38.3%), and 180 type I - III TAAAs (31.9%). Preloaded grafts (PGs) were used in 387 (68.6%) patients and standard grafts (SGs) in 177 (31.4%). PGs were used preferentially for type IV TAAAs (45% vs. 24%; p < .001), whereas standard devices were used more frequently among patients with type I - III TAAAs (24% vs. 49%; p < .001). The majority of custom made devices were preloaded (95% vs. 21%; p < .001). A total of 2 157 target arteries were incorporated (mean 3.9/patient) utilising 1 469 fenestrations (68.1%), 603 directional branches (27.9%), and 85 double wide scallops (3.9%). Most PGs included fenestrations (80% vs. 43%; p < .001), whereas directional branches were more frequent in standard devices (17% vs. 53%; p < .001). Contrast volume, fluoroscopy time, radiation dose, and operative time were not significantly different between preloaded and standard devices. Upper extremity access was more frequent for PGs (87% vs. 72%; p < .001). Overall technical success was 98.8% and comparable for both preloaded and standard grafts (99.5% vs. 97.2%; p = .022). The 30 day stroke rate was similar for PGs and SGs (2.3% and 1.7%%, respectively). The 30 day mortality rate was 1.9%, and low for both PGs and SGs (0.8% vs. 4.5%; p = .003). Endovascular repair of complex aortic aneurysms is safe and effective. The expanded use of preloaded catheters and wires of fenestrations and directional branches for target artery incorporation is associated with a high technical success and low early mortality.


Blue-enriched white light in the workplace improves self-reported alertness, performance and sleep quality.
Specifications and standards for lighting installations in occupational settings are based on the spectral sensitivity of the classical visual system and do not take into account the recently discovered melanopsin-based, blue-light-sensitive photoreceptive system. The authors investigated the effects of exposure to blue-enriched white light during daytime workhours in an office setting. The experiment was conducted on 104 white-collar workers on two office floors. After baseline assessments under existing lighting conditions, every participant was exposed to two new lighting conditions, each lasting 4 weeks. One consisted of blue-enriched white light (17 000 K) and the other of white light (4000 K). The order was balanced between the floors. Questionnaire and rating scales were used to assess alertness, mood, sleep quality, performance, mental effort, headache and eye strain, and mood throughout the 8-week intervention. Altogether 94 participants [mean age 36.4 (SD 10.2) years] were included in the analysis. Compared with white light (4000 K), blue-enriched white light (17 000 K) improved the subjective measures of alertness (P<0.0001), positive mood (P=0.0001), performance (P<0.0001), evening fatigue (P=0.0001), irritability (P=0.004), concentration (P<0.0001), and eye discomfort (P=0.002). Daytime sleepiness was reduced (P=0.0001), and the quality of subjective nocturnal sleep (P=0.016) was improved under blue-enriched white light. When the participants' expectation about the effect of the light treatments was entered into the analysis as a covariate, significant effects persisted for performance, alertness, evening fatigue, irritability, difficulty focusing, concentrating, and blurred vision. Exposure to blue-enriched white light during daytime workhours improves subjective alertness, performance, and evening fatigue.


CD300f:IL-5 cross-talk inhibits adipose tissue eosinophil homing and subsequent IL-4 production.
Eosinophils and their associated cytokines IL-4 and IL-5 are emerging as central orchestrators of the immune-metabolic axis. Herein, we demonstrate that cross-talk between the Ig-superfamily receptor CD300f and IL-5 is a key checkpoint that modifies the ability of eosinophils to regulate metabolic outcomes. Generation of Il5 Tg /Cd300f -/- mice revealed marked and distinct increases in eosinophil levels and their production of IL-4 in the white and brown adipose tissues. Consequently, Il5 Tg /Cd300f -/- mice had increased alternatively activated macrophage accumulation in the adipose tissue. Cd300f -/- mice displayed age-related accumulation of eosinophils and macrophages in the adipose tissue and decreased adipose tissue weight, which was associated with decreased diet-induced weight gain and insulin resistance. Notably, Il5 Tg /CD300f -/- were protected from diet-induced weight gain and glucose intolerance. These findings highlight the cross-talk between IL-5 receptor and CD300f as a novel pathway regulating adipose tissue eosinophils and offer new entry points for therapeutic intervention for obesity and its complications.


Effects of autologous stromal cells and cytokines on differentiation of equine bone marrow-derived progenitor cells.
OBJECTIVE To develop an in vitro system for differentiation of equine B cells from bone marrow hematopoietic progenitor cells on the basis of protocols for other species. SAMPLE Bone marrow aspirates aseptically obtained from 12 research horses. PROCEDURES Equine bone marrow CD34+ cells were sorted by use of magnetic beads and cultured in medium supplemented with cytokines (recombinant human interleukin-7, equine interleukin-7, stem cell factor, and Fms-like tyrosine kinase-3), murine OP9 stromal cell preconditioned medium, and equine fetal bone marrow mesenchymal stromal cell preconditioned medium. Cells in culture were characterized by use of flow cytometry, immunocytofluorescence microscopy, and quantitative reverse-transcriptase PCR assay. RESULTS For these culture conditions, bone marrow-derived equine CD34+ cells differentiated into CD19+IgM+ B cells that expressed the signature transcription factors early B-cell factor and transcription factor 3. These conditions also supported the concomitant development of autologous stromal cells, and their presence was supportive of B-cell development. CONCLUSIONS AND CLINICAL RELEVANCE Equine B cells were generated from bone marrow aspirates by use of supportive culture conditions. In vitro generation of equine autologous B cells should be of use in studies on regulation of cell differentiation and therapeutic transplantation.


Childhood Stunting and Wasting Following Independence in South Sudan.
South Sudan has experienced ongoing civil and environmental problems since gaining independence in 2011 that may influence childhood nutritional status. To estimate the prevalence of undernutrition among children in South Sudan in 2018 and 2019 compared to the prevalence in 2010. Data on height and weight were collected using a 2-stage stratified sample framework in which households were randomly selected at the county level and nutritional status was calculated for all children under 5 years of age to determine height-for-age, weight-for-height, and weight-for-age Z-scores (HAZ, WHZ, and WAZ) and the prevalence of stunting, wasting, and underweight. Linear and logistic regression analyses were used to determine factors associated with nutritional status and the odds ratio for nutritional outcomes. In 2010, the mean HAZ, WHZ, and WAZ was -0.78, -0.82, and -1.15, respectively, and the prevalence of stunting, wasting, and underweight was 30%, 23%, and 32%, respectively. In 2018 and 2019, the mean HAZ, WHZ, and WAZ was -0.50, -0.70, -0.77 and -0.53, -0.77, -0.76, respectively. The prevalence of stunting, wasting, and underweight in 2018 and 2019 was 17%, 14%, 15% and 16%, 16%, 17%, respectively. Age was negatively associated with all nutritional indices and girls had higher HAZ, WHZ, and WAZ and a lower mid upper arm circumference (P < .01) compared to boys. The risk of poor nutritional outcomes was associated with vaccine status and varied by state of residence. Following independence in 2010, the prevalence of undernutrition in South Sudan decreased, but the risk for undernutrition varied by state and efforts to address food security and health need to ensure equitable access for all children in South Sudan.


Harvesting-resistant estimates of air pollution effects on mortality.
A number of studies have recently shown an association between particle concentrations in outdoor air and daily mortality counts in urban locations. In the public health interpretation of this evidence, a key issue is whether the increased mortality associated with higher pollution levels is restricted to very frail persons for whom life expectancy is short in the absence of pollution. This possibility has been termed the "harvesting hypothesis." We present an approach to estimating the association between pollution and mortality from times series data that is resistant to short-term harvesting. The method is based in the concept that harvesting alone creates associations only at shorter time scales. We use frequency domain log-linear regression to decompose the information about the pollution-mortality association into distinct time scales, and we then create harvesting-resistant estimates by excluding the short-term information that is affected by harvesting. We illustrate the methods with total suspended particles and mortality counts from Philadelphia for 1974-1988. The total suspended particles-mortality association in Philadelphia is inconsistent with the harvesting-only hypothesis, and the harvesting-resistant estimates of the total suspended particles relative risk are actually larger-not smaller-than the ordinary estimates.


The incidence of venous thromboembolism in asymptomatic carriers of a deficiency of antithrombin, protein C, or protein S: a prospective cohort study.
Deficiencies of antithrombin, protein C, and protein S are associated with an increased risk of venous thromboembolism. The objective of this study was to prospectively assess the incidence of venous thromboembolism in nontreated asymptomatic subjects with such a deficiency. We conducted a prospective cohort study in asymptomatic family members of unselected patients who presented with a venous thromboembolic event and who were found to have a deficiency of antithrombin, protein C, or protein S. No anticoagulant prophylaxis was given to the study participants, except during risk periods for venous thromboembolism. All venous thromboembolic events were diagnosed by objective diagnostic tests. A total of 208 individuals with a mean age of 37 years (range, 15 to 79) were included in the study. A total of 611 patient observation years was obtained. Nine venous thromboembolic events occurred, resulting in an annual incidence of 1.5% (95% confidence interval [CI], 0.7 to 2.8) for the 3 deficiencies combined. Five of these events occurred spontaneously, resulting in an annual incidence of spontaneous venous thromboembolism of 0.8% (95% CI, 0.3 to 1.9). For antithrombin, protein C, and protein S deficiencies separately, this figure was 1.6%, 1.0%, and 0.4%, respectively. Thirty-four subjects experienced a total of 40 risk periods during which 4 venous thromboembolic events occurred (10% per risk period). We conclude that the use of continuous anticoagulant prophylaxis seems not warranted in asymptomatic individuals with a deficiency of antithrombin, protein C, or protein S. During risk periods for venous thromboembolism, adequate anticoagulant prophylaxis is necessary.


Epstein-Barr virus-positive diffuse large B-cell lymphoma.
While the World Health Organization included Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) as a provisional entity of a lymphoma occurring in older individuals without any known immunodeficiency in 2008, it has since been recognized that this entity may occur in younger individuals. As a result, the 2016 revision has substituted the modifier "elderly" with "not otherwise specified" (NOS). The NOS highlights that there are more specific entities with neoplastic EBV-positive large B cells such as lymphomatoid granulomatosis. Diagnosis requires that there be no other cause of immunodeficiency and that other more specific entities with neoplastic EBV plus large B cells be excluded. We present the case of an 81-year-old woman hospitalized for generalized weakness, increasing confusion, unexplained weight loss, and intermittent fevers. Examination showed lymphadenopathy, lesions in the liver and small intestine, and a very high EBV viral load. She experienced a rapid demise and at autopsy was found to have EBV+ DLBCL, NOS.


Spatial Proximity and Relative Distribution of Tumor-Infiltrating Lymphocytes and Macrophages Predict Survival in Melanoma.
Tumor microenvironment plays a crucial role in primary cutaneous melanoma (CM) progression. Although the role of tumor-infiltrating lymphocyte (TIL) density has been known for a long time, its spatial distribution and impact with or without tumor-associated macrophages (TAMs) remain controversial. Herein, we investigated spatial proximity between tumor cells and immune cells in 113 primary CM and its correlation with disease-free (DFS) and overall survival (OS). The study cohort included clinical stage II (n = 79) and stage III (n = 34) primary CM with a Breslow thickness of >2 mm (with a median age of 64 years, including 72 men and 41 women). In univariate models, patients with SOX10+ melanoma cells with high proximity to CD8+ TILs in a 20 μm radius showed longer DFS (hazard ratio [HR], 0.58, 95% CI 0.36-0.93, P = .025) and OS (HR, 0.55, 95% CI 0.32-0.92, P = .023). Furthermore, at multivariate combined analysis, patients with SOX10+ melanoma cells with high proximity to CD8+ TILs or low proximity to CD163+ TAMs in a 20 μm radius showed an increased OS (aHR, 0.37, 95% CI 0.14-0.96, P = .04) compared with melanoma patients with low proximity to CD8+ TILs or high proximity to CD163+ TAMs. In a subgroup analysis including 92 patients, a significant negative impact on DFS (aHR 4.49, 95% CI 1.73-11.64, P = .002) and OS (aHR 3.97, 95% CI 1.37-11.49, P = .01) was observed in sentinel lymph node (SLN)-negative patients with a high proximity of CD163+ TAMs to CD8+ TILs. These findings could help identify high-risk patients in the context of thick melanoma and a negative SLN. Our study suggests the importance of quantifying not only the density of immune cells but also the individual and combined relative spatial distributions of tumor cells and immune cells for clinical outcomes in SLN-negative primary CM patients.


Congress of Neurological Surgeons Systematic Review and Evidence-Based Guideline for Pretreatment Endocrine Evaluation of Patients With Nonfunctioning Pituitary Adenomas.
Nonfunctioning pituitary adenomas (NFPAs) are among the most common pituitary lesions and may present with hypopituitarism and/or hyperprolactinemia. To review the existing literature as it pertains to preoperative endocrine assessment in the workup for NFPAs. A systematic review methodology was utilized to identify and screen articles assessing the role and results of preoperative laboratory assessment in patients with NFPAs. The prevalence of individual pituitary hormonal axis deficiencies was reviewed. Twenty-nine studies met inclusion criteria for analysis. No class I evidence was available, and all studies met criteria for class II evidence. Baseline serum laboratory assessment showed a prevalence of overall hypopituitarism in 37% to 85% of patients. The most common hormonal axis deficiency was growth hormone deficiency, prevalent in 61% to 100% of patients. The next most common deficit was hypogonadism, seen in 36% to 95% of patients. Adrenal insufficiency was diagnosed in 17% to 62% of patients. Finally, hypothyroidism was seen in 8% to 81% of patients. Hyperprolactinemia was seen in 25% to 65% of patients, with a mean level of 39 ng/mL and with a minority of patients exceeding a serum prolactin level of 200 ng/mL. No evidence supporting routine biomarker testing (eg, α-subunit or chromogranin A) or genetic testing in patients with sporadic NFPAs was available. Despite a paucity of class I evidence, multiple retrospective studies have demonstrated a high prevalence of hypopituitarism in patients with NFPAs. Routine endocrine analysis of all anterior pituitary axes to assess for hypopituitarism is recommended, with prolactin and insulin-like growth factor 1 evaluation also valuable to assess for hypersecretion states that might not be clinically suspected. The full guidelines document for this chapter can be located at https://www.cns.org/guidelines/guidelines-management-patients-non-functioning-pituitary-adenomas/Chapter_3. GH, growth hormoneIGF-1, insulin-like growth factor 1NFPA, nonfunctioning pituitary adenoma.


Theoretical models of directional proton molecular transport.
The important topic of proton transport through molecular wires is usually associated with the Grotthuss mechanism. In this paper we propose an alternative conductor based on chains of lone pairs. B3LYP/6-31+G** and PW91 DFT calculations on model compounds (1,2,3,4-tetrasubstituted benzenes) show that these compounds could play the role of proton conductors.


[Leukocyte lipids in mature cell leukemia].
The total lipid content and the lipid pattern of normal and leukemic leukocytes showed no significant difference except the fact, that lymphocytes of chronic lymphatic leukemia have a lower content of triglycerides and sphingomyeline. Lecithin of leukemic lymphocytes contained more palmitinic acid and oleic acid and a lower level of stearic acid than the same fraction in normal lymphocytes. The lipid pattern of normal and leukemic granulocytes was identical.


Walking activities and wear of prostheses.
A study of the walking activities of 243 individuals was carried out. The individuals came from four different occupations and had an age range of 17-83 years. The survey carried out in this investigation showed surprisingly little correlation between variables such as age, height, and weight of individuals and their speed of walking, length of stride, or distance walked. Correlation matrices were obtained for the whole sample and then for each sex, showing similar trends. The most significant correlation was between the height of an individual and the length of the stride, and there was a lesser correlation between age and the number of steps walked by an individual per day. Further statistical analyses showed that males in manual occupations walked most and those in sedentary occupations walked least. On the other hand, in the female groups housewives seemed to walk least and those in technical occupations walked most. The average number of steps walked per day by a male individual for the whole sample was 9537; that for females was 9839. The corresponding distances walked per day were 6.7 and 6.5 km. The differences were not statistically significant. Predictions of wear of prosthetic components made of ultrahigh molecular weight polyethylene were made on the basis of the above data and other variables affecting wear, such as the weight of the subject and the area available for contact during walking. Charts have been constructed of the penetration of the metallic component into the plastic one for both hip knee prostheses, thus enabling predictions of the wear of the plastic components of these two most widely used prostheses. Owing to the wide ranging values of the variables used in making the predictions of wear, these latter should be regarded only as 'safe' first estimates.


12-O-tetradecanoylphorbol-13-acetate (TPA) increases murine intestinal crypt stem cell survival following radiation injury.
Radiation enteropathy is a common complication in cancer patients following radiation therapy. Thus, there is a need for agents that can protect the intestinal epithelium against radiation. 12-O-tetradecanoylphorbol-13-acetate (TPA) has been shown to induce differentiation and/or apoptosis in multiple cell lines and primary cells. In the current report, we studied the function of TPA in radiation induced enteropathy in cultured rat intestinal epithelial cell line IEC-6 after ionizing radiation (IR) and in mice after high dose total-body gamma-IR (TBI). In IEC-6 cells, there were reduced apoptosis and cell cycle arrest in TPA treated cells after IR. We detected a four-fold increase in crypt cell survival and a two-fold increase in animal survival post TBI in TPA treated mice. The beneficial effects of TPA were accompanied by upregulation of stem cells markers and higher level of proteins that are involved in PKC signaling pathway. In addition, TPA also decreased the TBI-augmented levels of the DNA damage indicators. The effects were only observed when TPA was given before irradiation. These results suggest that TPA has the ability to modulate intestinal crypt stem cells survival and this may represent a promising countermeasure against radiation induced enteropathy.


[Assessment instruments for a Health-Related Quality of Life in diabetes mellitus].
The assessment of Health-Related Quality of Life (HRQoL) has been increasingly used to measure the overall impact of diseases in people's life. Diabetes mellitus (DM) is a chronic disease associated with high morbidity, mortality, and HRQoL impairment in patients. In longitudinal studies, the psychosocial impact of DM predicts mortality. The objective of this review is to describe and to analyze the main instruments used for the HRQoL evaluation in patients with DM. Generic instruments such, as the Quality of Well-Being Scale (QWB), Medical Outcomes Study 36-item Short-Form Health Survey (SF-36), EuroQol (EQ-5D) and specific instruments as the Diabetes Care Profile (DCP), Diabetes Quality of Life Measure (DQOL), Diabetes Impact Measurement Scales (DIMS), Appraisal of Diabetes Scale (ADS), Audit of Diabetes-Dependent Quality of Life (ADDQoL), Diabetes Health Profile (DHP-1 and DHP-18), Questionnaire on Stress in Patients with Diabetes-Revised (QSD-R), Well-Being Enquiry goes Diabetics (WED), Diabetes-Specific Quality-of-life Scale (DSQOLS), Diabetes 39 (D-39) Problems Areas in Diabetes (PAID) were analyzed. PAID is the only translated and validated instrument available in Brazil. The generic and specific instruments have their stregths and shortcomings for evaluation of HRQL in patients with DM. The combined use of both generic (such as the SF-36) and specific (such as the PAID) appears to be a consistent way to evaluate HRQoL as a construct in Brazilian patients with DM. The present article reviews a variety of instruments and emphasizes the urgent need for validation studies of such instruments to be used in Brazilian subjects with DM.


Parents' presence and participation in medical rounds in 11 European neonatal units.
Parents' involvement during hospital care is beneficial for preterm infants and their parents. Although parents are encouraged to be present in many neonatal intensive care units (NICUs), little is known about their role during medical rounds. To study parents' presence in the NICU, the degree of parents' participation during medical rounds, and to identify underlying factors for participation. A prospective study was performed in 11 neonatal units in six European countries including parents of preterm infants born before 35 gestational weeks. Parents' presence and the degree of participation (7-point Likert scale) during medical rounds were asked using a text-message question sent to the mobile phone of each parent separately. A total of 241 families were included in the study; mothers responded to 630 and fathers to 474 text-message questions, respectively. In studied units, mothers were present during medical rounds on 62.5% to 91% and fathers 30.8% to 77.8% of the days. The degree of mothers' and fathers' participation also varied between units (p < 0.001 and p = 0.022, respectively). In multivariate analysis, parents' presence increased by increasing gestational age (p = 0.010), fathers' education (p = 0.009), and by the policy in the unit to invite parents to medical rounds (p = 0.036). The background characteristics did not explain the degree of participation. There is significant variation between neonatal units in how they include parents in medical rounds. Only few background characteristics explained the differences suggesting that unit culture plays a major role in welcoming parents to participate.


Complete genome sequence of the fish pathogen Flavobacterium psychrophilum ATCC 49418(T.).
Flavobacterium psychrophilum is the causative agent of bacterial cold water disease and rainbow trout fry mortality syndrome in salmonid fishes and is associated with significant losses in the aquaculture industry. The virulence factors and molecular mechanisms of pathogenesis of F. psychrophilum are poorly understood. Moreover, at the present time, there are no effective vaccines and control using antimicrobial agents is problematic due to growing antimicrobial resistance and the fact that sick fish don't eat. In the hopes of identifying vaccine and therapeutic targets, we sequenced the genome of the type strain ATCC 49418 which was isolated from the kidney of a Coho salmon (Oncorhychus kisutch) in Washington State (U.S.A.) in 1989. The genome is 2,715,909 bp with a G+C content of 32.75%. It contains 6 rRNA operons, 49 tRNA genes, and is predicted to encode 2,329 proteins.


[Rapidly progressing glomerulonephritis with mesangial IgA deposits in sarcoidosis].
This report concerns a patient with sarcoidosis who developed acute renal failure due to rapidly progressive glomerulonephritis. Renal biopsy revealed severe endo and extracapillary proliferation with diffuse mesangial deposits of IgA, C3 and beta 1-H on immunofluorescence. The possibility of a common immunopathogenic pathway between sarcoidosis and IgA nephritis is briefly discussed.


Genetic isolation between two recently diverged populations of a symbiotic fungus.
Fungi are an omnipresent and highly diverse group of organisms, making up a significant part of eukaryotic diversity. Little is currently known about the drivers of fungal population differentiation and subsequent divergence of species, particularly in symbiotic, mycorrhizal fungi. Here, we investigate the population structure and environmental adaptation in Suillus brevipes (Peck) Kuntze, a wind-dispersed soil fungus that is symbiotic with pine trees. We assembled and annotated the reference genome for Su. brevipes and resequenced the whole genomes of 28 individuals from coastal and montane sites in California. We detected two clearly delineated coast and mountain populations with very low divergence. Genomic divergence was restricted to few regions, including a region of extreme divergence containing a gene encoding for a membrane Na(+) /H(+) exchanger known for enhancing salt tolerance in plants and yeast. Our results are consistent with a very recent split between the montane and coastal Su. brevipes populations, with few small genomic regions under positive selection and a pattern of dispersal and/or establishment limitation. Furthermore, we identify a putatively adaptive gene that motivates further functional analyses to link genotypes and phenotypes and shed light on the genetic basis of adaptive traits.


[Complications associated with soft hydrophilic contact lenses].
The authors report 89 cases of ocular complications associated with soft hydrophilic contact lenses, including corneal abrasions, corneal ulcers, punctate corneal epithelial erosions, aphakic corneal edema, and superior limbal keratoconjunctivitis. Soft contact lenses interfere with the integrity and function of the tear film, so that corneal metabolism is affected, predisposing the cornea to infections by Pseudomonas, Acanthamoebia, fungus and virus. The incidence of the complications is high; therefore, due attention should be devoted to their prevention and treatment.


Juxtamembrane shedding of Plasmodium falciparum AMA1 is sequence independent and essential, and helps evade invasion-inhibitory antibodies.
The malarial life cycle involves repeated rounds of intraerythrocytic replication interspersed by host cell rupture which releases merozoites that rapidly invade fresh erythrocytes. Apical membrane antigen-1 (AMA1) is a merozoite protein that plays a critical role in invasion. Antibodies against AMA1 prevent invasion and can protect against malaria in vivo, so AMA1 is of interest as a malaria vaccine candidate. AMA1 is efficiently shed from the invading parasite surface, predominantly through juxtamembrane cleavage by a membrane-bound protease called SUB2, but also by limited intramembrane cleavage. We have investigated the structural requirements for shedding of Plasmodium falciparum AMA1 (PfAMA1), and the consequences of its inhibition. Mutagenesis of the intramembrane cleavage site by targeted homologous recombination abolished intramembrane cleavage with no effect on parasite viability in vitro. Examination of PfSUB2-mediated shedding of episomally-expressed PfAMA1 revealed that the position of cleavage is determined primarily by its distance from the parasite membrane. Certain mutations at the PfSUB2 cleavage site block shedding, and parasites expressing these non-cleavable forms of PfAMA1 on a background of expression of the wild type gene invade and replicate normally in vitro. The non-cleavable PfAMA1 is also functional in invasion. However - in contrast to the intramembrane cleavage site - mutations that block PfSUB2-mediated shedding could not be stably introduced into the genomic pfama1 locus, indicating that some shedding of PfAMA1 by PfSUB2 is essential. Remarkably, parasites expressing shedding-resistant forms of PfAMA1 exhibit enhanced sensitivity to antibody-mediated inhibition of invasion. Drugs that inhibit PfSUB2 activity should block parasite replication and may also enhance the efficacy of vaccines based on AMA1 and other merozoite surface proteins.


BRCA1 requirement for the fidelity of plasmid DNA double-strand break repair in cultured breast epithelial cells.
The tumor suppressor breast cancer susceptibility protein 1 (BRCA1) protects our cells from genomic instability in part by facilitating the efficient repair of DNA double-strand breaks (DSBs). BRCA1 promotes the error-free repair of DSBs through homologous recombination and is also implicated in the regulation of nonhomologous end joining (NHEJ) repair fidelity. Here, we investigate the role of BRCA1 in NHEJ repair mutagenesis following a DSB. We examined the frequency of microhomology-mediated end joining (MMEJ) and the fidelity of DSB repair relative to BRCA1 protein levels in both control and tumorigenic breast epithelial cells. In addition to altered BRCA1 protein levels, we tested the effects of cellular exposure to mirin, an inhibitor of meiotic recombination enzyme 11 (Mre11) 3'-5'-exonuclease activity. Knockdown or loss of BRCA1 protein resulted in an increased frequency of overall plasmid DNA mutagenesis and MMEJ following a DSB. Inhibition of Mre11-exonuclease activity with mirin significantly decreased the occurrence of MMEJ, but did not considerably affect the overall mutagenic frequency of plasmid DSB repair. The results suggest that BRCA1 protects DNA from mutagenesis during nonhomologous DSB repair in plasmid-based assays. The increased frequency of DSB mutagenesis and MMEJ repair in the absence of BRCA1 suggests a potential mechanism for carcinogenesis.


Characterization of microalgal carotenoids by mass spectrometry and their bioavailability and antioxidant properties elucidated in rat model.
Of the total carotenoids in respective algal samples, beta-carotene in Spirulina platensis was 69.5%, astaxanthin and its esters in Haematococcus pluvialis was 81.38%, and lutein in Botryococcus braunii was 74.6%. The carotenoids were characterized by mass spectrometry. A time-course study of carotenoids in rats after administration of microalgal biomass showed peak levels in plasma, liver, and eyes at 2, 4, and 6 h, respectively. Beta-carotene accumulation in Spirulina-fed rats was maximum in eye tissues at 6 h. Similarly, levels of astaxanthin and lutein in Haematococcus- and Botryococcus-fed rats were also maximal in eye tissues. Astaxanthin from H. pluvialis showed better bioavailability than beta-carotene and lutein. The antioxidant enzymes, catalase, superoxide dismutase, peroxidase, and TBARS were significantly high in plasma at 2 h and in liver at 4 h, evidently offering protection from free radicals. This study implies that microalgae can be a good source of carotenoids of high bioavailability and nutraceutical value.


The suppression and promotion of DNA charge inversion by mixing counterions.
In the preset study, we report the suppression and promotion of DNA charge inversion by mixing a quadrivalent counterion (spermine) with mono-, di- and trivalent counterions by dynamic light scattering (DLS) and single molecule electrophoresis (SME) methods. We find that the electrophoretic mobility of DNA in spermine solution decreases in the presence of monovalent sodium ions and divalent magnesium ions. It means that the charge neutralization of DNA by the quadrivalent counterion is suppressed when adding extra mono- or divalent counterions. More specifically, at a high concentration of spermine, the positive mobility can switch back to a negative value by adding mono- and divalent counterions. Thus, charge neutralization and inversion of DNA by quadrivalent counterions is suppressed in the mono- and divalent ion solution. However, the scenario changes dramatically when we add trivalent ions into the solution of DNA and spermine. In this case, the charge neutralization and inversion of DNA is promoted rather than suppressed by mixing with trivalent ions. The negative electrophoretic mobility can be promoted to a positive value, which corresponds to the charge inversion, by trivalent counterions. Thus trivalent and quadrivalent counterions work cooperatively in DNA charge neutralization and inversion. This promotion also occurs when highly positively charged chitosan is introduced into the solution. We explain the observation by the counterion complexation that is related to DNA condensation, which is supported by the images of atomic force microscopy (AFM).


The effect of CO2 laser pulse repetition rate on tissue ablation rate and thermal damage.
The ablation rate and thermal damage in skin produced by a superpulsed CO2 laser operating at pulse repetition rates between 1 and 900 Hz was measured. When delivering a fixed number of pulses (20 or 30) of equal energy, a 55-60% increase in the amount of tissue ablated was observed when the pulse repetition rate rose from 10 to 200 Hz. At pulse repetition rates greater than 200 Hz no further increase was seen. Under identical conditions, an 80% increase in the zone of thermal damage was observed when the pulse repetition rate was increased from 1 to 60 Hz. The large increases in tissue ablation and tissue damage may indicate the existence of a layer of mixed-phase (i.e., liquid and vapor) or metastable liquid which can store significant amounts of thermal energy between pulses. The data suggest that CO2 lasers should be operated at relatively low repetition rates for optimal performance.


Subject characteristics as predictors of self-change in smoking.
Subject characteristics served as predictors of smokers' success or failure in their self-change efforts at smoking cessation over a 6-month period. Seven hundred and three adult subjects represented five stages of change based on the transtheoretical model: precontemplation, contemplation, action, maintenance, and relapse. Step-wise multiple discriminant functions were used to predict movement across the stages. Health problems, problem duration, daily cigarettes, previous attempts to quit, and smoking for pleasure predicted movement through various stages of change. The stronger the smoking habit, the less apt the smoker is to quit or maintain a nonsmoking status. Smokers with higher incomes and more years of education are more likely than persons of lower socioeconomic levels to be successful in their self-change efforts. The advantages of process-oriented rather than static variables for predicting self-change are discussed.


Time trends in emergency department use among adults with intellectual and developmental disabilities.
Emergency Department (ED) visits are common among adults with intellectual and developmental disabilities (IDD). However, little is known about how ED use has varied over time in this population, or how it has been affected by recent Medicaid policy changes. To examine temporal trends in ED use among adult Medicaid members with IDD in eight states that ranged in the extent to which they had implemented state-level Medicaid policy changes that might affect ED use. We conducted repeated cross-sectional analyses of 2010-2016 Medicaid claims data. Quarterly analyses included adults ages 18-64 years with IDD (identified by diagnosis codes) who were continuously enrolled in Medicaid for the past 12 months. We assessed change in number of ED visits per 1000 member months from 2010 to 2016 overall and interacted with state level policy changes such as Medicaid expansion. States with no Medicaid expansion experienced an increase in ED visits (linear trend coefficient: 1.13, p < 0.01), while states operating expansion via waiver had a much smaller (non-significant) increase, and states with ACA-governed expansion had a decrease in ED visits (linear trend coefficient: 1.17, p < 0.01). Other policy changes had limited or no association with ED visits. Medicaid expansion was associated with modest reduction or limited increase in ED visits compared to no expansion. We found no consistent decrease in ED visits in association with other Medicaid policy changes.


NGS analysis in Marfan syndrome spectrum: Combination of rare and common genetic variants to improve genotype-phenotype correlation analysis.
The diagnosis of Marfan spectrum includes a large number of clinical criteria. Although the identification of pathogenic variants contributes to the diagnostic process, its value to the prediction of clinical outcomes is still limited. An important novelty of the present study is represented by the statistical approach adopted to investigate genotype-phenotype correlation. The analysis has been improved considering the extended genetic information obtained by Next Generation Sequencing (NGS) and combining the effects of both rare and common genetic variants in an inclusive model. To this aim a cohort of 181 patients were analyzed with a NGS panel including 11 genes associated with Marfan spectrum. The genotype-phenotype correlation was also investigated considering the possibility to predict presence of a pathological mutation in Marfan syndrome (MFS) main genes based only on the analysis of phenotypic traits. Results obtained indicate that information about clinical traits can be summarized in a new variable that resulted significantly associated with the probability to find a pathological mutation in MFS main genes. This is important since the choice of the genetic test is often influenced by the phenotypic characterization of patients. Moreover, both rare and common variants were found to significantly contribute to clinical spectrum and their combination allowed to increase the percentage of phenotype variability that could be explained based on genetic factors. Results highlight the opportunity to take advantage of the overall genetic information obtained by NGS data to have a better clinical classification of patients.


Socioeconomic inequalities in prognostic markers of non-Hodgkin lymphoma: analysis of a national clinical database.
The survival of non-Hodgkin lymphoma patients strongly depends on a range of prognostic factors. This registry-based clinical cohort study investigates the relation between socioeconomic position and prognostic markers in 6234 persons included in a national clinical database in 2000-2008, Denmark. Several measures of individual socioeconomic position were achieved from Statistics Denmark. The risk of being diagnosed with advanced disease, as expressed by the six prognostic markers (Ann Arbor stage III or IV, more than one extranodal lesion, elevated serum lactate dehydrogenase (LDH), performance status of two or more, presence of B symptoms and International Prognostic Index (IPI) of two or more), increased with decreasing level of education, in patients living alone, and in men. For instance, a significant decrease in the odds of being diagnosed with elevated LDH (p=0.02), high performance status (p=0.004), high IPI score (p=0.004) and B symptoms (p=0.02) was seen with higher level of education, whereas high stage of disease was significantly less likely in the higher educated (odds ratio [OR]=0.85 (0.74-0.99)). The difference in risk seemed not to be mediated by differences in histological subgroups reflecting aggressiveness of disease among the social groups. One of the most likely mechanisms of the social difference is longer delay in those with low socioeconomic position. The findings of social inequality in prognostic markers in non-Hodgkin lymphoma (NHL) patients could already be implemented in the clinical practice if general practitioners (GP's) and physicians on hospitals paid special attention to patients with low educational level and unspecific symptoms.


[Nonspecific gamma-esterase in the tissues in endometrial cancer].
Malignization of endometrium is accompanied by an increase in the production of nonspecific tissue gamma-esterase (NTE). NTE content in uterine tissues and mucus of patients with cancer of endometrium correlates with a degree of differentiation, spreading and depth of tumour cell invasion into the myometrium. The synthesis of NTE occurs in epithelial cells of the endometrium glands which secrete it into the uterus cavity.


Serum thyroglobulin as a risk factor for thyroid carcinoma.
Samples from a biological serum bank taken up to 23 years prior to diagnosis of thyroid carcinoma were analysed for human thyroglobulin, thyroid-stimulating hormone and thyroxin. After exclusions, the final study material consisted of 59 cases of papillary and follicular carcinomas. These cases were compared with 164 controls, matched for sex, age and time of sample taking. The most interesting finding was that concentrations of thyroglobulin in serum were abnormally elevated in cases compared with controls, equal to or above 30 microg/L, with odds ratio 7.0 (CI 3.1-15.7). This elevation of serum thyroglobulin occurred in 44% of the carcinoma cases. Sensitivity was around 50 for measurements taken up to 15 years prior to diagnosis, but 21 when the interval was over 15 years. Specificity was 89. No differences were found between cases and controls in values for thyroid-stimulating hormone and thyroxin.


First principles analysis of surface dependent segregation in bimetallic alloys.
Stability is an important aspect of alloys, and proposed alloys may be unstable due to unfavorable atomic interactions. Segregation of an alloy may occur preferentially at specific exposed surfaces, which could affect the alloy's structure since certain surfaces may become enriched in certain elements. Using density functional theory (DFT), we modeled surface segregation in bimetallic alloys involving all transition metals doped in Pt, Pd, Ir, and Rh. We not only modeled common (111) surfaces of such alloys, but we also modeled (100), (110), and (210) facets of such alloys. Segregation is more preferred for early and late transition metals, with middle transition metals being most stable within the parent metal. We find these general trends in segregation energies for the parent metals: Pt > Rh > Pd > Ir. A comparison of different surfaces suggests no consistent trends across the different parent hosts, but segregation energies can vary up to 2 eV depending on the exposed surface. We also developed a statistical model to predict surface-dependent segregation energies. Our model is able to distinguish segregation at different surfaces (as opposed to generic segregation common in previous models), and agrees well with the DFT data. The present study provides valuable information about surface-dependent segregation and helps explain why certain alloy structures occur (e.g. core-shell).


[Investigation of the Relationship Between IL28B Polymorphisms and Plasma IL28B Levels in Patients with Chronic Hepatitis B or C].
Some single nucleotide polymorphisms (SNPs) of the gene encoding interleukin 28B (IL28B) may increase susceptibility to infection and chronicity in humans with hepatitis B and C viruses. In our study, we aimed to investigate the prevalence of rs12979860, rs8099917 and rs12980275 SNPs in IL28B in patients with hepatitis B (HBV) or hepatitis C Virus (HCV) infection and to determine the relationship of these polymorphisms with plasma IL28B levels. For this purpose, 64 HBV-infected and 66 HCV-infected patients and 70 healthy individuals were included in the study. The SNPs were investigated by real time PCR (Polimerase Chain Reaction, Rt-PCR) using TaqMan SNP Genotyping Assay. The plasma levels of IL28B were detected by 'Enzyme Linked Immunosorbent Assay (ELISA)'. The frequencies of the rs12980275AG genotype and G allele (p= 0.003 and p= 0.04, respectively), and the rs12979860CT genotype and T allele (p= 0.01 and p= 0.04, respectively) were lower in HBV-infected patients. In HCV-infected patients, the rs8099917TG genotype and G allele frequencies (p= 0.04) were higher and the TGG haplotype showed a statistically significant difference (p= 0.04). The mean of IL28B plasma levels were higher in the control group than the HBV or HCV-infected patient groups (p= 0.001 and p= 0.01, respectively). However, HBV-infected patients with the rs12980275AG genotype showed a significant difference in plasma IL28B levels compared to the other genotypes (p= 0.0001) and these patients had lower viral loads (<105 IU/ml). According to the results of the study, it can be stated that rs12979860CT and rs12980275AG genotypes may play a role in preventing the chronicity of HBV infection, while rs8099917TG genotype may contribute to the transformation of HCV infection into chronic infection. In this study, it was observed that the presence of the G allele for the rs8099917 polymorphism could be evaluated as a risk allele for chronic HCV infection and that the TGG haplotype could have a strong predictive effect on increasing susceptibility to chronic HCV infection. It is recommended to evaluate the genotypic distribution of IL28B before treatment because of its prognostic significance in HBV or HCV infected patients. In HBV infection, the rs12980275AG genotype which is thought to have a protective effect by limiting viral replication with increased plasma IL28B levels, can be used as a good prognostic factor. These polymorphisms could be used as biomarkers to predict the clinical consequences of the patients infected with HBV or HCV, to take precautions to prevent the chronicity of the infection and its complications, and to develop new molecular targeted therapies with further research.


Neuroprotective properties of tianeptine: interactions with cytokines.
Tianeptine is an antidepressant with proven clinical efficacy and effects on hippocampal plasticity. Hypoxia increased lactate dehydrogenase (LDH) release from cortical neuronal cultures, and tianeptine (1, 10 and 100 microM) inhibited LDH release as efficiently as the N-methyl-D-aspartate (NMDA) antagonist, MK-801. However, tianeptine did not block apoptosis in cultured cortical neurones caused by NMDA, but reduced apoptosis when interleukin-1beta (IL-1beta) was included with NMDA. In 5-day old mice, intracerebral injection of ibotenate induced reproducible lesions in cortex and white matter. Lesion size was markedly reduced by co-administration of MK-801 (1 mg/kg i.p.) but neither by tianeptine or its enantiomers administered acutely (1, 3 or 10 mg/kg i.p.) nor by tianeptine administered chronically (10 mg/kg i.p. for 5 days). Chronic administration of IL-1beta (10 ng/kg i.p. for 5 days) prior to ibotenate injection exacerbated lesion size in cortex and white matter, and this exacerbation was prevented by chronic pre-treatment with tianeptine (10 mg/kg i.p.) or by acute administration of tianeptine (10 mg/kg i.p.) concomitantly with ibotenate. Thus tianeptine has neuroprotective effects against hypoxia in tissue culture and against the deleterious effects of cytokines in cortex and white matter, but not against NMDA receptor-mediated excitotoxicity.


Extreme diversity, conservation, and convergence of spider silk fibroin sequences.
Spiders (Araneae) spin high-performance silks from liquid fibroin proteins. Fibroin sequences from basal spider lineages reveal mosaics of amino acid motifs that differ radically from previously described spider silk sequences. The silk fibers of Araneae are constructed from many protein designs. Yet, the repetitive sequences of fibroins from orb-weaving spiders have been maintained, presumably by stabilizing selection, over 125 million years of evolutionary history. The retention of these conserved motifs since the Mesozoic and their convergent evolution in other structural superproteins imply that these sequences are central to understanding the exceptional mechanical properties of orb weaver silks.


Absence of PCB Hot Spot Effect in Walleye Sander vitreus from Lower Green Bay of Lake Michigan.
Under certain conditions, polychlorinated biphenyl (PCB) concentration in individuals of one sex of an adult fish population may exceed that of the other sex by more than a factor of two. This phenomenon, known as the PCB hot spot effect, has been postulated to be contingent upon the following two conditions: (1) presence of a PCB hot spot in the bottom sediments of the aquatic ecosystem, such that prey PCB concentrations in the hot spot region are substantially higher than prey PCB concentrations in locations distant from the hot spot, and (2) habitat use varying between the sexes, such that individuals of one sex inhabit the hot spot region to a considerably greater degree than individuals of the other sex. To test whether PCB concentrations in walleye Sander vitreus from lower Green Bay of Lake Michigan displayed a PCB hot spot effect, whole-fish PCB concentrations were determined in ten female and ten male adult walleye from the population spawning in the Fox River, the main tributary to lower Green Bay. In addition, mark-recapture data for the Fox River walleye population were analyzed to determine differences in spatial distributions between the sexes. Results revealed that the ratio of mean PCB concentration in males to mean PCB concentration in females was only 1.13, indicating the absence of a PCB hot spot effect. This result was corroborated by the mark-recapture data analysis, which showed no significant difference in habitat use between the sexes. Thus, although condition 1 was met, condition 2 was not met. Consequently, the PCB hot spot effect was not observed in the Fox River walleye population. Lack of a significant difference in PCB congener distributions between the sexes further corroborated our conclusions.


Atypical Locations of Renal Cell Carcinoma Metastases to the Pancreas and Duodenum.
Pancreatic tumors account for a small percentage of all malignancies. Most of them are primary and originate from cells of the exocrine pancreas. The remaining primary changes are neuroendocrine tumors. The pancreas may also be a target of metastatic lesions. The most common cancer that metastasizes to the pancreas is renal cell carcinoma. The paper presents two cases of rare metastases to organs of the abdominal cavity: the first patient treated surgically due to two metastatic lesions of renal cell carcinoma in the pancreas, diagnosed many years after radical nephrectomy. The second case of high gastrointestinal obstruction in the course of metastasis of renal cell carcinoma to the duodenum. The first patient underwent distal laparotomic resection of the pancreas. The second patient underwent resection of the duodenum and the first jejunal loop with side-to-side duodeno-jejunal anastomosis. Both patients remain under oncological supervision. Patients after radical nephrectomy due to renal cell carcinoma require long-term systematic monitoring. Due to the anatomical position of the pancreas and duodenum as well as the number, location and size of metastatic lesions, the course of the disease may be initially asymptomatic or oligosymptomatic. Aggressive surgical treatment of pancreatic metastases creates opportunities for long-term survival.


Yield of diagnostic testing in evaluating etiology and end organ effects of pediatric hypertension.
Current recommendations regarding the utility of diagnostic investigations for pediatric hypertension are based on limited evidence, leading to wide practice variation. The objective of this study was to characterize the cohort of children that may benefit from secondary hypertension workup, and determine the diagnostic yield of investigations. This was a single-center, retrospective cohort study of 169 children aged 1-18 years referred between 2000 and 2015, to a tertiary pediatric nephrology center in Canada, for evaluation of hypertension. The number of investigations completed, abnormal findings, and diagnostic findings that helped establish hypertension etiology was determined. 56 children were diagnosed with primary and 72 children with secondary hypertension in the outpatient setting. Secondary hypertension was predominant at all ages except for obese adolescents ≥ 12 years. Half of children with traditional risk factors for primary hypertension, including obesity, were diagnosed with secondary hypertension. Kidney ultrasound had the highest yield of diagnostic results (19.8%), with no difference in yield between age groups (P = 0.19). Lipid profile had a high yield of abnormal results (25.4%) as part of cardiovascular risk assessment but was only abnormal in overweight/obese children. Echocardiogram had a high yield for identification of target-organ effects in hypertensive children (33.3%). A simplified secondary hypertension workup should be considered for all hypertensive children and adolescents. High yield investigations include a kidney ultrasound, lipid profile for overweight/obese children, and echocardiograms for assessment of target-organ damage. Further testing could be considered based on results of initial investigations for the most cost-effective management. A higher resolution version of the Graphical abstract is available as Supplementary information.


Cell cycle-mediated regulation of plant infection by the rice blast fungus.
To gain entry to plants, many pathogenic fungi develop specialized infection structures called appressoria. Here, we demonstrate that appressorium morphogenesis in the rice blast fungus Magnaporthe oryzae is tightly regulated by the cell cycle. Shortly after a fungus spore lands on the rice (Oryza sativa) leaf surface, a single round of mitosis always occurs in the germ tube. We found that initiation of infection structure development is regulated by a DNA replication-dependent checkpoint. Genetic intervention in DNA synthesis, by conditional mutation of the Never-in-Mitosis 1 gene, prevented germ tubes from developing nascent infection structures. Cellular differentiation of appressoria, however, required entry into mitosis because nimA temperature-sensitive mutants, blocked at mitotic entry, were unable to develop functional appressoria. Arresting the cell cycle after mitotic entry, by conditional inactivation of the Blocked-in-Mitosis 1 gene or expression of stabilized cyclinB-encoding alleles, did not impair appressorium differentiation, but instead prevented these cells from invading plant tissue. When considered together, these data suggest that appressorium-mediated plant infection is coordinated by three distinct cell cycle checkpoints that are necessary for establishment of plant disease.


Optimizing vitamin D naming conventions in computerized order entry to support high-value care.
To reduce wasteful ordering of rare 1,25-OH vitamin D lab tests through use of a noninterruptive decision support tool. We conducted a time series quality improvement study at 2 academic hospitals. The titles of vitamin D tests and the order in which they appeared in search results were changed to reflect the purpose and rarity of the tests. We used interruptive time series analyses to evaluate the changes we made. The estimated number of monthly tests ordered at the 2 hospitals increased, by 24.8 and 14.2, following the introduction of computerized provider order entry (CPOE) (both P < .001). When we changed the titles of the tests, the estimated number of monthly tests decreased at the 2 hospitals, by 22.1 and 11.3 (both P < .001). The search order did not affect test utilization. Changing catalog names in CPOE systems for infrequently used tests can reduce unintentional overuse. Users may prefer this to interruptive or restrictive interventions. CPOE vendors and users should refine interfaces by incorporating human factors engineering. Health care institutions should monitor test utilization for unintended changes after CPOE implementation.


Interaction between water and poly(vinylpyrrolidone) containing polyethylene glycol.
Information on the interaction between water and polymers is indispensable for manufacturing solid dispersion of a drug by hot-melt extrusion because this interaction affects various properties of the water-polymer mixtures, such as their viscoelastic properties. In this study, poly(vinylpyrrolidone) K30 (PVP) containing 0%, 10%, and 20% poly(ethylene glycol) 400 (PEG) was used as model amorphous polymers. The interaction of water with these polymers was assessed by the evaluation of the glass transition temperature (Tg), the point on the isotherm corresponding to the weight of sorbed water required to form a complete monolayer on the solid surface (apparent Wm), and the maximal amount of nonfreezing water, which were measured by differential scanning calorimetry and water sorption isotherms. In all of the systems with a water content below a certain water fraction (0.1 for PVP, 0.12 for PVP-PEG 10%, and 0.16 for PVP-PEG 20%), the Tg values were successfully predicted using theoretical equations, whereas the experimental Tg values were higher than predicted for those with a water content above these water fraction levels. In addition, these values of water fraction are similar to the apparent W(m) values determined using the Guggenheim-Anderson-DeBoer (GAB) equation (0.110, 0.117, and 0.147 weight fraction of water for PVP, PVP-PEG 10%, and PVP-PEG 20%, respectively). Nonfreezing water is detected above 0.47, 0.49, and 0.51 weight fraction of water for PVP, PVP-PEG 10%, and PVP-PEG 20%, respectively. Miscibility between water and PVP or PVP-PEG seems to change according to the water content in the system. All parameters increase with the concentration of PEG in the sample. This may be explained by the fact that PEG has a larger number of polymer repeating units, which may therefore interact with water more than PVP.


Stress and morale in general practice: a comparison of two health care systems.
Poor morale and high levels of stress among general practitioners (GPs) are causing concern. Little research has previously been carried out to study possible differences in morale and stress between GPs working in two different but geographically similar health care systems. To compare perceived levels of stress and morale between GPs working in two different health care systems--one having a state monopoly (Northern Ireland) and the other having mixed private and state funding (Republic of Ireland)--and to look for factors that might help explain any differences in stress levels and morale between the two systems. Anonymous and confidential questionnaires were sent to all 986 National Health Service (NHS) GPs in Northern Ireland (NI) and a random sample of 900 GPs in the Republic of Ireland (ROI). A common set of core questions on demographic details, partners and work patterns, perceived levels of stress and morale, safety, violence, and complaints were asked. Response rates were high in both areas: 91% in NI and 78% in the ROI. GPs in NI had significantly higher stress levels and significantly lower levels of morale than GPs in the ROI. The NI sample expect matters to get worse over the following year. Doctors in the ROI were more likely to be single handed and to work from two sites. Also, more GPs in ROI had fears for their safety and had been the subject of physical violence, but fewer had received complaints and medico-legal actions than in NI. A significant proportion of both groups of doctors report feeling highly stressed but GPs in NI appear more unhappy and have a poorer outlook for the future. It is suggested that the structure, management, and expectations of the NHS have taken a severe toll on its GPs, whereas a system in which doctors have less practice support but more control is good for morale.


Standing genetic variation drives repeatable experimental evolution in outcrossing populations of Saccharomyces cerevisiae.
In "evolve-and-resequence" (E&R) experiments, whole-genome sequence data from laboratory-evolved populations can potentially uncover mechanisms of adaptive change. E&R experiments with initially isogenic, asexually reproducing microbes have repeatedly shown that beneficial de novo mutations drive adaptation, and these mutations are not shared among independently evolving replicate populations. Recent E&R experiments with higher eukaryotes that maintain genetic variation via sexual reproduction implicate largely different mechanisms; adaptation may act primarily on pre-existing genetic variation and occur in parallel among independent populations. But this is currently a debated topic, and generalizing these conclusions is problematic because E&R experiments with sexual species are difficult to implement and important elements of experimental design suffer for practical reasons. We circumvent potentially confounding limitations with a yeast model capable of shuffling genotypes via sexual recombination. Our starting population consisted of a highly intercrossed diploid Saccharomyces cerevisiae initiated from four wild haplotypes. We imposed a laboratory domestication treatment on 12 independent replicate populations for 18 weeks, where each week included 2 days as diploids in liquid culture and a forced recombination/mating event. We then sequenced pooled population samples at weeks 0, 6, 12, and 18. We show that adaptation is highly parallel among replicate populations, and can be localized to a modest number of genomic regions. We also demonstrate that despite hundreds of generations of evolution and large effective population sizes, de novo beneficial mutations do not play a large role in this adaptation. Further, we have high power to detect the signal of change in these populations but show how this power is dramatically reduced when fewer timepoints are sampled, or fewer replicate populations are analyzed. As ours is the most highly replicated and sampled E&R study in a sexual species to date, this evokes important considerations for past and future experiments.


Derivatives of salicylic acid as inhibitors of YopH in Yersinia pestis.
Yersinia pestis causes diseases ranging from gastrointestinal syndromes to bubonic plague and could be misused as a biological weapon. As its protein tyrosine phosphatase YopH has already been demonstrated as a potential drug target, we have developed two series of forty salicylic acid derivatives and found sixteen to have micromolar inhibitory activity. We designed these ligands to have two chemical moieties connected by a flexible hydrocarbon linker to target two pockets in the active site of the protein to achieve binding affinity and selectivity. One moiety possessed the salicylic acid core intending to target the phosphotyrosine-binding pocket. The other moiety contained different chemical fragments meant to target a nearby secondary pocket. The two series of compounds differed by having hydrocarbon linkers with different lengths. Before experimental co-crystal structures are available, we have performed molecular docking to predict how these compounds might bind to the protein and to generate structural models for performing binding affinity calculation to aid future optimization of these series of compounds.


Amelioration of alcohol-induced hepatotoxicity and oxidative stress in rats by Acorus calamus.
The protective effect of a methanolic extract (ME) of Acorus calamus against alcohol-induced hepatotoxicity and oxidative stress was studied in rats. The in vitro assays using DPPH and ABTS showed a strong antioxidant activity of the extract with the total polyphenolic content of 156 mg/g. Chronic ethanol administration causes an increase in oxidative stress and tissue injury with decreased antioxidant status. In this study, continuous administration of ethanol (7.9 g/kg body weight/day) for a period of 6 weeks resulted in a significant (p < .001) increase in the levels of serum aspartate aminotransferase, serum alanine aminotransferase, alkaline phospahatase, and bilirubin with the decreased level of total antioxidant status. Moreover, the levels of lipid peroxidation markers (malondialdehyde and hydroperoxides) as well as protein carbonyl content were also increased (p < .001), whereas the levels of non-enzymic antioxidants (glutathione, vitamin C, and vitamin E) decreased significantly in the liver tissues of ethanol-administered control rats. Pretreatment of rats with ME at doses of 300 and 600 g/kg body weight before alcohol administration significantly reduced the hepatic marker enzymes, level of lipid peroxidation, and protein oxidation, and increased the enzymatic and non-enzymatic antioxidant levels in liver. These observations were supplemented by histopathological examination of liver sections. Overall, the present study shows that the administration of ME ameliorates the antioxidant status as well as protects against the toxic effects of ethanol in rats, thereby suggesting its use as an effective botanical supplement for hepatoprotection.