Novel concepts in the development of platinum antitumor drugs.
The limitations of cisplatin as an anticancer drug have stimulated the search for other antitumor-active platinum complexes with improved pharmacological properties. The two main goals in the search for new platinum anti-cancer agents are the reduction of the dose-limiting toxicities of cisplatin and the circumvention of cisplatin resistance. However, it should be pointed out that this has proven to be a difficult task. In fact, less than 1% of the thousand of platinum complexes tested for pre-clinical antitumor activity have entered clinical trials in the past 30 years. Nonetheless, right now, several new platinum complexes are in clinical trials, a proof of the continued belief that platinum complexes may still fulfil the needs for novel antitumor drugs. This review will focus on the three main innovative approaches found in the platinum anticancer-field, namely, (1) compounds with decreased reactivity against nucleophiles, (2) compounds with carrier ligands, and (3) compounds which bind differently to DNA as compared to cisplatin. In the latter class, special attention is paid to dinuclear and polinuclear platinum complexes.


Effects of alpha- and beta-adrenergic agonists on intramammary pressure and milk flow in dairy cows.
alpha-Adrenergic mimetics inhibit milk removal whereas beta-adrenergic mimetics have a favourable effect upon milk removal in dairy cows. This work aimed to relate these effects to the milk ejection reflex mechanism and/or milk flow from the cisternal cavities only. The alpha-adrenergic agonist phenylephrine and the beta-adrenergic agonist isoproterenol were injected prior to and after milk ejection. Applied after milk ejection, the phenylephrine treatment reduced intramammary pressure (IMP) and milk yield markedly. Phenylephrine application before milk ejection prevented IMP from rising during teat stimulation or after oxytocin injection. Milking produced small amounts of milk at minimal milk flow rates. The very different responses to phenylephrine application prior to and after milk ejection indicate separate adrenergic inhibitory mechanisms for milk ejection and milk flow. The application of the beta-adrenergic agonist isoproterenol did not alter the IMP. Nevertheless, this compound improved milk removal by increasing milk flow. The observation of bimodal milk flow at milkings without preceding teat stimulation indicates a delayed milk ejection. Isoproterenol injection without teat stimulation enhanced milk flow rates, but the bimodality remained unchanged. We conclude that isoproterenol enhances milk flow without affecting milk ejection.


Broadband optical parametric amplification at the communication band with periodically poled lithium niobate.
We report broadband optical parametric generation (OPG) in a single periodically poled lithium niobate crystal with a picosecond pump pulse at a fixed wavelength. We also demonstrate efficient optical parametric amplification of a broadband seed pulse within the quasi-phase-matched OPG band. The broad parametric gain band is attributed to group-velocity matching and degeneracy between the signal and idler, and the broad spectral width of the pumping source.


Fabrication of Au-Nanoparticle-Embedded Lipid Bilayer Membranes Supported on Solid Substrates.
We fabricated gold nanoparticle (Au-NP)-embedded supported lipid bilayers (SLBs) by two methods. In the vesicle-vesicle fusion method, vesicles with hydrophobized Au-NPs are ruptured and fused on SiO2/Si substrates. In the vesicle-membrane fusion method, SLBs without Au-NPs were preformed on the substrate and then vesicles with Au-NPs were fused into the preformed membranes. In the former method, Au-NP incorporation into the SLBs was observed as an increase in the membrane thickness in atomic force microscopy (AFM) images and directly observed by transmission electron microscopy. In the latter method, fusion of vesicles into the preformed membranes was confirmed by the fluorescent color change in the preformed membranes, and Au-NP incorporation was also confirmed by an increase in the membrane thickness in the AFM images. Key techniques for the successful vesicle-membrane fusion are hydrophobization of Au-NPs, approach control of vesicles by mixing the charged lipids, and destabilization of the lipid bilayers by adding lipids with a small polar headgroup.


Disproportionation of Inorganic Sulfur Compounds by Mesophilic Chemolithoautotrophic Campylobacterota.
The disproportionation of inorganic sulfur compounds could be widespread in natural habitats, and microorganisms could produce energy to support primary productivity through this catabolism. However, the microorganisms that carry this process out and the catabolic pathways at work remain relatively unstudied. Here, we investigated the bacterial diversity involved in sulfur disproportionation in hydrothermal plumes from Carlsberg Ridge in the northwestern Indian Ocean by enrichment cultures. A bacterial community analysis revealed that bacteria of the genera Sulfurimonas and Sulfurovum, belonging to the phylum Campylobacterota and previously having been characterized as chemolithoautotrophic sulfur oxidizers, were the most dominant members in six enrichment cultures. Subsequent bacterial isolation and physiological studies confirmed that five Sulfurimonas and Sulfurovum isolates could disproportionate thiosulfate and elemental sulfur. The ability to disproportionate sulfur was also demonstrated in several strains of Sulfurimonas and Sulfurovum that were isolated from hydrothermal vents or other natural environments. Dialysis membrane experiments showed that S0 disproportionation did not require the direct contact of cells with bulk sulfur. A comparative genomic analysis showed that Campylobacterota strains did not contain some genes of the Dsr and rDSR pathways (aprAB, dsrAB, dsrC, dsrMKJOP, and qmoABC) that are involved in sulfur disproportionation in some other taxa, suggesting the existence of an unrevealed catabolic pathway for sulfur disproportionation. These findings provide evidence for the catabolic versatility of these Campylobacterota genera, which are widely distributed in chemosynthetic environments, and expand our knowledge of the microbial taxa involved in this reaction of the biogeochemical sulfur cycle in hydrothermal vent environments. IMPORTANCE The phylum Campylobacterota, notably represented by the genera Sulfurimonas and Sulfurovum, is ubiquitous and predominant in deep-sea hydrothermal systems. It is well-known to be the major chemolithoautotrophic sulfur-oxidizing group in these habitats. Herein, we show that the mesophilic predominant chemolithoautotrophs of the genera Sulfurimonas and Sulfurovum could grow via sulfur disproportionation to gain energy. This is the first report of the chemolithoautotrophic disproportionation of thiosulfate and elemental sulfur within the genera Sulfurimonas and Sulfurovum, and this comes in addition to their already known role in the chemolithoautotrophic oxidation of sulfur compounds. Sulfur disproportionation via chemolithoautotrophic Campylobacterota may represent a previously unrecognized primary production process in hydrothermal vent ecosystems.


Worldwide variation in the doubling time of Alzheimer's disease incidence rates.
The doubling time is the number of chronological years for the age-specific incidence rate to double in magnitude. Doubling times describe the rate of increase of the risk of Alzheimer's disease (AD) with advancing age. Estimates of doubling times of AD assist in understanding disease etiology and forecasting future disease prevalence. The objective of this study was to investigate regional and gender differences in the doubling of AD age-specific incidence rates. We identified all studies in the peer review literature that reported age-specific incidence rates for AD. We modeled the logarithm of the incidence rate as a linear function of age. We used both fixed effects models and random effects models to account for interstudy variation. AD incidence rates exponentially increase with increasing age. The overall estimate of the doubling time was 5.5 years. The doubling times from studies performed in North America and Europe were 6.0 and 5.8, respectively; whereas the doubling times in all other parts of the world were 5.0. There was no significant geographic differences in doubling times (P = .3). Although the doubling times were slightly longer for men (6.5 years) than for women (5.4 years), the difference was not significant (P = .3). Doubling times of AD incidence rates are not statistically significantly different among populations throughout the world. The risk of AD grows exponentially with age, doubling approximately every 5 to 6 years. Although the shapes of the incidence curves are similar, there is considerable variation in absolute incidence rates throughout the world. Currently, there are limited epidemiologic data at the oldest ages, and further study is needed to accurately define the incidence curve above age 90.


Cardiovascular events during differing hypertension therapies in patients with diabetes.
The aim of this study was to determine which combination therapy in patients with hypertension and diabetes most effectively decreases cardiovascular events. The ACCOMPLISH (Avoiding Cardiovascular Events Through COMbination Therapy in Patients Living With Systolic Hypertension) trial compared the outcomes effects of a renin-angiotensin system blocker, benazepril, combined with amlodipine (B+A) or hydrochlorothiazide (B+H). A separate analysis in diabetic patients was pre-specified. A total of 6,946 patients with diabetes were randomized to treatment with B+A or B+H. A subgroup of 2,842 diabetic patients at very high risk (previous cardiovascular or stroke events) was also analyzed, as were 4,559 patients without diabetes. The primary end point was a composite of cardiovascular death, myocardial infarction, stroke, hospitalization for angina, resuscitated arrest, and coronary revascularization. In the full diabetes group, the mean achieved blood pressures in the B+A and B+H groups were 131.5/72.6 and 132.7/73.7 mm Hg; during 30 months, there were 307 (8.8%) and 383 (11.0%) primary events (hazard ratio [HR]: 0.79, 95% confidence interval [CI]: 0.68 to 0.92, p = 0.003). For the diabetic patients at very high risk, there were 195 (13.6%) and 244 (17.3%) primary events (HR: 0.77, 95% CI: 0.64 to 0.93, p = 0.007). In the nondiabetic patients, there were 245 (10.8%) and 296 (12.9%) primary events (HR: 0.82, 95% CI: 0.69 to 0.97, p = 0.020). In the diabetic patients, there were clear coronary benefits with B+A, including both acute clinical events (p = 0.013) and revascularizations (p = 0.024). There were no unexpected adverse events. In patients with diabetes and hypertension, combining a renin-angiotensin system blocker with amlodipine, compared with hydrochlorothiazide, was superior in reducing cardiovascular events and could influence future management of hypertension in patients with diabetes. (Avoiding Cardiovascular Events Through COMbination Therapy in Patients Living With Systolic Hypertension [ACCOMPLISH]; NCT00170950).


Clinical efficacy of micronized purified flavonoid fraction: an overview.
Flavonoids are widely used for the treatment of chronic venous insufficiency (CVI). In clinical trials, micronized purified flavonoid fraction (MPFF, 450 mg diosmin plus 50 mg hesperidin, Daflon 500 mg) has demonstrated its activity in CVI by improving venous tone and vein elasticity assessed by plethysmography. Randomized, double-blind, placebo-controlled clinical studies have shown an improvement in signs and symptoms related to CVI and a decrease in leg circumferences at the levels of the ankle and calf. The effect of MPFF on microcirculatory parameters suspected of participating in the pathophysiological process of venous ulceration has been investigated in patients. These include hemorheological parameters and transcutaneous oxygen tension measurements, which were shown to improve after treatment with MPFF. Finally, a randomized double-blind, placebo-controlled clinical study has shown that MPFF treatment for 2 months, in addition to standard compression therapy, accelerates leg ulcer healing in patients with ulcers </=10 cm.


Comparative glycomic mapping through quantitative permethylation and stable-isotope labeling.
Comparative glycan quantification has thus far been a challenging task due to the lack of sensitive and reproducible analytical techniques. We introduce here a combination of quantitative permethylation and isotope labeling of glycans as an approach (C-GlycoMAP) allowing precise comparison between different samples in a single MALDI-MS analysis. Samples are either methylated or deuteriomethylated prior to their mixing and mass spectrometric acquisitions. Comparative analyses are based on the ratio of the two isotopically distinct forms of the same glycan structure, thus allowing a direct absolute evaluation of the intensities of the two forms originating from two different biological samples (e.g., control and diseased). The direct comparison between the two forms eliminates a MALDI-MS low m/z bias commonly associated with this technique. These comparative analyses are highly reliable when the intensity ratios of the two forms lie between 0.125 and 6, an overall reproducibility better than 30% (RSD). The value of C-GlycoMAP is demonstrated here for N-glycans derived from human blood serum collected from a healthy individual and a breast cancer patient as well as for O-glycans derived from normal and cancer cell extracts.


Anti-Sa antibodies: prognostic and pathogenetic significance to rheumatoid arthritis.
Anti-Sa antibodies are detected in the serum of 20-47% of patients with rheumatoid arthritis. These antibodies have a high degree of specificity for the disease, and appear to identify a subset of early rheumatoid arthritis patients destined to have aggressive and destructive disease. It has recently been confirmed that anti-Sa antibodies are directed to citrullinated vimentin, thus placing them in the anti-citrulline family of autoantibodies. The Sa antigen has previously been shown to be present in synovium. This, along with the demonstration of citrullinated proteins in rheumatoid synovium, suggests that anti-Sa antibodies may play a pathogenetic role in the initiation and/or persistence of rheumatoid synovitis.


Position of the Academy of Nutrition and Dietetics: Obesity, Reproduction, and Pregnancy Outcomes.
It is the position of the Academy of Nutrition and Dietetics that all women of reproductive age receive education about maternal and fetal risks associated with prepregnancy obesity, excessive gestational weight gain, and significant postpartum weight retention, including potential benefits of lifestyle changes. Behavioral counseling to improve dietary intake and physical activity should be provided to overweight and obese women, beginning in the preconception period and continuing throughout pregnancy, for at least 12 to 18 months postpartum. Weight loss before pregnancy may improve fertility and reduce the risk of poor maternal-fetal outcomes, such as preterm birth, gestational diabetes, gestational hypertension, pre-eclampsia, assisted delivery, and select congenital anomalies. Lifestyle interventions that moderate gestational weight gain may reduce the risk of poor pregnancy outcomes, such as gestational diabetes, gestational hypertension, large for gestational age, and macrosomia, as well as lower the risk for significant postpartum retention. Postpartum interventions that promote healthy diet and physical activity behaviors may reduce postpartum weight retention and decrease obesity-related risks in subsequent pregnancies. Analysis of the evidence suggests that there is good evidence to support the role of diet, physical activity, and behavior changes in promoting optimal weight gain during pregnancy; however, there is currently a relative lack of evidence in other areas related to reproductive outcomes.


Optically monitored dip coating as a contactless viscometry method for liquid films.
Real-time interferometric monitoring of the dip coating process is applied to the study of properties of flowing liquids. Nonvolatile Newtonian oils are considered, allowing validity of a simple model after the steady state is reached where film physical thickness depends on time as t(-1/2). Measurement of two distinct mineral oil standards, under several withdrawing speeds, resulted in kinematic viscosities of 1.17+/-0.03 and 9.9+/-0.2 S (1S = 1 cm2/s). Agreement of these results with nominal values from the manufacturer suggests that interferometric monitoring of dip coating may become a valuable method for accurate, contactless viscometry of liquid films. Advantages and present limitations are discussed.


Risk of pancreatitis with mutation of the cystic fibrosis gene.
Between 5% and 15% of patients with recurrent pancreatitis have no identified etiology after routine investigation and advanced endoscopic evaluation. To determine whether mutation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene is a risk factor for idiopathic pancreatitis. We compared the frequency of CFTR mutations as measured by DNA probe analysis in a case group of persons with idiopathic pancreatitis and a control group without pancreatitis, all of whom underwent endoscopic retrograde cholangiopancreatography. A separate analysis compared the prevalence of CFTR mutations between the case group and controls with pancreatitis of known etiology. A subgroup comparison was made between cases of pancreas divisum with pancreatitis and controls with pancreas divisum and no pancreatitis. CFTR mutations were present in 19 (19%) of 96 cases and 7 (3.5%) of 198 controls without pancreatitis (odds ratio, OR = 6.7; 95% CI, 2.8-16.3; p < 0.00001). Compared to the controls with a known cause of pancreatitis (N = 78), cases had a higher prevalence of CFTR mutations (19% vs 2.6%, OR = 9.4; CI, 2.1-41.7; p= 0.0005). Among subjects with pancreas divisum, CFTR mutations were present in 8 (22%) of 37 cases compared to 0 (0%) of 20 controls (OR = 11.8; CI, 8.9-14.7; p= 0.02). The risk of idiopathic pancreatitis is greater among persons with CFTR mutations as compared to persons without CFTR mutations. Among persons with pancreas divisum, CFTR mutations appear to increase the risk for pancreatitis.


[Prosthodontics technological processes and results of risk profiling].
In prosthodontics laboratories, dental devices and appliances are made to measure, optimized and tested, in particular prostheses, following the design specifications of the dentist. He purpose of this study was to learn about and become familiar with the work processes in prosthodontics laboratories and also analyze inconveniences and risks in the laboratory work space. The simple computerized pre-mapping model, the risk profiling method, was used. Repetitive movement evaluation was further analyzed using the multitask OCRA mini-checklist. Repeated movement risk to the upper limbs was identified during some production processes; chemical and physical pollutants were present as well as problems related to the use of certain equipment. This by no means exhaustive analysis can be considered as a starting point in creating guidelines in risk evaluation in a sector consisting of small and very small businesses which has so far not been studied in depth.


Optimizing Cold Water Immersion for Exercise-Induced Hyperthermia: A Meta-analysis.
Cold water immersion (CWI) provides rapid cooling in events of exertional heat stroke. Optimal procedures for CWI in the field are not well established. This meta-analysis aimed to provide structured analysis of the effectiveness of CWI on the cooling rate in healthy adults subjected to exercise-induced hyperthermia. An electronic search (December 2014) was conducted using the PubMed and Web of Science. The mean difference of the cooling rate between CWI and passive recovery was calculated. Pooled analyses were based on a random-effects model. Sources of heterogeneity were identified through a mixed-effects model Q statistic. Inferential statistics aggregated the CWI cooling rate for extrapolation. Nineteen studies qualified for inclusion. Results demonstrate CWI elicited a significant effect: mean difference, 0.03°C·min(-1); 95% confidence interval, 0.03-0.04°C·min(-1). A conservative, observed estimate of the CWI cooling rate was 0.08°C·min(-1) across various conditions. CWI cooled individuals twice as fast as passive recovery. Subgroup analyses revealed that cooling was more effective (Q test P < 0.10) when preimmersion core temperature ≥38.6°C, immersion water temperature ≤10°C, ambient temperature ≥20°C, immersion duration ≤10 min, and using torso plus limbs immersion. There is insufficient evidence of effect using forearms/hands CWI for rapid cooling: mean difference, 0.01°C·min(-1); 95% confidence interval, -0.01°C·min(-1) to 0.04°C·min(-1). A combined data summary, pertaining to 607 subjects from 29 relevant studies, was presented for referencing the weighted cooling rate and recovery time, aiming for practitioners to better plan emergency procedures. An optimal procedure for yielding high cooling rates is proposed. Using prompt vigorous CWI should be encouraged for treating exercise-induced hyperthermia whenever possible, using cold water temperature (approximately 10°C) and maximizing body surface contact (whole-body immersion).


Autophagy is activated in injured neurons and inhibited by methylprednisolone after experimental spinal cord injury.
Experimental, controlled trial, animal study. To assess autophagy expression after rat spinal cord injury (SCI) and investigate the effect of methylprednisolone treatment on autophagy. Although it is evident that SCI induces necrosis and apoptosis, its relationship to autophagy is uncertain. Autophagy is implicated in various pathological states in the nervous system, such as neurodegenerative diseases, cerebral ischemia, and traumatic brain injury. Up to now, no autophagy expression was evidenced by transmission electronic microscope (TEM) and the autophagy marker, microtubule-associated protein light chain 3 (LC3) in neural tissue after SCI. Sixty-six Sprague-Dawley rats were used for the experimental procedure. In the SCI group, laminectomy at T9 were performed, followed by impactor contusion of the spinal cord. In the sham group, only a laminectomy was performed without contusion. We used Western blot to analyze LC3 at 2 hours, 4 hours, 1 day, 3 days, and 7 days after SCI. We also investigated the effect of methylprednisolone on autophagy expression of contused spinal cord. Cellular localization and ultrastructural changes after spinal cord injury were compared with those sham-operated rats using immunofluorescent double labeling and TEM, respectively. Data from the Western blot were analyzed using a nonparametric Kruskal-Wallis test with P < 0.05 being considered significant. We detected significantly elevated level of LC3 2 hours after SCI, and then the level declined until 1 week after SCI. Methylprednisolone decreased LC3 expression at 2 hours after SCI. LC3 positive cells were colocalized with neuronal nuclei, but not with glial fibrillary acidic protein. The existence of autophagy and progress of autophagic cell death after SCI were confirmed by TEM. Through observing the enhanced autophagy expression in neurons soon after contusion injury and the inhibitive effect of methylprednisolone treatment, this study demonstrates the characteristics of autophagy expression after SCI and suggests that autophagic cell death may play a role in neuronal death after spinal cord trauma.


A Strategy to Replace the Mouse Bioassay for Detecting and Identifying Lipophilic Marine Biotoxins by Combining the Neuro-2a Bioassay and LC-MS/MS Analysis.
Marine biotoxins in fish and shellfish can cause several symptoms in consumers, such as diarrhea, amnesia, or even death by paralysis. Monitoring programs are in place for testing shellfish on a regular basis. In some countries testing is performed using the so-called mouse bioassay, an assay that faces ethical concerns not only because of animal distress, but also because it lacks specificity and results in high amounts of false positives. In Europe, for lipophilic marine biotoxins (LMBs), a chemical analytical method using LC-MS/MS was developed as an alternative and is now the reference method. However, safety is often questioned when relying solely on such a method, and as a result, the mouse bioassay might still be used. In this study the use of a cell-based assay for screening, i.e., the neuro-2a assay, in combination with the official LC-MS/MS method was investigated as a new alternative strategy for the detection and quantification of LMBs. To this end, samples that had been tested previously with the mouse bioassay were analyzed in the neuro-2a bioassay and the LC-MS/MS method. The neuro-2a bioassay was able to detect all LMBs at the regulatory levels and all samples that tested positive in the mouse bioassay were also suspect in the neuro-2a bioassay. In most cases, these samples contained toxin levels (yessotoxins) that explain the outcome of the bioassay but did not exceed the established maximum permitted levels.


Inhibitory effects of citronellol and geraniol on nitric oxide and prostaglandin E₂production in macrophages.
Geranium oil has been used traditionally for diarrhea, dermatitis, and intestinal inflammation in East Asia. The aim of this study was to determine the effects of geranium oil's characteristic components, citronellol and geraniol, on lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in RAW 264.7 macrophages. Citronellol and geraniol suppressed NO and PGE(2) production in a dose-dependent manner. The inhibitory efficacy of geraniol was concomitant with decreases in protein and mRNA expression levels of inducible nitric oxide synthase (iNOS), whereas citronellol inhibited only iNOS enzymatic activity. By adding citronellol and geraniol, the LPS-induced cyclooxygenase-2 (COX-2) protein and mRNA expression levels were significantly attenuated, whereas cytosolic degradation of I κB α and upregulation of NF-κB p65 in the nucleus were reversed. These results suggested that citronellol and geraniol exhibit anti-inflammatory activities, supporting their common use and demonstrating their therapeutic potential for inflammation-associated disorders.


The effects of dynamic exercise utilizing PNF patterns on abdominal muscle thickness in healthy adults.
[Purpose] The purpose of this study is to examine the effects of dynamic exercise utilizing the PNF (proprioceptor neuromuscular facilitation) patterns accompanied by abdominal drawing-in exercises on abdominal muscle thickness in healthy adults. [Subjects] The total number of subjects was 30;15 were randomly placed in the training group (TG), and the remaining 15 made up the control group (CG). [Methods] The subjects in the TG conducted 3-5 sets of dynamic exercises utilizing the PNF patterns each day, 3 times a week for 6 weeks. The thickness of the abdominal muscles was measured by ultrasonography. [Results] When the TG's abdominal muscle thickness pre-test and post-test were compared in this study, there was a statistical significance in all of the external obliquus abdominis (Eo), the internal obliquus abdominis (Io), and the transversus abdominis (Tra). [Conclusion] Dynamic exercise utilizing the PNF patterns increased the thickness of the abdominal muscles that are the basis of trunk stabilization.


Healing potential of zerumbone ointment on experimental full-thickness excision cutaneous wounds in rat.
Zerumbone is a monocyclic sesquiterpene compound. Based on report, it is the predominant bioactive compound from the rhizomes of Zingiber zerumbet. The study was undertaken to evaluate the therapeutic effects of topical zerumbone on excision wounds in rats. A 1% (w/w) simple ointment containing zerumbone was applied topically (100 mg ointment per rat) once a day on full-thickness excision wounds created on rats. The wound tissue was removed and used for estimation of antioxidant activity and to observe histopathological changes. Immunohistochemical staining was performed to study the expression pattern of vascular endothelial growth factor (VEGF), transforming growth factor (TGF)-β1 and collagen IV. Zerumbone exhibited antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Candida albicans and Candida tropicali. Zerumbone ointment has potent wound healing capacity as evident from the wound contraction on 15th post wounding day. The histopathological examinations of healed wound sections showed greater tissue regeneration, more fibroblasts and angiogenesis in zerumbone ointment-treated group. VEGF, TGF-β1 and collagen IV expression were also correlative with the healing pattern observed. Zerumbone possesses potent antioxidant activity by increasing superoxide dismutase, catalase, glutathione and decreased lipid peroxidation. The synergistic effects of both antimicrobial and antioxidant activities in zerumbone are deduced to have accelerated the wound repair. The results demonstrate that zerumbone possessed strong wound healing potential and can be exploited to accelerate excision wound healing.


[Non-histone skeleton of mitotic chromosome in situ].
Studying giant nuclei of Chironomus plumosus in situ (Makarov, Chentsov, 2010), we concluded that polythene chromosome structure appears after 2 M NaCl and DNase treatment in presence of 2 mM CuCl2. Cu2+ -ions may stabilize bonds between specific non-histone components, arranged into non-histone matrix of polythene chromosome. Here, we investigated the non-histone matrix of pig embryo mitotic chromosomes in situ, using 2 mM CuCl2-stabilization method. In 2 mM CuCl2-stabilized cells the residual chromosome body (non-histone matrix) could be visualized in every stage of mitosis. Mitotic chromosome non-histone matrix had the same reaction on preliminary hypotonic treatment as normal chromosome: different decondensation of non-histone material was observed. Topoisomerase IIalpha and SMC 1 had uniform localization inside chromosomal body and did not form any axial structures.


Prediction of the coding sequences of unidentified human genes. IV. The coding sequences of 40 new genes (KIAA0121-KIAA0160) deduced by analysis of cDNA clones from human cell line KG-1.
In this series of projects regarding the accumulation of sequence information of unidentified human genes, we newly deduced the sequences of 40 full-length cDNA clones of human cell line KG-1, and predicted the coding sequences of the corresponding genes, named KIAA0121 to 0160. The results of a computer search of public databases indicated that the sequences of 13 genes were unrelated to any reported genes, while the remaining 27 genes carried sequences which showed some similarities to known genes. Obvious unique sequences noted were as follows. A stretch of triplet repeats was contained in each of three genes: These were GAG(Glu) in KIAA0122 and KIAA0147, and TCC(Ser) in KIAA0150. A stretch of 10 amino acid-residues was repeated 21 times in KIAA0139, and a homologous sequence of 76-78 nucleotides was found repeated 6 times in the untranslated region of KIAA0125. Northern hybridization analysis demonstrated that 13 genes were expressed in a cell- or tissue-specific manner. Although a vast number of expressed sequence tags (ESTs) have been registered for comprehensive analysis of cDNA clones, our sequence data indicated that their distribution is very unbalanced: e.g. while no EST hit 7 genes, 85 ESTs fell in a single gene.


Serum stem cell factor level in renal transplant recipients with posttransplant erythrocytosis.
The etiology of posttransplant erythrocytosis (PTE) remains unclear, and the most frequently suggested causative factors are still a matter of controversy. We aimed to investigate serum-soluble stem cell factor (sSCF) along with serum erythropoietin (EPO) levels in renal transplant recipients (RTRs) with PTE. Thirteen RTRs with PTE, 42 RTRs without PTE, and 42 healthy controls were included. Serum sSCF and EPO levels were determined using an enzyme-linked immunosorbent assay kit. Expected and observed/expected EPO levels were calculated. Serum sSCF levels and observed/expected EPO were significantly higher in RTRs with PTE than both RTRs without PTE and controls. In RTRs with PTE, sSCF level was significantly correlated with hematocrit and observed/expected EPO, respectively. Significant correlation was also observed between hematocrit level and observed/expected EPO in RTRs with PTE. Increased sSCF level and inadequate suppression of EPO production seem to have a role in the pathogenesis of PTE.


Secondary reinforcement and number of primary reinforcements.
Pigeons' pecks on either of two concurrently available response keys produced secondary reinforcers according to independent one-minute variable-interval schedules. Different secondary reinforcers, in the presence of which the rates of primary reinforcement were equal, were associated with each key. The rate of pecking maintained by each secondary reinforcer varied directly, but nonproportionally, with the number of primary reinforcements given in the presence of the secondary reinforcer.


Highly efficient site-specific transgenesis in cancer cell lines.
Transgenes introduced into cancer cell lines serve as powerful tools for identification of genes involved in cancer. However, the random nature of genomic integration site of a transgene highly influences the fidelity, reliability and level of its expression. In order to alleviate this bottleneck, we characterized the potential utility of a novel PhiC31 integrase-mediated site-specific insertion system (PhiC31-IMSI) for introduction of transgenes into a pre-inserted docking site in the genome of cancer cells. According to this system, a "docking-site" was first randomly inserted into human cancer cell lines and clones with a single copy were selected. Subsequently, an "incoming" vector containing the gene of interest was specifically inserted in the docking-site using PhiC31. Using the Pc-3 and SKOV-3 cancer cell lines, we showed that transgene insertion is reproducible and reliable. Furthermore, the selection system ensured that all surviving stable transgenic lines harbored the correct integration site. We demonstrated that the expression levels of reporter genes, such as green fluorescent protein and luciferase, from the same locus were comparable among sister, isogenic clones. Using in vivo xenograft studies, we showed that the genetically altered cancer cell lines retain the properties of the parental line. To achieve temporal control of transgene expression, we coupled our insertion strategy with the doxycycline inducible system and demonstrated tight regulation of the expression of the antiangiogenic molecule sFlt-1-Fc in Pc-3 cells. Furthermore, we introduced the luciferase gene into the insertion cassette allowing for possible live imaging of cancer cells in transplantation assays. We also generated a series of Gateway cloning-compatible intermediate cassettes ready for high-throughput cloning of transgenes and demonstrated that PhiC31-IMSI can be achieved in a high throughput 96-well plate format. The novel PhiC31-IMSI system described in this study represents a powerful tool that can facilitate the characterization of cancer-related genes.


Correlation of Impedance Threshold Device use during cardiopulmonary resuscitation with post-cardiac arrest Acute Kidney Injury.
To assess whether use of Impedance Threshold Device (ITD) during cardiopulmonary resuscitation (CPR) reduces the degree of post-cardiac arrest Acute Kidney Injury (AKI), as a result of improved hemodynamics, in a porcine model of ventricular fibrillation (VF) cardiac arrest. After 8 min of untreated cardiac arrest, the animals were resuscitated either with active compression-decompression (ACD) CPR plus a sham ITD (control group, n=8) or with ACD-CPR plus an active ITD (ITD group, n=8). Adrenaline was administered every 4 min and electrical defibrillation was attempted every 2 min until return of spontaneous circulation (ROSC) or asystole. After ROSC the animals were monitored for 6 h under general anesthesia and then returned to their cages for a 48 h observation, before euthanasia. Two novel biomarkers, Neutrophil Gelatinase-Associated Lipocalin (NGAL) in plasma and Interleukin-18 (IL-18) in urine, were measured at 2 h, 4 h, 6 h, 24 h and 48 h post-ROSC, in order to assess the degree of AKI. ROSC was observed in 7 (87.5%) animals treated with the sham valve and 8 (100%) animals treated with the active valve (P=NS). However, more than twice as many animals survived at 48 h in the ITD group (n=8, 100%) compared to the control group (n=3, 37.5%). Urine IL-18 and plasma NGAL levels were augmented post-ROSC in both groups, but they were significantly higher in the control group compared with the ITD group, at all measured time points. Use of ITD during ACD-CPR improved hemodynamic parameters, increased 48 h survival and decreased the degree of post-cardiac arrest AKI in the resuscitated animals.


Listeria monocytogenes internalin B activates junctional endocytosis to accelerate intestinal invasion.
Listeria monocytogenes (Lm) uses InlA to invade the tips of the intestinal villi, a location at which cell extrusion generates a transient defect in epithelial polarity that exposes the receptor for InlA, E-cadherin, on the cell surface. As the dying cell is removed from the epithelium, the surrounding cells reorganize to form a multicellular junction (MCJ) that Lm exploits to find its basolateral receptor and invade. By examining individual infected villi using 3D-confocal imaging, we uncovered a novel role for the second major invasin, InlB, during invasion of the intestine. We infected mice intragastrically with isogenic strains of Lm that express or lack InlB and that have a modified InlA capable of binding murine E-cadherin and found that Lm lacking InlB invade the same number of villi but have decreased numbers of bacteria within each infected villus tip. We studied the mechanism of InlB action at the MCJs of polarized MDCK monolayers and find that InlB does not act as an adhesin, but instead accelerates bacterial internalization after attachment. InlB locally activates its receptor, c-Met, and increases endocytosis of junctional components, including E-cadherin. We show that MCJs are naturally more endocytic than other sites of the apical membrane, that endocytosis and Lm invasion of MCJs depends on functional dynamin, and that c-Met activation by soluble InlB or hepatocyte growth factor (HGF) increases MCJ endocytosis. Also, in vivo, InlB applied through the intestinal lumen increases endocytosis at the villus tips. Our findings demonstrate a two-step mechanism of synergy between Lm's invasins: InlA provides the specificity of Lm adhesion to MCJs at the villus tips and InlB locally activates c-Met to accelerate junctional endocytosis and bacterial invasion of the intestine.


Aminorex and pulmonary hypertension. A review.
There was an epidemic of chronic pulmonary hypertension in Austria, the Federal Republic of Germany and Switzerland, starting in 1967, peaking in 1968/69, and disappearing after 1972. The mechanism leading to pulmonary hypertension was chronic precapillary vascular obstruction due to plexogenic pulmonary arteriopathy. There was a close geographic as well as temporal relation of the epidemic to the marketing and intake of the appetite depressing drug aminorex fumarate (Menocil). 10 years after the epidemic, half of the patients have died, usually of right heart failure. Of those surviving, half present a definite regression of the pulmonary vascular obstruction. Average survival after the initial diagnosis was 3.5 years in those patients who died. Their PA pressure (+22%) and pulmonary arteriolar resistance (+40%) was higher at the onset of the observation period if compared with the corresponding values of the survivors; also the incidence of right heart failure was significantly higher (84 vs. 58%). Among the surviving patients, the only difference between those with an improved and those with a worsened haemodynamic situation was the age at the beginning of the weight-reducing treatment, those with a progression being 10 years older. The probability of survival after 10 years is considerably higher in chronic pulmonary hypertension of vascular origin (CPHVO) after aminorex than in "classical" primary pulmonary hypertension (CPHVO of unknown cause) and in CPHVO due to recurrent silent pulmonary thromboembolism. This difference in prognosis is an argument in favour of the identity of chronic pulmonary hypertension developing after the intake of the appetite depressing drug aminorex.


A new risk factor for coronary artery disease can be detected by an ApoA1 mAb-based assay.
This study aimed to find coronary artery disease (CAD) related apolipoprotein A1 (ApoA1) monoclonal antibody (mAb) and to evaluate the diagnostic value of the assay based on it. Patients with CAD diagnosed by coronary angiography (disease group, n = 180) and healthy subjects (control group, n = 199) were recruited. The correlation between methods and CAD were evaluated by Spearman's rank correlation coefficients. Receiver operating characteristic (ROC) curve analysis was used to evaluate the auxiliary diagnostic value of methods for CAD. Odds ratios (ORs) of the test results in CAD were estimated using logistic regression analysis. Measurements from an ApoA1 mAb were found significantly positively correlated with CAD (r = 0.243, P < 0.01), unlike the measurements from the ApoA1 pAb were negatively correlated with CAD (r = -0.341, P < 0.001). The areas under the ROC curve of the ApoA1 mAb and pAb measurements were 0.704 and 0.563, respectively, in patients with normal HDL-C levels. ApoA1 values from the mAb assay had a significant positive impact on CAD risk. An ApoA1 mAb-based assay can distinguish a high-density lipoprotein (HDL) subclass positively related to CAD, which can be used to improve and reappraise CAD risk assessment.


The brain metabolic patterns of clozapine- and fluphenazine-treated female patients with schizophrenia: evidence of a sex effect.
The regional cerebral glucose metabolic rates of clozapine-treated and fluphenazine-treated women with schizophrenia and normal controls were obtained by positron emission tomography (PET) using [18F]-2-fluoro-2-deoxy-D-glucose (FDG) as the tracer. The regional metabolic patterns were compared to each other and to the changes previously observed in men. In women, as in men, both clozapine- and fluphenazine-treatment were associated with lower metabolism in the superior prefrontal cortex and higher metabolism in the medial temporal lobe. In both men and women, clozapine treatment led to a greater lowering of inferior prefrontal cortex activity than fluphenazine, which was statistically significant in the larger male cohort. Fluphenazine led to higher metabolic rates in the lateral temporal lobe than clozapine did, but the differences between the two neuroleptics were not statistically significant in either group. The greatest differences in the female as compared to the male responses to fluphenazine and clozapine were in the cingulate and striatum. As compared to controls, the cingulate metabolic rates of women were reduced by 9.1% and 11.4% on clozapine and fluphenazine, respectively; whereas, men have a statistically nonsignificant reduction of 0.1% with clozapine and a 3.2% increase with fluphenazine. In men, fluphenazine was associated with a much greater elevation in basal ganglia metabolic rates than was clozapine, 23.5% as compared to 3.75%; whereas, in women, basal ganglia metabolic rates are nearly equally increased by fluphenazine (21.6%) and clozapine (15.1%).


[Comparative studies of carotid endarterectomy and carotid stenting in patients with coronary artery stenosis (part I)].
Carotid artery stenosis has over the last decade been treated increasingly more often by means of angioplasty and stenting, with the indications for these interventions being gradually broadening. The authors review and analyse the results of multicenter randomized studies and registers regarding carotid artery stent grafting and carotid endarterectomy (CAVATAS, SAPPHIRE, ARCHeR, EVA-3S, CaRESS, SPACE, BEACH, CABERNET, CREATE Pivotal). Presented herein are the outcomes of management of symptomatic and symptom-free patients running both usual and high surgical risks for carotid endarterectomy. The commonly accepted criteria for surgical risks are conventionally subdivided into the absolute and relative ones (i.e., correction-prone). Carotid stenting turned out to be superior to carotid endarterectomy in patients running a high surgical risk.


Extracranial carotid artery in patients with transient global amnesia: evaluation by real-time B-mode ultrasonography with duplex Doppler flow.
Patients with transient global amnesia are often categorized as having cerebrovascular disease. Noninvasive carotid artery testing was performed in 56 patients with transient global amnesia to determine if they had the same incidence of extracranial atherosclerotic vascular disease as patients with focal cerebral transient ischemic attacks. Only 3 of 56 patients had hemodynamic obstruction of flow at the carotid artery bifurcation, and 41 of 56 had no evidence of any atherosclerotic disease. Other risk factors for cerebrovascular disease were present in 24 of 56 patients, but only 1 had a prior cerebrovascular event. The pathophysiology of transient global amnesia appears to differ from the pathophysiology of classical transient ischemic attacks in that there is no clear relation between transient global amnesia and the presence of extracranial atherosclerotic cerebrovascular disease.


Cr(CO)6 photochemistry: semi-classical study of UV absorption spectral intensities and dynamics of photodissociation.
The UV absorption spectrum of Cr(CO)(6) (chromium hexacarbonyl) in gas phase is investigated by theoretical methods with focus on the absorption intensities. It is shown that in spite of good predictions for the excitation energies, the most frequently employed methods for excited-state calculations produce poor predictions for oscillator strengths and absorption cross sections. In particular, time-dependent DFT predicts relative intensities for the two main spectral bands to be up to five times larger than the experimental results depending on the functional. The best results are obtained by a multireference configuration interaction method based on DFT (DFT/MRCI). Spectral shoulders caused by vibronic-coupling absorption are assigned based on symmetry-restricted spectrum simulations. The dynamics of Cr(CO)(6) photodissociation was also considered at TDDFT/B3LYP level. The estimated time constants for the Cr(CO)(6) relaxation and dissociation are in excellent agreement with experimental values. The time constant for internal conversion, however, is longer than the experimentally observed by factor 2, presumably due to an underestimation of the experimental analysis.


Urinary metabolites of organophosphate flame retardants and their variability in pregnant women.
Organophosphate flame retardants (OPFRs) are commonly added to consumer products to reduce their flammability. Based on levels of OPFRs in indoor environments, human exposure is likely chronic and ubiquitous. Animal studies suggest that exposure to some OPFRs may result in adverse health impacts, particularly for Tris (1,3-dichloropropyl) phosphate (TDCPP); however, human data on the impacts of exposure to OPFRs are lacking. To design human studies, more information is needed on the stability of measured OPFRs in human samples over time. In this study, we sought to assess the degree of temporal variability of urinary TDCPP and triphenyl phosphate (TPP) metabolites throughout pregnancy in a cohort of women from central North Carolina. Eight pregnant women provided multiple urine samples: 3 during the 18th week of pregnancy, 1 during the 28th week, and 1 shortly after the child's birth. Bis (1,3-dichloropropyl) phosphate (BDCPP) and diphenyl phosphate (DPP), the respective metabolites of TDCPP and TPP, were measured in urine samples using liquid chromatography-tandem mass spectrometry. BDCPP and DPP were each detected in 38 of 39 urine samples and were not normally distributed. Geometric mean BDCPP and DPP concentrations were 1.3ng/mL (interquartile range (IQR): 0.8, 2.7ng/mL) and 1.9ng/mL (IQR: 0.9, 3.5ng/mL), respectively. BDCPP and DPP were moderately to strongly reliable over one week (intraclass correlation coefficient (ICC)=0.5; 95% confidence interval (CI): 0.4, 0.7 and ICC=0.7; 95% CI: 0.5, 0.8, respectively), and over the entire pregnancy (ICC=0.5 95% CI: 0.3, 0.7 and ICC=0.6; 95% CI: 0.4, 0.7, respectively). These data suggest that exposures to TDCPP and TPP are widespread and variable for pregnant women, and that a single measure of BDCPP or DPP, taken in the second trimester, likely captures information on the rank order of exposure throughout pregnancy.


Oxygen isotope constraints on the sulfur cycle over the past 10 million years.
Oxygen isotopes in marine sulfate (delta18O(SO4)) measured in marine barite show variability over the past 10 million years, including a 5 per mil decrease during the Plio-Pleistocene, with near-constant values during the Miocene that are slightly enriched over the modern ocean. A numerical model suggests that sea level fluctuations during Plio-Pleistocene glacial cycles affected the sulfur cycle by reducing the area of continental shelves and increasing the oxidative weathering of pyrite. The data also require that sulfate concentrations were 10 to 20% lower in the late Miocene than today.


Cardiac Tissue Vascularization: From Angiogenesis to Microfluidic Blood Vessels.
Myocardial infarction results from a blockage of a major coronary artery that shuts the delivery of oxygen and nutrients to a region of the myocardium, leading to massive cardiomyocytes death and regression of microvasculature. Growth factor and cell delivery methods have been attempted to revascularize the ischemic myocardium and prevent further cell death. Implantable cardiac tissue patches were engineered to directly revascularize as well as remuscularize the affected muscle. However, inadequate vascularization in vitro and in vivo limits the efficacy of these new treatment options. Breakthroughs in cardiac tissue vascularization will profoundly impact ischemic heart therapies. In this review, we discuss the full spectrum of vascularization approaches ranging from biological angiogenesis to microfluidic blood vessels as related to cardiac tissue engineering.


Validation of the BATT score for prehospital risk stratification of traumatic haemorrhagic death: usefulness for tranexamic acid treatment criteria.
Tranexamic acid reduces surgical blood loss and reduces deaths from bleeding in trauma patients. Tranexamic acid must be given urgently, preferably by paramedics at the scene of the injury or in the ambulance. We developed a simple score (Bleeding Audit Triage Trauma score) to predict death from bleeding. We conducted an external validation of the BATT score using data from the UK Trauma Audit Research Network (TARN) from 1st January 2017 to 31st December 2018. We evaluated the impact of tranexamic acid treatment thresholds in trauma patients. We included 104,862 trauma patients with an injury severity score of 9 or above. Tranexamic acid was administered to 9915 (9%) patients. Of these 5185 (52%) received prehospital tranexamic acid. The BATT score had good accuracy (Brier score = 6%) and good discrimination (C-statistic 0.90; 95% CI 0.89-0.91). Calibration in the large showed no substantial difference between predicted and observed death due to bleeding (1.15% versus 1.16%, P = 0.81). Pre-hospital tranexamic acid treatment of trauma patients with a BATT score of 2 or more would avoid 210 bleeding deaths by treating 61,598 patients instead of avoiding 55 deaths by treating 9915 as currently. The BATT score identifies trauma patient at risk of significant haemorrhage. A score of 2 or more would be an appropriate threshold for pre-hospital tranexamic acid treatment.


Autophagy Inhibition to Augment mTOR Inhibition: a Phase I/II Trial of Everolimus and Hydroxychloroquine in Patients with Previously Treated Renal Cell Carcinoma.
Everolimus inhibits the mTOR, activating cytoprotective autophagy. Hydroxychloroquine inhibits autophagy. On the basis of preclinical data demonstrating synergistic cytotoxicity when mTOR inhibitors are combined with an autophagy inhibitor, we launched a clinical trial of combined everolimus and hydroxychloroquine, to determine its safety and activity in patients with clear-cell renal cell carcinoma (ccRCC). Three centers conducted a phase I/II trial of everolimus 10 mg daily and hydroxychloroquine in patients with advanced ccRCC. The objectives were to determine the MTD of hydroxychloroquine with daily everolimus, and to estimate the rate of 6-month progression-free survival (PFS) in patients with ccRCC receiving everolimus/hydroxychloroquine after 1-3 prior treatment regimens. Correlative studies to identify patient subpopulations that achieved the most benefit included population pharmacokinetics, measurement of autophagosomes by electron microscopy, and next-generation tumor sequencing. No dose-limiting toxicity was observed in the phase I trial. The recommended phase II dose of hydroxychloroquine 600 mg twice daily with everolimus was identified. Disease control [stable disease + partial response (PR)] occurred in 22 of 33 (67%) evaluable patients. PR was observed in 2 of 33 patients (6%). PFS ≥ 6 months was achieved in 15 of 33 (45%) of patients who achieved disease control. Combined hydroxychloroquine 600 mg twice daily with 10 mg daily everolimus was tolerable. The primary endpoint of >40% 6-month PFS rate was met. Hydroxychloroquine is a tolerable autophagy inhibitor in future RCC or other trials.


Cloning and expression of a new HOXC6 transcript encoding a repressing protein.
Homeodomain-containing proteins are transcription factors that regulate the co-ordinated expression of multiple genes involved in development, differentiation and malignant transformation. In an attempt to characterize expressed homeobox (HOX) genes in breast cancer cells, we cloned two distinct HOXC6 transcripts from an MCF7 cDNA library, Interestingly, one of them represents a new HOXC6 mRNA encoding a homeodomain-containing protein harbouring a unique N-terminal sequence. Moreover we demonstrate that this HOXC6 transcript is less abundant in human breast cancer cells than in non-tumorigenic cell lines, is detected in breast carcinomas and adjacent tissues and is expressed in a variety of human tumours. In addition, transient co-transfection experiments illustrated that both HOXC6 transcripts encode gene products that repress transcription from a HOX binding sequence in MDA-MB231 cells and co-operate with other HOX gene products such as HOXB7 on their target genes. Taken together, our results suggest that HOXC6 proteins might contribute to the breast cell phenotype through co-operative interactions with other HOX-derived proteins and repression of their target genes.


The significance of YKL-40 protein in liver fibrosis.
The aims of this review are to describe the present knowledge about YKL-40 protein, discuss its relation to liver fibrosis, and to look ahead at future perspectives of YKL-40 research. Liver fibrosis is characterized by excess collagen deposition, decreased extracellular matrix degradation and activation of hepatic stellate cells. Therefore, advancement in the identification of liver fibrosis biomarkers with diagnostic and prognostic values becomes an important tool for future molecular therapy. The molecular basis of YKL-40 in liver fibrosis is unknown. A PubMed database search was performed for studies of YKL-40 in liver injury and fibrosis. YKL-40 is an inflammatory glycoprotein involved in endothelial dysfunction by promoting chemotaxis, cell attachment and migration, reorganization, and tissue remodeling as a response to endothelial damage. Several studies demonstrate that elevated serum YKL-levels are independently associated with the presence of endothelial damage and even higher YKL-40 levels are documented in liver fibrosis. YKL-40 may play a key role in liver injury and fibrosis.


Trigger mechanism for the catalytic hydrogen activation by monoiron (iron-sulfur cluster-free) hydrogenase.
A fully optimized model for the resting state of the active site of the third type of phylogenetically unrelated (monoiron) hydrogenase, iron-sulfur cluster-free hydrogenase (Hmd), was constructed based on density functional calculations. This resting state structure shows good agreement with the experimental IR spectra. The calculations predict that the barrier for H2 cleavage in the presence of MPT+ is 18 kcal/mol lower than that in the absence of MPT+, a result that explains why the isotopic H2/D2O exchange catalyzed by Hmd is strictly dependent on the presence of MPT+. This difference is a result of the MPT+ triggering the pyridone to provide electron density to allow the Fe to take a proton while transferring a hydride to the MPT+. These active site models and catalytic mechanism are useful in understanding this hydrogen activation for the design of novel hydrogenation catalysts and for low cost, high efficiency hydrogen generation.


Ultrastructure of the subacromial bursa in painful shoulder syndromes.
In order to determine if inflammation of the subacromial bursa or "bursitis" is as common an occurrence as believed in painful conditions of the shoulder, eight bursae were obtained during surgery from 3 cases of calcifying tendinitis, 2 of tight coracoacromial ligament, 2 of rotator cuff tear and 1 of acromioclavicular osteoarthritis. In the cases of calcifying tendinitis, tight coracoacronial ligament and rotator cuff tear the morphological changes mainly consisted of a numerical increase in cells throughout the bursal wall along with proliferation of endothelial cells in the vascular channels. All of the cells including the endothelial had densely packed, intermediate type filaments in their cytoplasm but no appreciable diminution of metabolic organelles. Lipid droplets were abundant in the extracellular connective tissue of the bursae from the cases with rotator cuff tear. In contrast to the other cases, the bursa from the case of acromioclavicular osteoarthritis showed widespread fibrin deposition in association with cell necrosis. In none of the cases did inflammatory leukocytic cells infiltrate bursal tissue. We conclude, that the subacromial bursa tends to undergo proliferative or degenerative changes in rotator cuff tendinopathies but bursal inflammation with polymorphonuclear cell infiltrate does not occur commonly.


A prospective study of passive smoking and coronary heart disease.
Several epidemiological studies have suggested an association of passive smoking with coronary heart disease (CHD). However, few studies have taken account of exposure to passive smoking in the workplace. Additionally, several studies have been unable to control for the full range of potential confounding factors. We examined prospectively the relationship of passive smoking with risk of CHD in a cohort of women. The study was carried out in an ongoing prospective cohort of US female nurses, in whom we assessed exposure to passive smoking at home and at work as well as duration of years spent living with someone who smoked regularly. We studied 32046 women 36 to 61 years of age in 1982 who had never smoked and were free of diagnosed CHD, stroke, and cancer. During 10 years of follow-up (1982 to 1992), 152 incident cases of CHD (127 nonfatal myocardial infarction and 25 fatal CHD) occurred. Compared with women not exposed to passive smoking, the relative risks of total CHD-adjusted for a broad range of cardiovascular risk factors-were 1.58 (95% CI, 0.93 to 2.68) among those reporting occasional exposure and 1.91 (95% CI, 1.11 to 3.28) among women reporting regular exposure to passive smoking at home or work. There was no relation apparent between duration of living with a smoker and risk of CHD. Despite the fact that exposure to passive smoking was assessed by self-report and only at baseline (as well as other limitations), these data suggest that regular exposure to passive smoking at home or work increases the risk of CHD among nonsmoking women.


Technical note: Evaluation of odor from vaginal discharge of cows in the first 10 days after calving by olfactory cognition and an electronic device.
The objective of this study was to determine test characteristics (i.e., intra- and interobserver variability, intraassay variability, sensitivity, and specificity) of an evaluation of odor from vaginal discharge (VD) of cows in the first 10 d postpartum conducted by olfactory cognition and an electronic device, respectively. In experiment 1, 16 investigators (9 veterinary students and 7 licensed veterinarians) evaluated 5 VD samples each on 10 different days. The kappa test revealed an agreement between investigators (interobserver) of κ=0.43 with a Fleiss adjusted standard error of 0.0061. The overall agreement was the same for students (κ=0.28) and veterinarians (κ=0.28). Mean agreement within observers (intraobserver) was κ=0.52 for all observers, and 0.49 and 0.62 for students and veterinarians, respectively. In experiment 2, the repeatability of an electronic device (DiagNose; C-it, Zutphen, the Netherlands) was tested. Therefore, 5 samples of VD from 5 cows were evaluated 10 times each. The repeatability was 0.97, determined by Cronbach's α. In experiment 3, 20 samples collected from healthy cows and 20 of cows with acute puerperal metritis were evaluated by the 16 investigators and the DiagNose using a dichotomous scale (1=cow with acute puerperal metritis; 0=healthy cow). Sensitivity and specificity of olfactory evaluation was 75.0 and 60.1% compared with 92.0 and 100%, respectively, for the electronic nose device. The study revealed a considerable subjectivity of the human nose concerning the classification into healthy and sick animals based on the assessment of vaginal discharge. The repeatability of the electronic nose was higher. In conclusion, the DiagNose system, although imperfect, is a reasonable tool to improve odor assessment of VD. The current system, however, is not suitable as a screening tool in the field. Further research is warranted to adapt such electronic devices to practical on-farm screening tools.


Genotypes of the UCP1 gene polymorphisms and cardiometabolic diseases: A multifactorial study of association with disease probability.
Cardiometabolic diseases (CMDs) are complex disorders with a heterogenous phenotype, which are caused by multiple factors including genetic factors. Single nucleotide polymorphisms (SNPs) rs45539933 (p.Ala64Thr), rs10011540 (c.-112A>C), rs3811791 (c.-1766A>G), and rs1800592 (c.-3826A>G) in the UCP1 gene have been analyzed for association with CMDs in many studies providing controversial results. However, previous studies only considered individual UCP1 SNPs and did not evaluate them in an integrated manner, which is a more powerful approach to uncover genetic component of complex diseases. This study aimed to investigate associations between UCP1 genotype combinations and CMDs or CMD risk factors in the context of non-genetic factors. We performed multiple logistic regression analysis and proposed new methodology of testing different combinations of SNP genotypes. We found that probability of CMDs increased in presence of the three-SNP combination of genotypes with minor alleles of c.-3826A>G and p.Ala64Thr and wild allele of c.-112A>C, with increasing age, body mass index (BMI), body fat percentage (BF%) and may differ between sexes and between countries. The combination of genotypes with c.-3826A>G minor allele and wild homozygotes of c.-112A>C and p.Ala64Thr was associated with increased probability of diabetes. While combination of genotypes with minor alleles of all three SNPs reduced the CMD probability. The present results suggest that age, BMI, sex, and UCP1 three-SNP combinations of genotypes significantly contribute to CMD probability. Varying of c.-112A>C alleles in the genotype combination with minor alleles of c.-3826A>G and p.Ala64Thr markedly changes CMD probability.


Unnexins: Homologs of innexin proteins in Trypanosomatidae parasites.
Large-pore channels, including those formed by connexin, pannexin, innexin proteins, are part of a broad family of plasma membrane channels found in vertebrates and invertebrates, which share topology features. Despite their relevance in parasitic diseases such as Chagas and malaria, it was unknown whether these large-pore channels are present in unicellular organisms. We identified 14 putative proteins in Trypanosomatidae parasites as presumptive homologs of innexin proteins. All proteins possess the canonical motif of the innexin family, a pentapeptide YYQWV, and 10 of them share a classical membrane topology of large-pore channels. A sequence similarity network analysis confirmed their closeness to innexin proteins. A bioinformatic model showed that a homolog of Trypanosoma cruzi (T. cruzi) could presumptively form a stable octamer channel with a highly positive electrostatic potential in the internal cavities and extracellular entrance due to the notable predominance of residues such as Arg or Lys. In vitro dye uptake assays showed that divalent cations-free solution increases YO-PRO-1 uptake and hyperosmotic stress increases DAPI uptake in epimastigotes of T. cruzi. Those effects were sensitive to probenecid. Furthermore, probenecid reduced the proliferation and transformation of T. cruzi. Moreover, probenecid or carbenoxolone increased the parasite sensitivity to antiparasitic drugs commonly used in therapy against Chagas. Our study suggests the existence of innexin homologs in unicellular organisms, which could be protein subunits of new large-pore channels in unicellular organisms.


Involvement of pituitary gonadotropins, gonadal steroids and breeding season in sex change of protogynous dusky grouper, Epinephelus marginatus (Teleostei: Serranidae), induced by a non-steroidal aromatase inhibitor.
Two experiments were performed using the aromatase inhibitor (AI) letrozole (100mg/kg) to promote sex change, from female-to-male, in protogynous dusky grouper. One experiment was performed during the breeding season (spring) and the other at the end of the breeding season (summer). During the spring, AI promoted sex change after 9 weeks and the sperm produced was able to fertilize grouper oocytes. During the summer, the sex change was incomplete; intersex individuals were present and sperm was not released by any of the animals. Sex changed gonads had a lamellar architecture; cysts of spermatocytes and spermatozoa in the lumen of the germinal compartment. In the spring, after 4 weeks, 11ketotestosterone (11KT) levels were higher in the AI than in control fish, and after 9 weeks, coincident with semen release, testosterone levels increased in the AI group, while 11KT returned to the initial levels. Estradiol (E2) levels remained unchanged during the experimental period. Instead of decreasing throughout the period, as in control group, 17 α-OH progesterone levels did not change in the AI-treated fish, resulting in higher values after 9 weeks when compared with control fish. fshβ and lhβ gene expression in the AI animals were lower compared with control fish after 9 weeks. The use of AI was effective to obtain functional males during the breeding season. The increase in androgens, modulated by gonadotropins, triggered the sex change, enabling the development of male germ cells, whereas a decrease in E2 levels was not required to change sex in dusky grouper.


18F-FDG PET image biomarkers improve prediction of late radiation-induced xerostomia.
Current prediction of radiation-induced xerostomia 12months after radiotherapy (Xer12m) is based on mean parotid gland dose and baseline xerostomia (Xerbaseline) scores. The hypothesis of this study was that prediction of Xer12m is improved with patient-specific characteristics extracted from 18F-FDG PET images, quantified in PET image biomarkers (PET-IBMs). Intensity and textural PET-IBMs of the parotid gland were collected from pre-treatment 18F-FDG PET images of 161 head and neck cancer patients. Patient-rated toxicity was prospectively collected. Multivariable logistic regression models resulting from step-wise forward selection and Lasso regularisation were internally validated by bootstrapping. The reference model with parotid gland dose and Xerbaseline was compared with the resulting PET-IBM models. High values of the intensity PET-IBM (90th percentile (P90)) and textural PET-IBM (Long Run High Grey-level Emphasis 3 (LRHG3E)) were significantly associated with lower risk of Xer12m. Both PET-IBMs significantly added in the prediction of Xer12m to the reference model. The AUC increased from 0.73 (0.65-0.81) (reference model) to 0.77 (0.70-0.84) (P90) and 0.77 (0.69-0.84) (LRHG3E). Prediction of Xer12m was significantly improved with pre-treatment PET-IBMs, indicating that high metabolic parotid gland activity is associated with lower risk of developing late xerostomia. This study highlights the potential of incorporating patient-specific PET-derived functional characteristics into NTCP model development.


Is there any gender or age-related discrepancy in the waiting time for each step in the surgical management of acute traumatic cervical spinal cord injury?
Context/Objective: Prior studies indicate that patient's gender and age can influence treatment choices during spine disease management. This study examines whether individual's gender and age at injury onset influence the waiting time for each step in the surgical management of patients with acute traumatic cervical spinal cord injury (atcSCI). Design: Retrospective cohort study. Setting: Quaternary spine trauma center. Participants: This study included consecutive individuals with atcSCI admitted from August/2002 to October/2008 who were enrolled in the Surgical Trial in Acute Spinal Cord Injury Study (STASCIS). Interventions: Spinal cord decompression. Outcome Measures: Data on the periods of time for each step in the surgical management were analyzed to explore the potential effects of gender and age at injury onset. Results: There were 64 individuals with atcSCI (17 women, 47 men; age range: 18-78 years; mean age: 50.5 ± 2.1 years). Older age was associated with longer stay in the acute spine center, but this association was cofounded by major pre-existing medical co-morbidities. Age did not significantly affect the waiting time for each step in the surgical management of these individuals with atcSCI. Women underwent surgical assessment earlier than men. Gender did not influence other key steps in the surgical management. Conclusion: The study results suggest that older age at injury onset was associated with longer stay in the acute spine care center, and women had a shorter waiting time for surgical assessment than men. Nevertheless, no other age or gender bias was identified in the waiting times for the steps in the management of atcSCI.


Efficacy of personalized cognitive counseling in men of color who have sex with men: secondary data analysis from a controlled intervention trial.
In a previous report, we demonstrated the efficacy of a cognitively based counseling intervention compared to standard counseling at reducing episodes of unprotected anal intercourse (UAI) among men who have sex with men (MSM) seeking HIV testing. Given the limited number of efficacious prevention interventions for MSM of color (MOC) available, we analyzed the data stratified into MOC and whites. The sample included 196 white MSM and 109 MOC (23 African Americans, 36 Latinos, 22 Asians, eight Alaskan Natives/Native Americans/Hawaiian/Pacific Islander, and 20 of mixed or other unspecified race). Among MOC in the intervention group, the mean number of episodes of UAI declined from 5.1 to 1.6 at six months and was stable at 12 months (1.8). Among the MOC receiving standard counseling, the mean number of UAI episodes was 4.2 at baseline, 3.9 at six months and 2.1 at 12 months. There was a significant treatment effect overall (relative risk 0.59, 95% confidence interval 0.35-0.998). These results suggest that the intervention is effective in MOC.


Heightened self-reported punishment sensitivity, but no differential attention to cues signaling punishment or reward in anorexia nervosa.
This study examined whether adolescents with anorexia nervosa (AN) are more sensitive to punishment and less sensitive to reward than a non-eating disorder comparison group. Both self-report and performance measures were used to index reward and punishment sensitivity. Participants were adolescents with AN (n = 69) and an individually matched comparison group with healthy weight (n = 69). They completed the Behavioral Inhibition Scale/Behavioral Activation Scale and the Sensitivity to Punishment and Sensitivity to Reward Questionnaire to index self-reported reward and punishment sensitivity, and performed the Spatial Orientation Task to index attention to cues signaling reward and punishment. There was extremely strong evidence (BF10 > 100), that adolescents with AN reported higher sensitivity to punishment than adolescents without an eating disorder. However, adolescents with AN did not differ from the comparison group on self-reported reward sensitivity, and attention to cues signaling reward or punishment. Adolescents with AN clearly show heightened punishment sensitivity, yet this was not paralleled by a heightened proneness to detect signals of punishment. An important next step would be to examine whether punishment sensitivity is a reliable risk factor for the development or maintenance of AN.


Ferruginol suppresses survival signaling pathways in androgen-independent human prostate cancer cells.
Ferruginol, a bioactive compound isolated from a Chilean tree (Podocarpaceae), attracts attention as a consequence of its pharmacological properties, which include anti-fungal, anti-bacterial, cardioprotective, anti-oxidative, anti-plasmodial and anti-ulcerogenic actions. Nevertheless, the molecular basis for these actions remains only partly understood and hence we investigated the effects of ferruginol on androgen-independent human prostate cancer cells (PC3), a known model for solid tumor cells with an exceptional resistance to therapy. The results show that ferruginol induces PC3 cell death via activation of caspases as well as apoptosis-inducing factor (AIF) as confirmed by its translocation into the nucleus. In order to clarify the biochemical mechanism responsible for the anti-tumor activity of ferruginol, we analyzed a set of molecular mediators involved in tumor cell survival, progression and aggressiveness. Ferruginol was able to trigger inhibition/downregulation of Ras/PI3K, STAT 3/5, protein tyrosine phosphatase and protein kinases related to cell cycle regulation. Importantly, the toxic effect of ferruginol was dramatically impeded in a more reducing environment, which indicates that at least in part, the anti-tumoral activity of ferruginol might be related to redox status modulation. This study supports further examination of ferruginol as a potential agent for both the prevention and treatment of prostate cancer.


Acute abdominal pain due to biliary cystadenoma.
We present a case of a biliary cystadenoma, a rare benign cystic tumor arising in most cases of the intrahepatic bile ducts. A 30-year old woman presented with abdominal pain in the right upper quadrant for about 10 days with increasing severity. Radiological evaluation by means of abdominal ultrasound and CT-scan revealed a multilocular cystic lesion in the right liver lobe, not present on abdominal ultrasound performed 10 years before. The diagnosis of a biliary cystadenoma was proposed and complete surgical resection of the mass was performed.


Expression profiling of lymphocyte plasma membrane proteins.
The physicochemical properties of plasma membrane proteins of mammalian cells render them refractory to systematic analysis by two-dimensional electrophoresis. We have therefore used in vivo cell surface labeling with a water-soluble biotinylation reagent, followed by cell lysis and membrane purification, prior to affinity capture of biotinylated proteins. Purified membrane proteins were then separated by solution-phase isoelectric focusing and SDS-PAGE and identified by high-pressure liquid chromatography electrospray/tandem mass spectrometry. Using this approach, we identified 42 plasma membrane proteins from a murine T cell hybridoma and 46 from unfractionated primary murine splenocytes. These included three unexpected proteins; nicastrin, osteoclast inhibitory lectin, and a transmembrane domain-containing hypothetical protein of 11.4 kDa. Following stimulation of murine splenocytes with phorbol ester and calcium ionophore, we observed differences in expression of CD69, major histocompatibility complex class II molecules, the glucocorticoid-induced TNF receptor family-related gene product, and surface immunoglobulin M and D that were subsequently confirmed by Western blot or flow cytometric analysis. This approach offers a generic and powerful strategy for investigating differential expression of surface proteins in many cell types under varying environmental and pathophysiological conditions.


The SR-mitochondria interaction: a new player in cardiac pathophysiology.
Mitochondria are essential for energy supply and cell signalling and may be triggers and effectors of cell death. Mitochondrial respiration is tightly controlled by the matrix Ca(2+) concentration, which is beat-to-beat regulated by uptake and release mainly through the mitochondrial Ca(2+) uniporter and Na(+)/Ca(2+) exchanger, respectively. Recent studies demonstrate that mitochondrial Ca(2+) uptake is more dependent on anatomo-functional microdomains established with the sarcoplasmic reticulum (SR) than on cytosolic Ca(2+). This privileged communication between SR and mitochondria is not restricted to Ca(2+) but may involve ATP and reactive oxygen species, which has important implications in cardiac pathophysiology. The disruption of the SR-mitochondria interaction caused by cell remodelling has been implicated in the deterioration of excitation-contraction coupling of the failing heart. The SR-mitochondria interplay has been suggested to be involved in the depressed Ca(2+) transients and mitochondrial dysfunction observed in diabetic hearts as well as in the genesis of certain arrhythmias, and it may play an important role in myocardial reperfusion injury. During reperfusion, re-energization in the presence of cytosolic Ca(2+) overload results in SR-driven Ca(2+) oscillations that may promote mitochondrial permeability transition (MPT). The relationship between MPT and Ca(2+) oscillations is bidirectional, as recent data show that the induction of MPT in Ca(2+)-overloaded cardiomyocytes may result in mitochondrial Ca(2+) release that aggravates Ca(2+) handling and favours hypercontracture. A more complete characterization of the structural arrangements responsible for SR-mitochondria interplay will allow better understanding of cardiac (patho)physiology but also, and no less important, should serve as a basis for the development of new treatments for cardiac diseases.


Synthesis of catalytic materials in flames: opportunities and challenges.
The proven capacity of flame aerosol technology for rapid and scalable synthesis of functional nanoparticles makes it ideal for the manufacture of an array of heterogeneous catalysts. Capitalizing on the high temperature environment, rapid cooling and intimate component mixing at either atomic or nano scale, novel catalysts with unique physicochemical properties have been made using flame processes. This tutorial review covers the main features of flame synthesis and illustrates how the physical and chemical properties of as-synthesized solid catalytic materials can be controlled by proper choice of the process parameters. Gas phase particle formation mechanisms and the effect of synthesis conditions (reactor configuration, precursor and dispersion gas flow rates, temperature and concentration fields) on the structural, chemical and catalytic properties of as-prepared materials are discussed. Finally, opportunities and challenges offered by flame synthesis of catalytic materials are addressed.


Probability learning and feedback processing in dyslexia: A performance and heart rate analysis.
Recent studies suggest that individuals with dyslexia may be impaired in probability learning and performance monitoring. These observations are consistent with findings indicating atypical neural activations in frontostriatal circuits in the brain, which are important for associative learning. The current study further examined probability learning and performance monitoring in adult individuals with dyslexia (n = 23) and typical readers (n = 31) using two varieties of a typical probabilistic learning task. In addition to performance measures, we measured heart rate, focusing on cardiac slowing with negative feedback as a manifestation of the automatic performance monitoring system. One task required participants to learn associations between artificial script and speech sounds and the other task required them to learn associations between geometric forms and bird sounds. Corrective feedback (informative or random) was provided in both tasks. Performance results indicated that individuals with dyslexia and typical readers learned the associations equally well in contrast to expectations. We found the typical cardiac response associated with feedback processing consisting of a heart rate slowing with the presentation of the feedback and a return to baseline thereafter. Interestingly, the heart rate slowing associated with feedback was less pronounced and the return to baseline was delayed in individuals with dyslexia relative to typical readers. These findings were interpreted in relation to current theorizing of performance monitoring linking the salience network in the brain to autonomic functioning.


Thermal Injury Deaths of Community-dwelling Older People With Dementia.
Although people with dementia are prone to the risk of fire and burn injuries, the relationship between fatal thermal injuries and dementia has not been investigated. We examined coronial files in the state of Victoria, Australia between July 1, 2000 and December 31, 2014, for unintentional thermal injury causing deaths of community dwelling people, aged 65 years and older to identify those with dementia. Mortality for Victorian populations with and without dementia was calculated using direct age standardization. In total, 105 deaths were identified of which 18 (17%) had dementia. Mortality was more than 3-fold greater for people with dementia (3.0 vs. 0.8 per 100,000 person-years) and they more often lived with others when the injury occurred (56%, n=10 vs. 29% n=25, P=0.028). Our results suggest that older people with dementia are at greater risk of death through thermal injury. Further research should consider investigating a larger population through combining databases across multiple jurisdictions.


Pediatric Guillain-Barré syndrome: Indicators for a severe course.
This study aims to retrospectively evaluate pediatric Guillain-Barré syndrome cases in a tertiary center in Istanbul, Turkey. The data of 40 patients with Guillain-Barré syndrome who had been admitted to the Department of Pediatrics at the Istanbul University Medical Faculty between 2005 and 2011 were collected. Mann-Whitney U, Kruskal-Wallis, chi-square, and Fisher's exact tests were used for statistical analysis. Mean patient age was 5.4 ± 3.0 years; 20 out of 40 patients (50%) were female and 20 (50%) were male. Preceding infection was detected in 32 cases (80%). Six patients had speech impairment. Out of eight patients with respiratory distress (20%), five required respiratory support (12.5%) of which three of them had speech impairment as well. According to nerve conduction studies, 21 patients (52.5%) had acute inflammatory demyelinating polyradiculoneuropathy, 14 (35%) had acute motor axonal neuropathy, and five (12.5%) had acute motor-sensory axonal neuropathy. Thirty-three patients (82.5%) received intravenous immunglobulin, 3 (7.5%) underwent plasmapheresis and 4 (10%) received both. Time until recovery (P = 0.022) and time until aided (P = 0.036) and unaided (P = 0.027) walking were longer in patients with acute gastrointestinal infection than in those with upper respiratory tract infection (P < 0.05). Time until response to treatment (P = 0.001), time until aided (P = 0.001) and unaided (P = 0.002) walking, and time until complete recovery (P = 0.002) were longer in acute motor axonal neuropathy cases as compared to acute inflammatory demyelinating polyradiculoneuropathy cases. Recovery was longer with acute gastrointestinal infection and acute motor axonal neuropathy. Speech impairment could be a clinical clue for the need of mechanical ventilation.


Educational History, Employment Characteristics, and Desired Competencies of Doctoral-Educated Athletic Trainers.
OBJECTIVE: The study had 3 objectives: (1) to assess the educational history of doctoral-educated certified athletic trainers (ATCs) who work at academic institutions, (2) to determine the current employment characteristics of doctoral-educated ATCs who work at academic institutions, and (3) to identify which competencies doctoral-educated ATCs feel are important for new doctoral graduates to possess upon graduation. DESIGN AND SETTING: Multiple sources were used to identify doctoral-educated ATCs who work at academic institutions. These individuals were surveyed to assess their educational histories, current employment characteristics, and opinions on desired competencies for new doctoral graduates. Data were analyzed using descriptive and inferential statistics. SUBJECTS: Surveys were sent to 130 individuals, and the response rate was 89.2% (n = 116). MEASUREMENTS: Subjects answered questions regarding their educational history and employment characteristics. A 5-point Likert scale was used to assess the importance of 22 competencies for new doctoral graduates to possess upon graduation. Comparisons were made between program directors and non-program directors, respondents employed at doctoral-granting institutions and non-doctoral-granting institutions, and doctoral student advisors and non-advisors. RESULTS: Subjects reported several different educational backgrounds, job titles, and job responsibilities. Significant differences in job responsibilities and assessment of desired competencies were found between program directors and non-program directors, employees of doctoral-granting institutions and non-doctoral-granting institutions, and doctoral student advisors and non-advisors. CONCLUSIONS: As new doctoral programs are established in athletic training, students should receive training as classroom instructors and program administrators, in addition to learning the skills necessary to perform independent research in athletic training.


A systematic review of models to predict recruitment to multicentre clinical trials.
Less than one third of publicly funded trials managed to recruit according to their original plan often resulting in request for additional funding and/or time extensions. The aim was to identify models which might be useful to a major public funder of randomised controlled trials when estimating likely time requirements for recruiting trial participants. The requirements of a useful model were identified as usability, based on experience, able to reflect time trends, accounting for centre recruitment and contribution to a commissioning decision. A systematic review of English language articles using MEDLINE and EMBASE. Search terms included: randomised controlled trial, patient, accrual, predict, enroll, models, statistical; Bayes Theorem; Decision Theory; Monte Carlo Method and Poisson. Only studies discussing prediction of recruitment to trials using a modelling approach were included. Information was extracted from articles by one author, and checked by a second, using a pre-defined form. Out of 326 identified abstracts, only 8 met all the inclusion criteria. Of these 8 studies examined, there are five major classes of model discussed: the unconditional model, the conditional model, the Poisson model, Bayesian models and Monte Carlo simulation of Markov models. None of these meet all the pre-identified needs of the funder. To meet the needs of a number of research programmes, a new model is required as a matter of importance. Any model chosen should be validated against both retrospective and prospective data, to ensure the predictions it gives are superior to those currently used.


Two-part random effects growth modeling to identify risks associated with alcohol and cannabis initiation, initial average use and changes in drug consumption in a sample of adult, male twins.
Our aim was to profile alcohol and cannabis initiation and to characterize the effects of developmental and environmental risk factors on changes in average drug use over time. We fitted a two-part random effects growth model to identify developmental and environmental risks associated with alcohol and cannabis initiation, initial average use and changes in average use. 1796 males aged 24-63 from the Virginia Adult Twin Study of Psychiatric and Substance Use Disorders. Data from three interview waves included self-report measures of average alcohol and cannabis use between ages 15 and 24, genetic risk of problem drug use, childhood environmental risks, personality, psychiatric symptoms, as well as personal, family and social risk factors. Average alcohol and cannabis use were correlated at all ages. Genetic risk of drug use based on family history, higher sensation seeking, and peer group deviance predicted both alcohol and cannabis initiation. Higher drug availability predicted cannabis initiation while less parental monitoring and drug availability were the best predictors of how much cannabis individuals consumed over time. The liability to initiate alcohol and cannabis, average drug use as well as changes in drug use during teenage years and young adulthood is associated with known risk factors.


Transient c-Src Suppression During Endodermal Commitment of Human Induced Pluripotent Stem Cells Results in Abnormal Profibrotic Cholangiocyte-Like Cells.
Directed differentiation of human induced pluripotent stem cells (iPSCs) toward hepatobiliary lineages has been increasingly used as models of human liver development/diseases. As protein kinases are important components of signaling pathways regulating cell fate changes, we sought to define the key molecular mediators regulating human liver development using inhibitors targeting tyrosine kinases during hepatic differentiation of human iPSCs. A library of tyrosine kinase inhibitors was used for initial screening during the multistage differentiation of human iPSCs to hepatic lineage. Among the 80 kinase inhibitors tested, only Src inhibitors suppressed endoderm formation while none had significant effect on later stages of hepatic differentiation. Transient inhibition of c-Src during endodermal induction of human iPSCs reduced endodermal commitment and expression of endodermal markers, including SOX17 and FOXA2, in a dose-dependent manner. Interestingly, the transiently treated cells later developed into profibrogenic cholangiocyte-like cells expressing both cholangiocyte markers, such as CK7 and CK19, and fibrosis markers, including Collagen1 and smooth muscle actin. Further analysis of these cells revealed colocalized expression of collagen and yes-associated protein (YAP; a marker associated with bile duct proliferation/fibrosis) and an increased production of interleukin-6 and tumor necrosis factor-α. Moreover, treatment with verteporfin, a YAP inhibitor, significantly reduced expression of fibrosis markers. In summary, these results suggest that c-Src has a critical role in cell fate determination during endodermal commitment of human iPSCs, and its alteration in early liver development in human may lead to increased production of abnormal YAP expressing profibrogenic proinflammatory cholangiocytes, similar to those seen in livers of patients with biliary fibrosis. Stem Cells 2019;37:306-317.


Xenopus Cdc7 executes its essential function early in S phase and is counteracted by checkpoint-regulated protein phosphatase 1.
The initiation of DNA replication requires two protein kinases: cyclin-dependent kinase (Cdk) and Cdc7. Although S phase Cdk activity has been intensively studied, relatively little is known about how Cdc7 regulates progression through S phase. We have used a Cdc7 inhibitor, PHA-767491, to dissect the role of Cdc7 in Xenopus egg extracts. We show that hyperphosphorylation of mini-chromosome maintenance (MCM) proteins by Cdc7 is required for the initiation, but not for the elongation, of replication forks. Unlike Cdks, we demonstrate that Cdc7 executes its essential functions by phosphorylating MCM proteins at virtually all replication origins early in S phase and is not limiting for progression through the Xenopus replication timing programme. We demonstrate that protein phosphatase 1 (PP1) is recruited to chromatin and rapidly reverses Cdc7-mediated MCM hyperphosphorylation. Checkpoint kinases induced by DNA damage or replication inhibition promote the association of PP1 with chromatin and increase the rate of MCM dephosphorylation, thereby counteracting the previously completed Cdc7 functions and inhibiting replication initiation. This novel mechanism for regulating Cdc7 function provides an explanation for previous contradictory results concerning the control of Cdc7 by checkpoint kinases and has implications for the use of Cdc7 inhibitors as anti-cancer agents.


Postmeningitic pediatric hearing loss from non-type b Haemophilus influenzae.
Postmeningitic hearing loss from Haemophilus influenzae (H. influenzae) is increasingly due to encapsulated serotypes other than type b (Hib) and nontypeable strains (collectively, nHiB H. influenzae). Pediatric hearing loss after nHib H. influenzae meningitis remains poorly described. Retrospecive case series of nHiB H. influenzae meningitis cases identified from a microbiologic database at Children's Hospital Colorado from 2000 to 2020. Literature regarding nHiB H. influenzae and H. influenzae postmeningitic hearing loss was also reviewed. Eleven cases of nHib H. influenzae meningitis (median age 15.9 months) were identified due to serotype f (36 %), serotype a (27 %), and nontypable strains (36 %). Seven (64 %) patients were male, 55 % were white and 18 % were Hispanic or Latino. Hearing loss was initially identified in 4 children (40 %), with two patients with moderate conductive hearing loss (CHL) and one child with unilateral moderate sensorineural (SNHL) hearing loss patients recovering normal hearing. One patient with bilateral profound sensorineural hearing loss and associated labyrinthitis ossificans required cochlear implantation. All children (4) with identified hearing loss were noted to have additional intracranial sequelae, which included empyema (2), sinus thrombosis (2), and seizures (2). Of patients receiving steroids, 25 % had hearing loss on initial testing, compared to 66 % of those who did not receive steroids. nHib H. influenzae can cause both transient and permanent postmeningitic hearing loss. Steroids may offer otoprotection in nHib H. influenzae meningitis similar to Hib meningitis. Given the limited literature, further study is needed to better characterize hearing outcomes after nHib H. influenzae meningitis.


Enhancing patient safety and quality of care by improving the usability of electronic health record systems: recommendations from AMIA.
In response to mounting evidence that use of electronic medical record systems may cause unintended consequences, and even patient harm, the AMIA Board of Directors convened a Task Force on Usability to examine evidence from the literature and make recommendations. This task force was composed of representatives from both academic settings and vendors of electronic health record (EHR) systems. After a careful review of the literature and of vendor experiences with EHR design and implementation, the task force developed 10 recommendations in four areas: (1) human factors health information technology (IT) research, (2) health IT policy, (3) industry recommendations, and (4) recommendations for the clinician end-user of EHR software. These AMIA recommendations are intended to stimulate informed debate, provide a plan to increase understanding of the impact of usability on the effective use of health IT, and lead to safer and higher quality care with the adoption of useful and usable EHR systems.


[Is shortage of medical practitioners only a question of numbers? A phenomenologic approach to the scarcity of physicians in a county].
This paper presents a qualitative study of the employment of medical doctors in Møre and Romsdal County. On the basis of the fact that Norway has more physicians in relation to the population than other Northern European countries, we try to explore the reality behind the excessive demand for physicians? On the basis of interviews with physicians in hospitals and primary health care, we suggest four categories of explanations. First, areas with low population density experience a traditional scarcity of doctors. Second, a scarcity of doctors is locally constructed in municipalities with no excessive demand from patients for medical aid, but with vacant positions. Third, medical science generates a scarcity of doctors as medical specialisation has gone beyond the level needed for work in small hospitals. Fourth, there is a demand-driven scarcity of doctors caused by new treatments generating new patients. The four categories may differentiate the debate on how to solve the Norwegian problem of scarcity of medical doctors.