Trends in the vertical distribution of ozone
Analyses of satellite, ground-based, and balloon measurements allow updated estimates of trends in the vertical profile of ozone since 1979. The results show overall consistency among several independent measurement systems, particularly for northern hemisphere midlatitudes where most balloon and ground-based measurements are made. Combined trend estimates over these latitudes for the period 1979-96 show statistically significant negative trends at all altitudes between 10 and 45 km, with two local extremes: -7.4 +/- 2. 0% per decade at 40 km and -7.3 +/- 4.6% per decade at 15 km altitude. There is a strong seasonal variation in trends over northern midlatitudes in the altitude range of 10 to 18 km, with the largest ozone loss during winter and spring. The profile trends are in quantitative agreement with independently measured trends in column ozone, the amount of ozone in a column above the surface. The vertical profiles of ozone trends provide a fingerprint for the mechanisms of ozone depletion over the last two decades.


Cholinergic properties of soy.
Soybeans have been shown to have numerous health benefits, but the underlying mechanisms are poorly understood. The aim of this study was to characterize some pharmacologic properties of active substances in aqueous soy extract. The pharmacologic actions of the extract were tested by measuring mechanical activity of isolated guinea-pig ileum in an organ bath. The ileum contracted in response to soy extract in a concentration-dependent manner. This response was unaffected by the nerve blocker tetrodotoxin (10(-6) M) but was completely inhibited by atropine (10(-9) M), indicating an action via muscarinic receptors on the muscle. In the presence of the M(3) muscarinic antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide and to a lesser extent in the presence of the M(2) muscarinic antagonist 11-([2-[(diethylamino)methyl]-1-piperidinyl]acetyl)-5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepine-6-one, the response was decreased. When acetylcholine (ACh) esterase inhibitors were added to the medium before the addition of soy extract, the response to the extract was potentiated. Preincubation of the extract with exogenous ACh esterase reduced its activity. The response to choline, ACh, and phosphorylcholine was also tested, and none of these substances accurately replicated the response to soy extract. However, some qualitative similarities were observed between the effect of choline and ACh to that of the extract. These results indicate the presence of an ACh-like substance in soy. Due to the abundance and importance of muscarinic receptors, the presence of a cholinergic substance in soy could have numerous implications. The role of this substance in the beneficial effect of soy on various body systems merits further investigation.


Mechanism of traumatic retinal detachment in blunt impact: a finite element study.
Retinal detachment typically occurs when the retina is pulled away from its normal position by blunt trauma. It has been estimated that traumatic retinal detachments account for 10-20% of all detachments. Understanding the mechanism of traumatic retinal detachment is helpful for ophthalmologists to make a more accurate diagnosis before the symptoms develop. A finite element eye model, validated through published data, was used to simulate traumatic retinal detachment. Retinal adhesive force was incorporated into the model using breakable bonded contact. Under BB impact, global deformation was divided into four stages: compression, decompression, overshooting and oscillation. Shockwave propagation in the retina produced high strain in the retina. For an impact speed of 50 m/s, the peak strain of 0.138 in ora serrata exceeded the specified threshold for retinal break. When the eye was decompressed, negative pressure occurred around and anterior to the equator, with a minimum of -663 kPa, leading to retinal detachment. The following relative inertia motions between the retina and its supporting tissue extended the detachment. In addition, the simulations of lower shear modulus of vitreous and increased retinal adhesive force also confirm that the extent of retinal detachment is determined by negative pressure and inertial motion. In conclusion, shockwave and negative pressure contribute to retinal detachment. Shockwave propagation in the retina leads to retinal break, while negative pressure and relative inertial motion could pull the retina away from the supporting tissue. The current work would help understand the basic mechanisms underlying blunt trauma.


Dynamics of Docosahexaenoic Acid Utilization by Mouse Peritoneal Macrophages.
In this work, the incorporation of docosahexaenoic acid (DHA) in mouse resident peritoneal macrophages and its redistribution within the various phospholipid classes were investigated. Choline glycerophospholipids (PC) behaved as the major initial acceptors of DHA. Prolonged incubation with the fatty acid resulted in the transfer of DHA from PC to ethanolamine glycerophospholipids (PE), reflecting phospholipid remodeling. This process resulted in the cells containing similar amounts of DHA in PC and PE in the resting state. Mass spectrometry-based lipidomic analyses of phospholipid molecular species indicated a marked abundance of DHA in ether phospholipids. Stimulation of the macrophages with yeast-derived zymosan resulted in significant decreases in the levels of all DHA-containing PC and PI species; however, no PE or PS molecular species were found to decrease. In contrast, the levels of an unusual DHA-containing species, namely PI(20:4/22:6), which was barely present in resting cells, were found to markedly increase under zymosan stimulation. The levels of this phospholipid also significantly increased when the calcium-ionophore A23187 or platelet-activating factor were used instead of zymosan to stimulate the macrophages. The study of the route involved in the synthesis of PI(20:4/22:6) suggested that this species is produced through deacylation/reacylation reactions. These results define the increases in PI(20:4/22:6) as a novel lipid metabolic marker of mouse macrophage activation, and provide novel information to understand the regulation of phospholipid fatty acid turnover in activated macrophages.


Glutathione metabolism of the erythrocyte. The enzymic cleavage of glutathione-haemoglobin preparations by glutathione reductase.
A complex of haemoglobin and GSH was prepared by incubating haemoglobin with GSH and acetylphenylhydrazine. GSH could be released from the crude preparation by incubation with NADPH. However, when the haemoglobin preparation was separated from glutathione reductase by DEAE-Sephadex chromatography, NADPH no longer released GSH. Rather, the addition of a combination of either partially purified human erythrocyte or crystalline glutathione reductase and NADPH was required to release GSH from the haemoglobin-GSH complex. This complex is commonly believed to represent a mixed disulphide of GSH and the cysteine-beta-93 thiol group. This interpretation was supported by the finding that prior alkylation of available haemoglobin thiol groups prevented the formation of the complex. By using haemoglobin-[(35)S]GSH complex as a substrate, it was shown that GSH itself released the radioactivity from the complex only very slowly. In contrast, the release of [(35)S]GSH was very rapid in the presence of NADPH and glutathione reductase. This suggests that the cleavage of the haemoglobin-GSH complex is not mediated by GSH with cyclic reduction of GSSG formed, but rather proceeds enzymically through glutathione reductase.


CA125 expression in epithelioid sarcoma.
There has been no report on useful immunohistological markers for epithelioid sarcoma (ES) so far. The purpose of this study is to evaluate the positivity and specificity of CA125 as a marker for the correct diagnosis of ES. This study was performed in 11 patients with ES (nine men and two women; distal type: 10 cases; proximal type: one case), 78 patients with other soft tissue tumors and nine with benign granulomas. The other soft tissue tumors consisted of six synovial sarcomas, six clear cell sarcomas, eight leiomyosarcomas, six rhabdomyosarcomas, five malignant peripheral nerve sheath tumors, ten malignant fibrous histiocytomas, 17 desmoid tumors, 14 liposarcomas, six squamous cell carcinomas (cutaneous SCC of the distal extremities), two rheumatoid nodules and seven foreign body granulomas. Immunohistochemical analysis for CA125 was performed for these 89 soft tissue tumors and nine granulomas using a labeled streptavidin biotin method. Immunohistochemical analysis of epithelial membrane antigen, cytokeratin, carcinoembrionic antigen, vimentin and CD34 was performed only for the 11 ES patients. CA125 was strongly expressed in 10 out of the 11 ES patients. EMA, cytokeratin and vimentin were also positive in all the cases. CEA was positive in two of the 11 patients. Immunohistochemical study in six ES patients showed expression of CD 34. The other 78 soft tissue tumors and nine granulomas did not express CA125. This study clearly revealed the specificity and positivity of CA125 in ES. These data indicate that CA125 may be a useful tumor marker for diagnosing ES.


The effect of the coupled oxidation of substrate on the permeability of blowfly flight-muscle mitochondria to potassium and other cations.
1. Blowfly flight-muscle mitochondria respiring in the absence of phosphate acceptor (i.e. in state 4) take up greater amounts of K(+), Na(+), choline, phosphate and Cl(-) (but less NH(4) (+)) than non-respiring control mitochondria. 2. Uptake of cations is accompanied by an increase in the volume of the mitochondrial matrix, determined with the use of [(14)C]-sucrose and (3)H(2)O. The osmolarity of the salt solution taken up was approximately that of the suspending medium. 3. The [(14)C]sucrose-inaccessible space decreased with increasing osmolarity of potassium chloride in the suspending medium, confirming that the blowfly mitochondrion behaves as an osmometer. 4. Light-scattering studies showed that both respiratory substrate and a permeant anion such as phosphate or acetate are required for rapid and massive entry of K(+), which occurs in an electrophoretic process rather than in exchange for H(+). The increase in permeability to K(+) and other cations is probably the result of a large increase in the exposed area of inner membrane surface in these mitochondria, with no intrinsic increase in the permeability per unit area. 5. No increase in permeability to K(+) and other cations occurs during phosphorylation of ADP in state 3 respiration.


Suppression of melanotroph carcinogenesis leads to accelerated progression of pituitary anterior lobe tumors and medullary thyroid carcinomas in Rb+/- mice.
Mice with a single copy of the retinoblastoma gene (Rb(+/-)) develop a syndrome of multiple neuroendocrine neoplasia. They usually succumb to fast-growing, Rb-deficient melanotroph tumors of the pituitary intermediate lobe, which are extremely rare in humans. Thus, full assessment of Rb role in other, more relevant to human pathology, neoplasms is complicated. To prevent melanotroph neoplasia while preserving spontaneous carcinogenesis in other types of cells, we have prepared transgenic mice in which 770-bp fragment of pro-opiomelanocortin promoter directs expression of the human RB gene to melanotrophs (TgPOMC-RB). In three independent lines, transgenic mice crossed to Rb(+/-) background are devoid of melanotroph tumors but develop the usual spectrum of other neoplasms. Interestingly, abrogation of melanotroph carcinogenesis results in accelerated progression of pituitary anterior lobe tumors and medullary thyroid carcinomas. A combination of immunologic tests, cell culture studies, and tumorigenicity assays indicates that alpha-melanocyte-stimulating hormone, which is overproduced by melanotroph tumors, attenuates neoplastic progression by decreasing cell proliferation and inducing apoptosis. Taken together, we show that cell lineage-specific complementation of Rb function can be successfully used for refining available models of stochastic carcinogenesis and identify alpha-melanocyte-stimulating hormone as a potential attenuating factor during progression of neuroendocrine neoplasms.


Parents' Support for School-Entry Requirements for Human Papillomavirus Vaccination: A National Study.
The number of states proposing school-entry requirements for human papillomavirus (HPV) vaccination has increased over the last decade. However, data are currently limited regarding parents' support of such laws. We sought to obtain the first national estimates of parents' support of HPV vaccination school-entry requirements. A national sample of 1,501 parents of 11- to 17-year-old children completed a web-based survey between November 2014 and January 2015. Analyses used multivariable logistic regression to assess correlates of support for school-entry requirements for HPV vaccination. Overall, 21% of parents agreed that laws requiring HPV vaccination for school attendance "are a good idea," and 54% disagreed. If school-entry requirements included opt-out provisions, agreement increased to 57%, and only 21% disagreed. Parents more often agreed with requirements without opt-out provisions if they were Hispanic [OR = 1.53; 95% confidence interval (CI), 1.05-2.22], believed HPV vaccine was as or more important than other adolescent vaccines (OR = 2.76; 95% CI, 1.98-3.83), or believed HPV vaccine was effective for preventing cervical cancer (OR = 2.55; 95% CI, 1.93-3.37). Parents less often agreed if they resided in Midwest states or believed that HPV vaccine was being pushed to make money for drug companies (both P < 0.05). Opt-out provisions almost tripled parents' support for HPV vaccine school-entry requirements. Our findings suggest that race/ethnicity, attitudes about HPV vaccine, and region of residence may influence support for requirements without opt-out provisions. Opt-out provisions greatly increase parent support of school-entry requirements for HPV vaccination but may make them ineffective. Cancer Epidemiol Biomarkers Prev; 25(9); 1317-25. ©2016 AACR.


Androgen deprivation therapy for prostate cancer: recommendations to improve patient and partner quality of life.
Because of improved prostate cancer detection, more patients begin androgen deprivation therapy (ADT) earlier and remain on it longer than before. Patients now may be androgen deprived for over a decade, even when they are otherwise free of cancer symptoms. An ADT Survivorship Working Group was formed to develop and evaluate interventions to limit the physiological and emotional trauma patients and their partners experience from this treatment. The multidisciplinary Working Group met for 2 days to define the challenges couples face when patients commence ADT. A writing sub-group was formed. It compiled the meeting's proceedings, reviewed the literature and, in consultation with the other members of the working group, wrote the manuscript. Expert opinion of the side effects of ADT that affect the quality of life (QOL) of patients and their partners and the recommendations for managing ADT to optimize QOL were based on the best available literature, clinical experience, and widespread internal discussions among Working Group members. Side effects identified as particularly challenging include: (i) body feminization; (ii) changes in sexual performance; (iii) relationship changes; (iv) cognitive and affective symptoms; and (v) fatigue, sleep disturbance, and depression. Recommendations for managing ADT include providing information about ADT side effects before administration of ADT, and, where appropriate, providing referrals for psychosocial support. Sexual rehabilitation principles for persons with chronic illness may prove useful. Psychological interventions for sexual sequelae need to be offered and individualized to patients, regardless of their age or partnership. Support should also be offered to partners. Our hope is that this plan will serve as a guide for optimizing how ADT is carried out and improve the lives of androgen-deprived men and their intimate partners.


The quality of methods reporting in parasitology experiments.
There is a growing concern both inside and outside the scientific community over the lack of reproducibility of experiments. The depth and detail of reported methods are critical to the reproducibility of findings, but also for making it possible to compare and integrate data from different studies. In this study, we evaluated in detail the methods reporting in a comprehensive set of trypanosomiasis experiments that should enable valid reproduction, integration and comparison of research findings. We evaluated a subset of other parasitic (Leishmania, Toxoplasma, Plasmodium, Trichuris and Schistosoma) and non-parasitic (Mycobacterium) experimental infections in order to compare the quality of method reporting more generally. A systematic review using PubMed (2000-2012) of all publications describing gene expression in cells and animals infected with Trypanosoma spp was undertaken based on PRISMA guidelines; 23 papers were identified and included. We defined a checklist of essential parameters that should be reported and have scored the number of those parameters that are reported for each publication. Bibliometric parameters (impact factor, citations and h-index) were used to look for association between Journal and Author status and the quality of method reporting. Trichuriasis experiments achieved the highest scores and included the only paper to score 100% in all criteria. The mean of scores achieved by Trypanosoma articles through the checklist was 65.5% (range 32-90%). Bibliometric parameters were not correlated with the quality of method reporting (Spearman's rank correlation coefficient <-0.5; p>0.05). Our results indicate that the quality of methods reporting in experimental parasitology is a cause for concern and it has not improved over time, despite there being evidence that most of the assessed parameters do influence the results. We propose that our set of parameters be used as guidelines to improve the quality of the reporting of experimental infection models as a pre-requisite for integrating and comparing sets of data.


Robotic Mirror Therapy System for Functional Recovery of Hemiplegic Arms.
Mirror therapy has been performed as effective occupational therapy in a clinical setting for functional recovery of a hemiplegic arm after stroke. It is conducted by eliciting an illusion through use of a mirror as if the hemiplegic arm is moving in real-time while moving the healthy arm. It can facilitate brain neuroplasticity through activation of the sensorimotor cortex. However, conventional mirror therapy has a critical limitation in that the hemiplegic arm is not actually moving. Thus, we developed a real-time 2-axis mirror robot system as a simple add-on module for conventional mirror therapy using a closed feedback mechanism, which enables real-time movement of the hemiplegic arm. We used 3 Attitude and Heading Reference System sensors, 2 brushless DC motors for elbow and wrist joints, and exoskeletal frames. In a feasibility study on 6 healthy subjects, robotic mirror therapy was safe and feasible. We further selected tasks useful for activities of daily living training through feedback from rehabilitation doctors. A chronic stroke patient showed improvement in the Fugl-Meyer assessment scale and elbow flexor spasticity after a 2-week application of the mirror robot system. Robotic mirror therapy may enhance proprioceptive input to the sensory cortex, which is considered to be important in neuroplasticity and functional recovery of hemiplegic arms. The mirror robot system presented herein can be easily developed and utilized effectively to advance occupational therapy.


DNA-PKcs controls calcineurin mediated IL-2 production in T lymphocytes.
Loss of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) activity in mammals results in severe combined immuno-deficiency (SCID). This SCID phenotype has been postulated to be due solely to the function of DNA-PKcs in V(D)J recombination, a process critical for lymphocyte maturation. However; we show that DNA-PKcs is required for IL-2 production via regulation of the calcineurin signaling pathway. Reducing DNA-PKcs activity in activated T cells either by shRNA or an inhibitor significantly reduced IL-2 production by blocking calcineurin activity and the translocation of NFAT into the nucleus. Additionally, we show that DNA-PKcs exerts its effect on calcineurin by altering the expression of the endogenous calcineurin inhibitor Cabin1 through activation of the kinase CHK2, a known Cabin1 regulator. The discovery of DNA-PKcs as a potent regulator of IL-2 production will drive continued investigation of small molecule inhibition of this enzyme within the clinic.


The intimate relationship of gonadotropin-releasing hormone neurons with the polysialylated neural cell adhesion molecule revisited across development and adult plasticity.
The neurohormone gonadotropin-releasing hormone (GnRH) is critical for all the aspects of reproductive life in vertebrates. GnRH is secreted by a small number of neurons dispersed within the preoptic-hypothalamic region. These neurons are derived from the embryonic olfactory pit. They then migrate along olfactory, vomeronasal and terminal nerves to their final destination. Classical approaches to study the regulation of GnRH secretion during the reproductive cycle have focused on the various neuronal inputs on GnRH neurons and their regulation by ovarian steroids. However, it is well known that steroids will change the microenvironment of neuronal networks and can induce plasticity and functional changes. In this review, we will focus on the intimate relationship of developing and adult GnRH neurons with the polysialylated form of neural cell adhesion molecule (PSA-NCAM), a major molecular actor in the morphogenesis and adult plasticity of the nervous system. We will first recapitulate the spatiotemporal relationship between PSA-NCAM and migrating GnRH neurons during embryogenesis of various vertebrate species and discuss its importance for GnRH neuron development as shown by various loss of function studies. In the adult, we will review the relationships between PSA-NCAM and GnRH neurons across various physiological states, and open the discussion to the use of new model systems that can help to unravel the function and mechanism of action of PSA-NCAM on GnRH neuronal network activity and GnRH release.


Aortic Insufficiency Causes Symptomatic Heart Failure during Left Ventricular Assist Device Support.
De novo aortic insufficiency is often documented during long-term left ventricular assist device (LVAD) support, despite the absence of aortic insufficiency at the time of LVAD implantation. However, whether aortic insufficiency affects long-term mortality and symptomatic heart failure in LVAD-supported patients remains controversial. We aimed to examine whether aortic insufficiency development influenced mortality and symptomatic heart failure following LVAD implantation. Fifty-three patients who underwent durable LVAD implantation between January 1, 2008 and April 31, 2017 were retrospectively examined in a single center institute. After discharge, we performed the echocardiographic examination in accordance with the Japanese registry for the mechanically assisted circulatory support protocol. Aortic insufficiency was graded on an interval scale (severe = 4, moderate = 3, mild = 2, trivial or none = 1). Kaplan-Meier estimates for long-term mortality at the follow-up were generated. We used a logistic regression model to identify risk factors for symptomatic heart failure. The overall median duration of LVAD support was 856.3 ± 430.8 days (range, 12-1,744 days). We did not observe a significant difference in long-term mortality in patients with aortic insufficiency ≥ 3 grade compared with patients with aortic insufficiency < 3 grade (P = 0.767; log-rank). Aortic insufficiency was associated with an increased risk for heart failure event after discharge (odds ratio, 4.12; confidence interval, 1.48-16.93; P = 0.005). Aortic insufficiency was an independent risk factor for symptomatic heart failure and was not associated with long-term mortality. Aortic insufficiency progression was associated with symptomatic heart failure.


Activin Receptor-Like Kinase Receptors ALK5 and ALK1 Are Both Required for TGFβ-Induced Chondrogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells.
Bone marrow-derived mesenchymal stem cells (BMSCs) are promising for cartilage regeneration because BMSCs can differentiate into cartilage tissue-producing chondrocytes. Transforming Growth Factor β (TGFβ) is crucial for inducing chondrogenic differentiation of BMSCs and is known to signal via Activin receptor-Like Kinase (ALK) receptors ALK5 and ALK1. Since the specific role of these two TGFβ receptors in chondrogenesis is unknown, we investigated whether ALK5 and ALK1 are expressed in BMSCs and whether both receptors are required for chondrogenic differentiation of BMSCs. ALK5 and ALK1 gene expression in human BMSCs was determined with RT-qPCR. To induce chondrogenesis, human BMSCs were pellet-cultured in serum-free chondrogenic medium containing TGFβ1. Chondrogenesis was evaluated by aggrecan and collagen type IIα1 RT-qPCR analysis, and histological stainings of proteoglycans and collagen type II. To overexpress constitutively active (ca) receptors, BMSCs were transduced either with caALK5 or caALK1. Expression of ALK5 and ALK1 was downregulated by transducing BMSCs with shRNA against ALK5 or ALK1. ALK5 and ALK1 were expressed in in vitro-expanded as well as in pellet-cultured BMSCs from five donors, but mRNA levels of both TGFβ receptors did not clearly associate with chondrogenic induction. TGFβ increased ALK5 and decreased ALK1 gene expression in chondrogenically differentiating BMSC pellets. Neither caALK5 nor caALK1 overexpression induced cartilage matrix formation as efficient as that induced by TGFβ. Moreover, short hairpin-mediated downregulation of either ALK5 or ALK1 resulted in a strong inhibition of TGFβ-induced chondrogenesis. ALK5 as well as ALK1 are required for TGFβ-induced chondrogenic differentiation of BMSCs, and TGFβ not only directly induces chondrogenesis, but also modulates ALK5 and ALK1 receptor signaling in BMSCs. These results imply that optimizing cartilage formation by mesenchymal stem cells will depend on activation of both receptors.


Smartphone-based remote assessment of upper extremity function for multiple sclerosis using the Draw a Shape Test.
Smartphone devices may enable out-of-clinic assessments in chronic neurological diseases. We describe the Draw a Shape (DaS) Test, a smartphone-based and remotely administered test of Upper Extremity (UE) function developed for people with multiple sclerosis (PwMS). This work introduces DaS-related features that characterise UE function and impairment, and aims to demonstrate how multivariate modelling of these metrics can reliably predict the 9-Hole Peg Test (9HPT), a clinician-administered UE assessment in PwMS. The DaS Test instructed PwMS and healthy controls (HC) to trace predefined shapes on a smartphone screen. A total of 93 subjects (HC, n = 22; PwMS, n = 71) contributed both dominant and non-dominant handed DaS tests. PwMS subjects were characterised as those with normal (nPwMS, n = 50) and abnormal UE function (aPwMS, n = 21) with respect to their average 9HPT time (≤ or > 22.7 (s), respectively). L 1-regularization techniques, combined with linear least squares (OLS, IRLS), or non-linear support vector (SVR) or random forest (RFR) regression were investigated as functions to map relevant DaS features to 9HPT times. It was observed that average non-dominant handed 9HPT times were more accurately predicted by DaS features (r 2 = 0.41, [Formula: see text] 0.05; MAE: 2.08 ± 0.34 (s)) than average dominant handed 9HPTs (r 2 = 0.39, [Formula: see text] 0.05; MAE: 2.32 ± 0.43 (s)), using simple linear IRLS ([Formula: see text] 0.01). Moreover, it was found that the Mean absolute error (MAE) in predicted 9HPTs was comparable to the variability of actual 9HPT times within HC, nPwMS and aPwMS groups respectively. The 9HPT however exhibited large heteroscedasticity resulting in less stable predictions of longer 9HPT times. This study demonstrates the potential of the smartphone-based DaS Test to reliably predict 9HPT times and remotely monitor UE function in PwMS.


[Effect of acetazolamide on sodium and potassium absorption in the sheep forestomach].
I.v. Acetazolamide administration suppressed the sodium and potassium absorption up to 90 and 100%, resp., in the sheep reticulo-rumen. Absorption of the short chain fatty acids was not affected. The findings suggest a sodium/potassium exchange processes occurring in the reticulo-rumen epithelium in ruminants.


Does the impact of metabolic syndrome on cardiovascular events vary by using different definitions?
Metabolic Syndrome (MetS) is a complex disorder which increases the risk of chronic diseases, including cardiovascular diseases and diabetes mellitus. As a result of modern lifestyles, the prevalence of MetS has been rising globally. This study aims to investigate whether overall prevalence of MetS varies when using different definitions of MetS and to identify the best and most predictive definition of the MetS for cardiovascular disease (CVD) events over 10 years in a cohort of an Iranian population. Adults aged ≥ 35 years from urban and rural regions in central Iran were selected at baseline and followed up for more than 10 years. Data on socio-demographic characteristics, anthropometry, blood pressure and smoking status were collected at baseline. In addition, various biochemical indices were assessed. MetS was defined based on five available definitions, and cardiovascular events during 10 years follow up were confirmed by an expert group. The hazard ratios were calculated by the Cox proportional hazards model. The highest prevalence of MetS was observed by using AHA-NHBI definition (36.9%), followed by JIS definition (31.2%). On the other hand, EGIR (8.8%) provided the lowest prevalence. The risk of developing CVD, irrespective of definitions, was approximately two fold higher in the presence of MetS. After controlling for possible confounders, AHA-NHBI definition was found to be the best predictor of CVD. This study demonstrated a great variability in the prevalence of MetS among Iranian adults when using different definitions of MetS. CVD risk was significantly higher in MetS participants, as well as in participants with any risk factors of MetS; however, the AHA-NHBI definition was found to be the best predictor of CVD. Thus protective measures, including lifestyle modifications, plus control of individual risk factors is necessary to prevent cardiovascular events.


Novel sorbent shows promising financial results on P recovery from sludge water.
For several decades, researchers have been struggling to obtain minimum phosphorus (P) capture costs to meet the parameters for discharging wastewater into the watercourse. Findings from ongoing practices suggest that the Modified University of Cape Town process is currently the cheapest P capture method in the USA, whereas struvite precipitation seems to be the most cost effective method in the rest of the developed world. P sorption via biochars is becoming widespread in developing countries because this technique allows for the turning of voluminous biowaste into fertilizer with soil improving properties. Nevertheless, the reliability of this technology fluctuates throughout the year according to biowaste characteristics. For the first time, it has been proposed to use broken cellulose casings, which are readily available in increasing quantities worldwide. The sorbent obtained was subsequently activated by calcium chloride (CaCl2), whose cost is irrelevant as it would be used for agronomical purposes anyway. Pilot scale experiments show that this novel sorbent is capable of capturing 31.8 kg P t-1 from sludge water that contains 52.5 mg of extractable P L-1. More importantly, it was reported that the novel sorbent captures P, mostly in calcium phosphates (CaP) forms (191.5 g CaP t-1), which are the most valuable for plant nutrition. Enough evidence was obtained to claim that the ongoing technological race to meet the P discharge standards at the lowest cost possible should also reflect the agronomic value of P to plant nutrition to increase its competitiveness.


Antipyrine and doxycycline pharmacokinetics in patients with thyroid disorders.
Pathological conditions are known to affect pharmacokinetics of many drugs. Antipyrine half-life is used as a marker of liver microsomal enzyme function. Antipyrine pharmacokinetics, therefore, was investigated in 23 thyrotoxic and 11 euthyroid goitre patients. Of these, 11 thyrotoxic and 9 euthyroid goitre patients also participated in doxycycline bioavailability studies. In thyrotoxic patients, antipyrine half-life and AUCo infinity and doxycycline Cpmax and AUCo infinity were found to be reduced as compared to those of healthy euthyroid normal subjects. Following treatment of thyrotoxicosis, the antipyrine half-life and AUCo infinity returned to normal. Doxycycline AUCo infinity returned to near normal range but Cpmax did not.


Non contrast-enhanced MRA versus ultrasound blood vessel assessment to determine the choice of hemodialysis vascular access.
The aim of this work was to establish the relationship between traditional blood vessel mapping for vascular access (VA) creation by B-mode ultrasound (US) and novel non contrast-enhanced magnetic resonance angiography (NCE-MRA), and to study the potential influence of the diameter assessment technique on the choice of hemodialysis vascular access. A total of 27 end-stage renal-disease patients were included. They received routine US and a NCE-MRA examination of the upper extremity. Diameters were measured manually on US and semi-automatically on NCE-MRA. These measurements were statistically compared for the arteries and veins and for each measurement location. Furthermore, sensitivity and specificity of both modalities to predict VA location was investigated by comparison with an experienced surgeon. This analysis gave insight into the potential influence of vessel mapping modality on decision-making. Comparison of NCE-MRA with US for the arteries and veins, demonstrated a bias of 9% (limits -33%-78%) and 38% (limits -36%-198%), respectively. Statistically significant differences between the modalities on the individual locations were mainly found for the venous locations. The sensitivity and specificity for US to predict VA location was 1.0 and 0.74, respectively, while for NCE-MRA this was 0.88 and 0.39, respectively. The results obtained indicate that extreme caution should be exercised when replacing one diameter measurement modality with the other. A further need exists to improve both vessel mapping protocols to obtain a geometric description of the upper extremity vasculature regardless of acquisition modality.


Colonization, extinction, and phylogeographic patterning in a freshwater crustacean.
Phylogeographic analyses have revealed the importance of Pleistocene vicariance events in shaping the distribution of genetic diversity in freshwater fishes. However, few studies have examined the patterning of variation in freshwater organisms with differing dispersal syndromes and life histories. The present investigation addresses this gap, examining the phylogeography of Sida crystallina, a species whose production of diapausing eggs capable of passive dispersal was thought to constrain its regional genetic differentiation. By contrast, the present analysis has revealed deep allozyme and cytochrome oxidase I mitochondrial DNA divergence between populations from North America and Europe. Moreover, North American populations are separated into four allopatric assemblages, whose distribution suggests their derivation from different Pleistocene refugia. These lineages show higher haplotype diversity and deeper sequence divergence than those of any fish from temperate North America. Its distinctive life history traits have evidently sheltered lineages of Sida from extinction, contributing to a remarkably comprehensive and high resolution phylogeographic record.


Embryonic cord transplants in peripheral nerve restore skeletal muscle function.
The rapid atrophy of skeletal muscle after denervation severely compromises efforts to restore muscle function. We have transplanted embryonic day 14-15 (E14-E15) ventral spinal cord cells into adult Fischer rat tibial nerve stump to provide neurons for reinnervation. Our aim was to evaluate medial gastrocnemius reinnervation physiologically because this transplant strategy will only be effective if the reinnervated muscle contracts, generates sufficient force to induce joint movement, and is fatigue resistant enough to shorten repeatedly. Twelve weeks posttransplantation, brief duration electrical stimuli applied to the transplants induced medial gastrocnemius contractions that were strong enough to produce ankle movement in 4 of 12 rats (33%). The force of these four "low-threshold" reinnervated muscles and control muscles declined only gradually during five hours of intermittent, supramaximal stimulation and without depression of EMG potential area, which is strong evidence of functional neuromuscular junctions and fatigue resistant muscles. Sectioning of the medial gastrocnemius nerves confirmed that these contractions were innervation dependent. Weakness in low-threshold reinnervated muscles (8% control force) related to incomplete reinnervation, reductions in muscle fiber size, specific tension, and/or the presence of nonfunctional neuromuscular junctions. Muscle reinnervation achieved using this novel transplantation strategy may salvage completely denervated muscle and may provide the potential to evoke limb movement when injury or disease precludes or delays peripheral axon regeneration.


Iterative solution of dense linear systems arising from the electrostatic integral equation in MEG.
We study the iterative solution of dense linear systems that arise from boundary element discretizations of the electrostatic integral equation in magnetoencephalography (MEG). We show that modern iterative methods can be used to decrease the total computation time by avoiding the time-consuming computation of the LU decomposition of the coefficient matrix. More importantly, the modern iterative methods make it possible to avoid the explicit formation of the coefficient matrix which is needed when a large number of unknowns are used. To study the convergence of iterative solvers we examine the eigenvalue distributions of the coefficient matrices. For the sphere we show how the eigenvalues of the integral operator are approximated by the eigenvalues of the coefficient matrix when the collocation and Galerkin methods are used as discretization methods. The collocation method approximates the eigenvalues of the integral operator directly. The Galerkin method produces a coefficient matrix that needs to be preconditioned in order to maintain optimal convergence speed. With the ILU(0) preconditioner iterative methods converge fast and independent of the number of discretization points for both the collocation and Galerkin approaches. The preconditioner has no significant effect on the total computational time.


Born blonde: a recessive loss-of-function mutation in the melanocortin 1 receptor is associated with cream coat coloration in Antarctic fur seals.
Although the genetic basis of color variation has been extensively studied in humans and domestic animals, the genetic polymorphisms responsible for different color morphs remain to be elucidated in many wild vertebrate species. For example, hypopigmentation has been observed in numerous marine mammal species but the underlying mutations have not been identified. A particularly compelling candidate gene for explaining color polymorphism is the melanocortin 1 receptor (MC1R), which plays a key role in the regulation of pigment production. We therefore used Antarctic fur seals (Arctocephalus gazella) as a highly tractable marine mammal system with which to test for an association between nucleotide variation at the MC1R and melanin-based coat color phenotypes. By sequencing 70 wild-type individuals with dark-colored coats and 26 hypopigmented individuals with cream-colored coats, we identified a nonsynonymous mutation that results in the substitution of serine with phenylalanine at an evolutionarily highly conserved structural domain. All of the hypopigmented individuals were homozygous for the allele coding for phenylalanine, consistent with a recessive loss-of-function allele. In order to test for cryptic population structure, which can generate artefactual associations, and to evaluate whether homozygosity at the MC1R could be indicative of low genome-wide heterozygosity, we also genotyped all of the individuals at 50 polymorphic microsatellite loci. We were unable to detect any population structure and also found that wild-type and hypopigmented individuals did not differ significantly in their standardized multilocus heterozygosity. Such a lack of association implies that hypopigmented individuals are unlikely to suffer disproportionately from inbreeding depression, and hence, we have no reason to believe that they are at a selective disadvantage in the wider population.


Corynomycolic acid-containing glycolipids signal through the pattern recognition receptor Mincle.
An enantioselective synthesis of (+)-corynomycolic acid, and its elaboration to esters of trehalose, glucose and glycerol, is described. Trehalose dicorynomycolate and trehalose monocorynomycolate activate human and mouse Mincle as effectively as trehalose dicorynomycolate (cord factor). Glucose monomycolate is revealed to be a potent activator of both mouse and human Mincle. Glycerol monocorynomycolate signals through human Mincle, with the activity predominantly residing in the 2'S-isomer.


Release of folic acid in mesoporous NFM-1 silica.
Mesoporous NFM-1 silica with folic acid as template was prepared taking advantage of the supramolecular self-assembly of pterin groups and their abilities to form hexagonal liquid crystal phases. NFM-1 materials with the varied morphologies such as fiber, chiral twisting long-rod, gyroid, and amorphous particles were prepared by varying the amount of the co-structure directing agent, stirring speed and changing pH value of the synthesis. The release kinetics of NFM-1 samples with different morphologies were studied in phosphate buffer with pH = 7.4 in 37 degrees C under stirring. All the release kinetic curves are fitted by the power law and Higuchi equations. The fitting of the power law equation was separately done as for the released amount up to 60% or 100%. The materials show slow, long-term and sustained release of folic acid from mesoporous NFM-1 silica, which establishes a new foundation for the potential application in drug delivery and bioimaging.


Systematic Mutation and Unnatural Base Pair Incorporation Improves Riboswitch-Based Biosensor Response Time.
Engineered RNAs have applications in diverse fields from biomedical to environmental. In many cases, the folding of the RNA is critical to its function. Here we describe a strategy to improve the response time of a riboswitch-based fluorescent biosensor. Systematic mutagenesis was performed to either make transpose or transition base pair mutants or introduce orthogonal base pairs. Both natural and unnatural base pair mutants were found to improve the biosensor response time without compromising fold turn-on or ligand affinity. These strategies can be transferred to improve the performance of other RNA-based tools.


Expression of IL-2R, IL-4R, IL-6R on peripheral blood lymphocytes in systemic lupus erythematosus and correlation with disease activity: a prospective study.
To study the expression of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R) and interleukin-6 receptor (IL-6R) on peripheral blood lymphocytes (PBL) in patients with systemic lupus erythematosus (SLE); to correlate the level of expression of these receptors with SLE disease activity. Peripheral blood lymphocytes were studied by a high sensitivity flow cytometry technique using monoclonal antibodies directed against CD25 (IL-2R alpha chain), CD122 (IL-2R beta chain), CD124 (IL-4R), and CD126 (IL-6R). SLE disease activity was scored using the SLE Disease Activity Index, C3 and C4 concentrations, anti-dsDNA level, and absolute lymphocyte count. Compared with normal controls, PBL from patients with SLE had a higher percentage of CD25+ cells (median 20.8% v 16.5%) and a lower percentage of CD122+ cells (median 13.1% v 22.4%). The difference in CD122+ cells was greater in the CD122weak population than the CD122strong (natural killer cell) population. The percentages of CD124+ and CD126+ PBLs in patients with SLE and controls were similar. On CD25+ cells, the relative antigenic level of the IL-2R alpha chain was significantly higher in patients with SLE (median 2.01 v 1.81). The relative antigenic levels of CD122+, CD124+ and CD126+ cells were similar in patients and controls. Neither the percentages nor the relative antigenic levels of all of these cytokine receptors were correlated with any of the parameters of disease activity. Lymphocyte activation in patients with SLE was evident from the increase in CD25 expression on PBL, with a reciprocal decrease in CD122 expression. As the expression of IL-2R, IL-4R, IL-6R did not correlate with disease activity, it seems that these cytokine/receptor systems do not play a direct role in disease activation in SLE.


Carfilzomib for relapsed and refractory multiple myeloma.
Although the prognosis of multiple myeloma (MM) patients has dramatically improved during recent years, virtually all patients eventually develop relapsed refractory disease. Several new therapeutics have been developed in the last few years, including carfilzomib, a second-generation proteasome inhibitor (PI) that has been approved by the US Food and Drug Administration (FDA) in the setting of relapsed and/or refractory MM, as a single agent with or without dexamethasone, and in combination with lenalidomide in 2012 and 2015, respectively. Other promising combinations with carfilzomib are being investigated. Carfilzomib has shown superiority over the first-generation PI bortezomib on both efficacy and toxicity. In particular, profoundly lower incidence in polyneuropathy compared to bortezomib has been described. However, carfilzomib has a different toxicity profile, with more cardiovascular adverse events. Therefore, caution should be taken with the use of carfilzomib for elderly and cardiovascularly compromised patients. The once-weekly administration of carfilzomib, recently approved by the FDA in combination with dexamethasone, will lead to a lower burden for the patient and caregivers compared to the twice-weekly schemes that were routinely used until recently. This review has a focus on clinical trial data that has led to drug approval, as well as new promising combination studies, and provides advice for treating physicians who are now prescribing this drug to patients.


Peptidylprolyl cis/trans isomerases (immunophilins): biological diversity--targets--functions.
Information recovered from genome sequencing projects, multiple sequence alignments, structural analyses of PPIase and published records were used in deciphering the biological diversity, functions and targets of four groups of proteins encoded by dissimilar sets of sequences whose spatial representations exhibit peptidylprolyl cis/trans isomerase activity (PPIase). In the human genome there are encoded fifteen proteins whose segments have significant homology with the sequence of 12 kDa protein which is the target of the potent immunosuppressive macrolides FK506 or rapamycin. The 12 kDa archetype of the FK506-binding protein (FKBP), known as FKBP-12a, is an abundant intracellular protein whereas other FKBPs possessing from one to four FK506-like binding domains (FKBDs) have nominal masses varying from 13 to 135 kDa. The human genome contains at least sixteen genes encoding proteins comprising one cyclosporin-A (CsA) binding domain (CLD) called cyclophilins whose nominal masses vary from 17 to 324 kDa and multiple coding segments for small cyclophilins (17-19 kDa) whose transcription levels and functions remain unknown. The third group of PPIases encoded in the genome comprises two proteins (hPin1 and hParv14) where hPin1 is an important PPIase for cell cycle. The A. thaliana, C. elegans, D. melanogaster and S. cerevisiae genomes encode a less diverse spectrum of PPIases whereas the prokaryotic genomes contain from none to three cyclophilins, from none to four genes encoding FKBPs, one distant homologue of the Pin1 protein named parvulin and the fourth group of PPIases known as trigger factors. PPIases are discretely distributed to different cellular compartments and interact with a number of targets that control a range of cellular processes. Analyses of the sequence alignments of the two groups of PPIases, namely cyclophilins and FKBPs from diverse phyla, show that in each group their sequences diverge but the amino acid residues which form the PPIase activity site and macrolide binding cavity remain well conserved in the majority of them which suggests that the spatial structures and functions of each group of PPIases remain conserved.


Failure of gastroplasty pouch and stoma size to correlate with postoperative weight loss.
At the University of Alberta hospitals patients who underwent gastroplasty--horizontal, vertical with multistranded nylon banding of the stomas and vertical banded with Teflon reinforced stomas--have been followed up for over 2 years. Pouch volume ranged from 25 to 30 mL. Pouch volume and stoma size were measured radiologically 2 years after operation and correlated with the percentage excess weight loss. Any patient failing to lose at least 40% of the excess weight was considered a failure. There was no consistent correlation between pouch or stoma size and percentage excess weight loss for any of the three procedures studied. The failure rate for patients included in the study group was 33.3% for horizontal, 32.4% for vertical and 32.8% for vertical banded gastroplasty. The probable reason for the lack of correlation is that some patients change their diets so as to subvert the restriction imposed by the gastroplasty.


Identification of a novel class of endoperoxides from arachidonate autoxidation.
Free radical-initiated lipid autoxidation in low density lipoprotein (LDL) has been implicated in the pathogenesis of atherosclerosis. Oxidation of the lipid components of LDL leads to a complex mixture of hydroperoxides, bicyclic endoperoxides, monocyclic peroxides, and serial cyclic peroxides. The oxidation compounds and/or their decomposition products can modify protein components, which may lead to various diseases. A novel class of peroxides (termed dioxolane-isoprostanes) having a bicyclic endoperoxide moiety characteristic of the isoprostanes and a dioxolane peroxide functionality in the same molecule was identified in the product mixture formed from in vitro autoxidation of cholesteryl arachidonate. The same products are also detected in in vitro oxidized LDL. Various mass spectrometric techniques have been applied to characterize these new peroxides. The structure of these compounds has also been confirmed by independent synthesis. We reason, based on the free radical mechanism of the transformation, that only the 12- and 8-peroxyl radicals (those leading to 12-HPETE and 8-HPETE) of arachidonate can form these new peroxides. We also suggest that the formation of these peroxides provides a rationale to explain the fact that 5- and 15-series isoprostanes are formed in preference to 8- and 12-series. Furthermore, series of other isoprostanes, such as dioxolane A(2), D(2), E(2), etc., can be derived from the dioxolane-isoprostane peroxides. These findings offer further insights into the oxidation products of arachidonate and the opportunity to study their potential biological relevance.


Site-dependent expression of vascular endothelial growth factor, angiogenesis and proliferation in human gastric carcinoma.
To determine the role of site of tumor implantation on tumor angiogenesis, we implanted gastric cancer cells in the orthotopic (stomach) and ectopic (subcutaneous) locations in nude mice. Tumors in the stomach demonstrated greater vascularization, higher levels of vascular endothelial growth factor (VEGF) expression, and greater proliferation compared with tumors in the subcutaneous tissues. These data suggest that the relationships among the expression of VEGF, vascularization, and proliferation of human gastric cancer cells are regulated by the organ microenvironment. In addition, VEGF may provide a target for anti-angiogenic therapy for gastric cancers.


14 beta-Hydroxygelsedine, a new oxindole alkaloid from Gelsemium sempervirens.
An extract of the stem of Gelsemium sempervirens has afforded a new oxindole alkaloid, 14 beta-hydroxygelsedine, whose structure and stereochemistry were deduced by spectral methods, principally high resolution 1H nmr, 2-D COSY, and decoupling experiments.


Changes in LV papillary muscle performance and myosin composition with aortic insufficiency in rats.
Aortic insufficiency was induced in rats. Left ventricular papillary muscle function was studied after 5, 12, and 40 wk and compared with the papillary muscles from sham-operated animals. The maximum unloaded velocity of shortening, Vmax, was decreased in the rats with aortic insufficiency relative to controls by 15, 20, and 34% at 5, 12, and 40 wk, respectively. The decrease in Vmax occurred concomitantly with a change in the myosin isoenzyme composition such that the V1 isoform content decreased and the V3 isoform increased. Relative to age-matched controls, the V3 content in the hearts with aortic insufficiency had increased by 80, 180, and 125% at 5, 12, and 40 wk, respectively. The decrease in Vmax in the aortic insufficiency group muscles correlated with the change in myosin isozyme composition and could not be explained by changes in collagen content. Thus aortic insufficiency induced changes in myosin isozyme content and Vmax similar to those previously observed with aortic stenosis, thus suggesting a common mechanism of myocardial adaptation to different types of mechanical overload.


Tracheobronchial Injury caused by Blunt Trauma: Case Report and Review of Literature.
Tracheobronchial injuries are rare cases requiring skillful airway management. We report a challenging case of tracheobronchial injury in a young adult who was run over by a tractor and was referred to us from a peripheral hospital with endotracheal tube in situ. He was severely hypoxaemic on initial presentation. Diagnostic work up showed high suspicion for right bronchial transection along with left lung upper lobe contusion. Due to deteriorating clinical condition of the patient and despite immediate unavailability of fibreoptic bronchoscope, patient was immediately taken up for right posterolateral thoracotomy and double lumen tube was inserted. The position of the tube was confirmed clinically. As soon as the fibreoptic bronchoscope arrived, it was again used to confirm the position of double lumen tube. Patient's clinical condition improved after repair of the injured right bronchus and he was later extubated the next day.


SARS-CoV-2 N Protein Targets TRIM25-Mediated RIG-I Activation to Suppress Innate Immunity.
A weak production of INF-β along with an exacerbated release of pro-inflammatory cytokines have been reported during infection by the novel SARS-CoV-2 virus. SARS-CoV-2 encodes several proteins able to counteract the host immune system, which is believed to be one of the most important features contributing to the viral pathogenesis and development of a severe clinical picture. Previous reports have demonstrated that SARS-CoV-2 N protein, along with some non-structural and accessory proteins, efficiently suppresses INF-β production by interacting with RIG-I, an important pattern recognition receptor (PRR) involved in the recognition of pathogen-derived molecules. In the present study, we better characterized the mechanism by which the SARS-CoV-2 N counteracts INF-β secretion and affects RIG-I signaling pathways. In detail, when the N protein was ectopically expressed, we noted a marked decrease in TRIM25-mediated RIG-I activation. The capability of the N protein to bind to, and probably mask, TRIM25 could be the consequence of its antagonistic activity. Furthermore, this interaction occurred at the SPRY domain of TRIM25, harboring the RNA-binding activity necessary for TRIM25 self-activation. Here, we describe new findings regarding the interplay between SARS-CoV-2 and the IFN system, filling some gaps for a better understanding of the molecular mechanisms affecting the innate immune response in COVID-19.


WAM: an improved algorithm for modelling antibodies on the WEB.
An improved antibody modelling algorithm has been developed which incorporates significant improvements to the earlier versions developed by Martin et al. (1989, 1991), Pedersen et al. (1992) and Rees et al. (1996) and known as AbM (Oxford Molecular). The new algorithm, WAM (for Web Antibody Modelling), has been launched as an online modelling service and is located at URL http://antibody.bath.ac.uk. Here we provide a summary only of the important features of WAM. Readers interested in further details are directed to the website, which gives extensive background information on the methods employed. A brief description of the rationale behind some of the newer methodology (specifically, the knowledge-based screens) is also given.


Using dynamic Brownian Bridge Movement Models to identify home range size and movement patterns in king cobras.
Home range estimators are a critical component for understanding animal spatial ecology. The choice of home range estimator in spatial ecology studies can significantly influence management and conservation actions, as different methods lead to vastly different interpretations of movement patterns, habitat selection, as well as home range requirements. Reptile studies in particular have struggled to reach a consensus on the appropriate home range estimators to use, and species with cryptic behavior make home range assessment difficult. We applied dynamic Brownian Bridge Movement Models (dBBMMs) to radio-telemetry data from Ophiophagus hannah, a wide-ranging snake species. We used two focal individuals at different life stages (one juvenile male and one adult male) and sought to identify whether the method would accurately represent both their home range and movement patterns. To assess the suitability of dBBMMs, we compared this novel method with traditional home range estimation methods: minimum convex polygons (MCP) and Kernel density estimators (KDE). Both KDE and MCP incorporated higher levels of Type I and Type II errors, which would lead to biases in our understanding of this species space-use and habitat selection. Although these methods identified some general spatial-temporal patterns, dBBMMs were more efficient at detecting movement corridors and accurately representing long-term shelters sites, showing an improvement over methods traditionally favored in reptile studies. The additional flexibility of the dBBMM approach in providing insight into movement patterns can help further improve conservation and management actions. Additionally, our results suggest that dBBMMs may be more widely applicable in studies that rely on VHF telemetry and not limited to studies employing GPS tags.


Addressing Safe Opioid Monitoring Practices Using an Interprofessional Approach.
The aims of the project are to increase knowledge related to opioids and improve adherence to evidence-based interventions for sedation monitoring and assessment in adult inpatients receiving opioids for pain through delivery of an interprofessional online program. Preventable opioid-induced respiratory depression is a patient safety risk in hospitals. Sedation and respiratory assessment and monitoring guidelines and tools are available to improve practice. An interprofessional team identified organization-specific drivers of opioid-induced sedation and respiratory depression using process improvement methodology, developed interventions, and evaluated clinician opioid knowledge and documentation practice. Opioid knowledge survey responses improved after the interventions. Audits showing improved appropriate documentation by RNs of the correct sedation scale were performed over 2 separate 3-month periods. Clinicians demonstrated increased knowledge about newly adopted sedation and monitoring practices targeted to prevent opioid-induced respiratory depression.


Should Zelen pre-randomised consent designs be used in some neonatal trials?
My aim is to suggest that there is a case for using a randomised consent design in some neonatal trials. As an example I use the trials of extracorporeal membrane oxygenation (ECMO) in neonates suffering pulmonary hypertension. In some trials the process of obtaining consent has the potential to harm the subject, for example, by disappointing those who end in the control group and by creating additional anxiety at times of acute illness. An example of such were the trials of extracorporeal membrane oxygenation (ECMO) in neonates suffering pulmonary hypertension. Pre-randomised consent could avoid or lessen these harms. However, a number of ethical objections are made to these research designs. They involve denial of information, using people, denial of choice, and "overselling" of allocated treatment. Furthermore, they are the wrong response; better communication might be the answer, for example. I argue that these objections are not completely persuasive. However, they are enough to suggest caution in the use of such designs.


Microparticles formulated from a family of novel silylated polysaccharides demonstrate inherent immunostimulatory properties and tunable hydrolytic degradability.
Acid-degradable polymers are well-suited for use as drug delivery vehicles because numerous physiological sites (e.g., intracellular endocytic pathway) are acidic. Here we report the synthesis of acid-sensitive silylated polysaccharides derived from either dextran or inulin with various alkyl substitutions on the silicon center: trimethylsilyl dextran (TMS-DEX), ethyldimethylsilyl dextran (EDMS-DEX), triethylsilyl dextran (TES-DEX), and trimethylsilyl inulin (TMS-IN). The silylated dextran (Silyl-DEX) and silylated inulin (Silyl-IN) polymers were fabricated into microparticles (MPs) via emulsification followed by solvent evaporation. These MPs were relatively stable at extracellular pH 7.4 and displayed a wide range of pH 2.0 and 5.0 degradation half-lives (fifteen minutes to greater than nine days) that were dependent on the extent of silylation (40 to 98%) and steric crowding on the silicon center (trimethyl to ethyldimethyl to triethyl). Silyl-DEX and Silyl-IN MPs exhibited cytocompatibility when cultured in vitro with RAW 264.7 macrophages. TES-DEX and TMS-IN MPs, composed of highly hydrophobic moieties and the parent immunostimulatory inulin, respectively, elicited substantial in vitro production of tumor necrosis factor alpha, a cytokine associated with an innate immune response. In vivo immunization with a model ovalbumin antigen encapsulated in silylated polysaccharide MPs, without a separate adjuvant, resulted in a dual humoral and cellular response that was superior to an alum-adjuvanted formulation. Overall, we present Silyl-DEX and Silyl-IN as members of the acid-degradable polymer family for potential use in subunit vaccines and other drug delivery applications.


Predicting naming latencies for action pictures: Dutch norms.
The present study provides Dutch norms for age of acquisition, familiarity, imageability, image agreement, visual complexity, word frequency, and word length (in syllables) for 124 line drawings of actions. Ratings were obtained from 117 Dutch participants. Word frequency was determined on the basis of the SUBTLEX-NL corpus (Keuleers, Brysbaert, & New, Behavior Research Methods, 42, 643-650, 2010). For 104 of the pictures, naming latencies and name agreement were determined in a separate naming experiment with 74 native speakers of Dutch. The Dutch norms closely corresponded to the norms for British English. Multiple regression analysis showed that age of acquisition, imageability, image agreement, visual complexity, and name agreement were significant predictors of naming latencies, whereas word frequency and word length were not. Combined with the results of a principal-component analysis, these findings suggest that variables influencing the processes of conceptual preparation and lexical selection affect latencies more strongly than do variables influencing word-form encoding.


Imaging cone photoreceptors in three dimensions and in time using ultrahigh resolution optical coherence tomography with adaptive optics.
Cone photoreceptors in the living human eye have recently been imaged with micron-scale resolution in all three spatial dimensions using adaptive optics optical coherence tomography. While these advances have allowed non-invasive study of the three-dimensional structure of living human cones, studies of their function and physiology are still hampered by the difficulties to monitor the same cells over time. The purpose of this study is to demonstrate the feasibility of cone monitoring using ultrahigh-resolution adaptive optics optical coherence tomography. Critical to this is incorporation of a high speed CMOS camera (125 KHz) and a novel feature-based, image registration/dewarping algorithm for reducing the deleterious effects of eye motion on volume images. Volume movies were acquired on three healthy subjects at retinal eccentricities from 0.5° to 6°. Image registration/dewarping reduced motion artifacts in the movies from 15 μm to 1.3 μm root mean square, the latter sufficient for identifying and tracking cones. Cone row-to-row spacing and outer segment lengths were consistent with that reported in the literature. Cone length analysis demonstrates that UHR-AO-OCT is sufficiently sensitive to measure real length differences between cones in the same 0.5° retinal patch, and requires no more than five measurements of OS length to achieve 95% confidence. We know of no other imaging modality that can monitor foveal or parafoveal cones over time with comparable resolution in all three dimensions.


Too shameful to remember: a 17-year-old Chinese boy with psychogenic amnesia.
A case of psychogenic amnesia in a 17-year-old Chinese boy is presented. The case is unusual in that the amnesia lasted as long as six weeks without any pseudodementia or fugue. The patient can be best understood from the abnormal sick role and the communication models of hysteria. The family was preoccupied with physical illness and relied on non-verbal rather than verbal language in communicating distress and negative emotions. Shame was the central psychodynamic aetiology; it was too shameful for the patient to talk about or withdraw himself from the predicaments in which he was trapped. Preceding the onset of amnesia he had suffered from multiple conversion symptoms for seven months. His message and predicaments not understood, he was unnecessarily investigated and treated by numerous doctors, making matters much worse. The prices paid included five hospital admissions, countless consultations at accident and emergency departments and with general practitioners, and an appendectomy for a normal appendix.


Evaluation of an Australian miscarriage support programme.
Miscarriage is a frequent and distressing event for women who often require admission to a gynaecology unit for the evacuation of the products of conception. Most women are admitted for day surgery or an overnight stay. The short time frame limits the time nursing staff have to provide support and information for women who have experienced loss through miscarriage. This article presents the qualitative evaluation of a telephone follow-up service provided as part of the Miscarriage Support Programme of Care conducted by staff of the gynaecology inpatient services in an Australian public hospital. The data from this evaluation indicate that miscarriage does have a varying degree of impact on a woman's inpatient experience and emotional recovery. The primary themes of emotional recovery, support after discharge, inpatient experience of care, and usefulness of the literature given to women before discharge, will guide the ongoing development of support and discharge planning by staff of gynaecology inpatient services for women who have experienced miscarriage.


Effect of heparin on neuroprotection against spinal cord ischemia and reperfusion in rats.
Paraplegia due to ischemia/reperfusion (I/R) injury of the spinal cord is a devastating and undesired complication of thoraco-abdominal aortic surgery. Unidentified clots cause a variety of thromboembolic events and deteriorate the severity of ischemia. We investigated the effect of the degree of anticoagulation on spinal cord I/R injury and whether heparin is protective against I/R injury beside its anticoagulant properties. Twenty-eight rats were randomly assigned to four groups (n=7 per group) as G1 (no aortic occlusion and heparin administration), G2 (45 min aortic occlusion; no heparin administration), G3 (45 min aortic occlusion; 400 IU/kg heparin to keep activated clotting time (ACT) level around 200 sec), and G4 (45 min aortic occlusion; 800 IU/kg heparin to keep ACT level around 600 sec). After neurologic evaluation at the 48th hour of reperfusion, lumbar spinal cords were removed for histopathologic evaluation and immunohistochemical staining for HSP70 (heat shock protein 70), interleukin-6 and myeloperoxidase (MPO). The Motor Deficit Index (MDI) scores were lowest in G1 group (p < 0.05) and the MDI scores of G3 and G4 were significantly lower than G2 group (p < 0.05). The neuronal degeneration in G3 was significantly lower than the other groups, respectively (p = 0.03). Histopathological evaluation showed no significant intergroup differences in terms of the degree of edema and inflammatory response. There was no statistically significant difference found among the groups in terms of HSP70 staining, IL-6 staining or the degree of MPO staining. Protection of spinal cord from I/R injury requires a multimodal management. We should not miss out the importance of adequate anticoagulation in thoraco-abdominal surgical procedures. Furthermore, the recently discovered anti-inflammatory property of glycosaminoglycans, including heparin, deserves to be investigated.


Early Deterioration and Long-Term Prognosis of Patients With Intracerebral Hemorrhage Along With Hematoma Volume More Than 20 ml: Who Needs Surgery?
Background and Purpose: The treatment of patients with intracerebral hemorrhage along with moderate hematoma and without cerebral hernia is controversial. This study aimed to explore risk factors and establish prediction models for early deterioration and poor prognosis. Methods: We screened patients from the prospective intracerebral hemorrhage (ICH) registration database (RIS-MIS-ICH, ClinicalTrials.gov Identifier: NCT03862729). The enrolled patients had no brain hernia at admission, with a hematoma volume of more than 20 ml. All patients were initially treated by conservative methods and followed up ≥ 1 year. A decline of Glasgow Coma Scale (GCS) more than 2 or conversion to surgery within 72 h after admission was defined as early deterioration. Modified Rankin Scale (mRS) ≥ 4 at 1 year after stroke was defined as poor prognosis. The independent risk factors of early deterioration and poor prognosis were determined by univariate and multivariate regression analysis. The prediction models were established based on the weight of the independent risk factors. The accuracy and value of models were tested by the receiver operating characteristic (ROC) curve. Results: After screening 632 patients with ICH, a total of 123 legal patients were included. According to statistical analysis, admission GCS (OR, 1.43; 95% CI, 1.18-1.74; P < 0.001) and hematoma volume (OR, 0.9; 95% CI, 0.84-0.97; P = 0.003) were the independent risk factors for early deterioration. Hematoma location (OR, 0.027; 95% CI, 0.004-0.17; P < 0.001) and hematoma volume (OR, 1.09; 95% CI, 1.03-1.15; P < 0.001) were the independent risk factors for poor prognosis, and island sign had a trend toward significance (OR, 0.5; 95% CI, 0.16-1.57; P = 0.051). The admission GCS and hematoma volume score were combined for an early deterioration prediction model with a score from 2 to 5. ROC curve showed an area under the curve (AUC) was 0.778 and cut-off point was 3.5. Combining the score of hematoma volume, island sign, and hematoma location, a long-term prognosis prediction model was established with a score from 2 to 6. ROC curve showed AUC was 0.792 and cutoff point was 4.5. Conclusions: The novel early deterioration and long-term prognosis prediction models are simple, objective, and accurate for patients with ICH along with a hematoma volume of more than 20 ml.


Kinesin-12 Kif15 targets kinetochore fibers through an intrinsic two-step mechanism.
Proteins that recognize and act on specific subsets of microtubules (MTs) enable the varied functions of the MT cytoskeleton. We recently discovered that Kif15 localizes exclusively to kinetochore fibers (K-fibers) or bundles of kinetochore-MTs within the mitotic spindle. It is currently speculated that the MT-associated protein TPX2 loads Kif15 onto spindle MTs, but this model has not been rigorously tested. Here, we show that Kif15 accumulates on MT bundles as a consequence of two inherent biochemical properties. First, Kif15 is self-repressed by its C terminus. Second, Kif15 harbors a nonmotor MT-binding site, enabling dimeric Kif15 to crosslink and slide MTs. Two-MT binding activates Kif15, resulting in its accumulation on and motility within MT bundles but not on individual MTs. We propose that Kif15 targets K-fibers via an intrinsic two-step mechanism involving molecular unfolding and two-MT binding. This work challenges the current model of Kif15 regulation and provides the first account of a kinesin that specifically recognizes a higher-order MT array.


[Diagnostic and prognostic significance of biochemical indicators in colorectal neoplasms].
Activities of some enzymes and content of medium-weight molecules in patients with colorectal cancer were studied in order to assess the diagnostic value of these parameters for detection of tumors in the large intestine and development of endogenous intoxication after surgery and for prediction of remote results of treatment. Increased activities of creatine phosphokinase and hexokinase is typical of tumor growth, whereas increased activities of alkaline phosphatase and gamma-glutamyl transpeptidase are observed only in metastases to the liver. The role of medium-weight molecules in the diagnosis of endogenous intoxication during the early postoperative period is shown. The content of these molecules in the sera increases 3 days before clinical manifestation of endotoxicosis, when the traditional parameters are virtually normal.


The application of biosecurity practices for preventing avian influenza in North-Eastern Italy turkey farms: An analysis of the point of view and perception of farmers.
Italian and international outbreaks of highly pathogenic avian influenza (HPAI), particularly in densely populated poultry areas (DPPAs), have increased over the past few decades. These emerging risks, which endanger both human and animal health and the entire poultry industry, can be effectively limited by biosecurity measures implemented at human-animal food chain interfaces. Some problems, however, persist in the application of these measures on the part of poultry farmers, prompting the need to explore those aspects and causes that limit their implementation. A qualitative approach was selected for the study and a semi-structured interview technique was applied to collect data among turkey farmers (n = 29) working in the north-east of Italy. The aim of this technique was to gather data on farms in order to understand the biosecurity practices adopted and the reasons for and impediments to farmer implementation, or lack thereof. This article presents and discusses the main data collected. The study revealed that farmers were familiar with the biosecurity measures necessary to contain avian influenza (AI) and other poultry diseases; personal disinfection and animal isolation practices were particularly prominent. Based on the reported procedures, managerial, economic, and psychosocial factors were among the barriers behind the failure to implement biosecurity measures. These obstacles were variously intertwined and associated with the different action settings. In particular management factors, such as lack of time to apply the rules and difficulties contingent on the farm's structural characteristics, mediate the application of biosecurity measures. In terms of communication channels, the company, particularly its technicians, proved to be the primary source of information for farmers in case of emergencies, as well as the primary source of information on the application of biosecurity measures. However, other sources of information were indicated, such as word of mouth among farmers or other non-institutional figures (relatives and acquaintances). What emerged, was the need to improve not only the biosecurity management skills, but also to implement forms of cooperation among the various key stakeholders in the poultry sector. The information presented in this pilot study needs to be discussed among competent authorities, public and company veterinarians, company technicians, and farmers. Furthermore, this information will help in participatory co-planning of risk prevention and communication strategies to implement a long-term, sustainable, effective approach to address future epidemic emergencies.


A distinction concerning vision-for-action and affordance perception.
In this paper, I offer a discussion concerning the conceptual connection between the notion of vision-for-action and the one of affordance perception. I first analyze the notion of vision-for-action. I then analyze a notion usually coupled with it: the notion of affordance perception, the main insights behind which are guiding several current neuroscientific enterprises and the related philosophical speculations. Then, I argue that we should not couple these two notions with a light heart: though these two processes can be, from a theoretical point of view, related, we should be careful in inferring the actual and effective occurrence of the latter in the presence of the former. This will be done by carrying out a conceptual analysis of the experimental evidence coming from the 'Two Visual Systems Model', which is the main reference in the literature on affordance perception and vision-for-action. My point has strong philosophical implications for our view concerning the best interpretation of how vision-for-action really works, and the specific relation it actually entertains with affordance perception.


Optimised amino acid specific weighting factors for unbound protein docking.
One of the most challenging aspects of protein-protein docking is the inclusion of flexibility into the docking procedure. We developed a postfilter where the grid-representation of proteins for docking is extended by an optimised weighting factor for each amino acid. For up to 86% of the evaluated complexes a near-native structure was within the top 5% of the ranked prediction output. The weighting factors obtained by the optimisation procedure correlate to a certain extent with the flexibility of the amino acids, their hydrophobicity and with their propensity to be in the interface. Use of the optimised amino acid specific parameters yields a strong increase of near-native structures on the first ranks of the prediction.


Transesophageal versus intracardiac atrial stimulation in assessing electrophysiologic parameters of the sinus and AV nodes and of the atrial myocardium.
Electrophysiological parameters of the sinus and AV nodes and of the atrial myocardium were assessed with both transesophageal atrial stimulation (TAS) and intracardiac atrial stimulation (ICS) in the same patient during the same study. The study group was comprised of nine men and seven women, aged 45 to 79 years, referred for the evaluation of syncope of possible arrhythmogenic origin. Twelve patients were included for analysis. Autonomic inhibition (AI) was obtained in five patients. The most striking result was the significantly longer AERP with TAS (mean 286 +/- 9 ms) than with ICS (mean 244 +/- 12 ms; p than 0.02). After AI, the AERP was even more prolonged with TAS (mean 332 +/- 20 ms) than with ICS (mean 237 +/- 8 ms; p less than 0.01). Intraatrial and AV nodal conduction times assessed at multiple paced cycle lengths were significantly shorter with TAS than with ICS. There was no difference between TAS and ICS with regard to AVERP, Wenckebach periodicity and H-V intervals. Although a tendency towards shorter sinus node recovery time (SNRT) and sinoatrial conduction time (SACT) was observed with TAS, the difference was not statistically significant. Possible mechanisms of the differences are discussed. It seemed clear that the site of origin of an atrial impulse can have definite effects upon excitability and conduction properties of atrial and AV nodal fibers. Enhanced sympathetic activity during TAS was also suggested. The electrophysiological properties inherent in the TAS technique warrant further elucidation.


[Specific for apurine-apyrimidine DNA endodeoxyribonuclease from the rat brain].
Endodeoxyribonuclease was detected in rat neocortex chromatin. The partly purified enzyme was found to influence the superhelical apurine-apyrimidine DNA 50 times as effectively as compared to the native substrate. The enzyme hydrolyzes the phosphodiester bond with the formation of 3'-OH- and 5'-phosphate terminal groups. The enzyme-hydrolyzed DNA is an effective primer for DNA-polymerase I from E. coli. It was assumed that DNAase from rat brain chromatin is an apurine-apyrimidine endonuclease II.


Baicalein Enhances the Effect of Acarbose on the Improvement of Nonalcoholic Fatty Liver Disease Associated with Prediabetes via the Inhibition of De Novo Lipogenesis.
Prediabetes is a prevalent metabolic disorder with multiple complications, including nonalcoholic fatty liver disease (NAFLD). In this study, we investigated the combinatorial effect of baicalein, a dietary flavonoid abundant in multiple edible plants, and acarbose on prediabetes-associated NAFLD. Baicalein and its metabolites inhibited de novo lipogenesis (DNL), thereby decreasing lipid accumulation and hepatokine secretion in oleic acid-induced hepatocytes. Carbohydrate restriction, which mimicked the effect of acarbose, led to comparable results. The combinatorial effect of baicalein and acarbose was further verified in prediabetic mice with NAFLD. Through the 16-week intervention, baicalein and acarbose inhibited DNL and improved glucose tolerance, oxidative stress, liver histology, and hepatokine secretion, thereby ameliorating insulin resistance and NAFLD. Our study demonstrated that baicalein enhanced the effect of acarbose on improving NAFLD and explored the underlying multitarget mechanism, laying a theoretical foundation for the development of flavonoid dietary supplements for the simultaneous improvement of NAFLD and prediabetes.


Hospitalisation with injection-related infections: Validation of diagnostic codes to monitor admission trends at a tertiary care hospital in Melbourne, Australia.
Injection-related infections (IRI) cause morbidity and mortality in people who inject drugs. Hospital administrative datasets can be used to describe hospitalisation trends, but there are no validated algorithms to identify injecting drug use and IRIs. We aimed to validate International Classification of Diseases (ICD) codes to identify admissions with IRIs and use these codes to describe IRIs within our hospital. We developed a candidate set of ICD codes to identify current injecting drug use and IRI and extracted admissions satisfying both criteria. We then used manual chart review data from 1 January 2017 to 30 April 2019 to evaluate the performance of these codes and refine our algorithm by selecting codes with a high-positive predictive value (PPV). We used the refined algorithm to describe trends and outcomes of people who inject drugs with an IRI at Alfred Hospital, Melbourne from 2008 to 2020. Current injecting drug use was best predicted by opioid-related disorders (F11), 80% (95% confidence interval [CI] 74-85%), and other stimulant-related disorders (F15), 82% (95% CI 70-90%). All PPVs were ≥67% to identify specific IRIs, and ≥84% for identifying any IRI. Using these codes over 12 years, IRIs increased from 138 to 249 per 100 000 admissions, and skin and soft tissues infections (SSTI) were the most common (797/1751, 46%). Validated ICD-based algorithms can inform passive surveillance systems. Strategies to reduce hospitalisation with IRIs should be supported by early intervention and prevention, particularly for SSTIs which may represent delayed access to care.


Competing risk of atherosclerotic risk factors for arterial and venous thrombosis in a general population: the Tromso study.
To investigate and compare the impact of traditional atherosclerotic risk factors for the risk of arterial and venous thrombosis, taking into account competing risks. In 1994-1995, 26,185 subjects were screened in the Tromsø study. Information on traditional atherosclerotic risk factors was obtained by physical examination, blood samples, and questionnaires. Subjects were followed to the first incident event of myocardial infarction (MI) or venous thromboembolism (VTE), or December 31, 2005. During a median of 10.8 years of follow-up, there were 1279 cases of incident MI and 341 VTE events. Advancing age and high body mass index were both associated with MI and VTE. Hazard ratio per decade of age was 2.34 (95% CI: 2.25-2.43) for MI and 1.87 (1.74-2.01) for VTE, and 3 kg/m(2) increase in body mass index was associated with 1.16 (1.11-1.21) and 1.20 (1.12-1.29) increased risk of MI and VTE, respectively. Blood pressure, high levels of triglycerides and total cholesterol, low HDL cholesterol, self-reported diabetes, and smoking were all associated with increased risk of MI but not associated with VTE. Our findings imply that traditional atherosclerotic risk factors, such as smoking, hypertension, dyslipidemia, and diabetes mellitus are not shared by arterial and venous thrombosis.


A chimeric cell adhesion molecule mediates homing of lymphocytes to vascularized tumors.
To facilitate tumor colonization by adoptively transferred cells of the immune system, we created a chimeric cell adhesion molecule that mediates tumor-specific homing by binding to the integrin alpha(v)beta3 on angiogenic endothelial cells. A high-affinity cell adhesion molecule for integrin alpha(v)beta3 was generated by fusing the disintegrin kistrin to the transmembrane adhesion molecule CD31/PECAM-1. This chimeric cell adhesion molecule, termed KISS31, mediates adhesion of lymphoid cells to soluble recombinant integrin alpha(v)beta3 and to endotheliomal monolayers in vitro. KISS31-expressing lymphoid cells accumulate in angiogenic tumors in two in vivo models, in B16/129 melanoma xenografts on the chick chorioallantois and in s.c. growing Lewis lung carcinoma in mice. Our data indicate that expression of KISS31 on lymphoid cells confers tumor-specific homing. This is, to our knowledge, the first example of an experimental mechanism that targets living cells to tumors by redirecting their homing pattern.


Integration of an organic photodetector onto a plastic optical fiber by means of spray coating technique.
A working prototype of integrated fiber/receiver system for optical data transmission is realized. The prototype is made by directly depositing an organic photodetector onto a plastic optical fiber. For the deposition of the organic layers, spray coating is successfully exploited. Operation over four orders of magnitude range of light intensities as well as photoresponse to pulsed stimulation are demonstrated.


Visualization of estrogen receptor transcriptional activation in zebrafish.
Estrogens regulate a diverse range of physiological processes and affect multiple tissues. Estrogen receptors (ERs) regulate transcription by binding to DNA at conserved estrogen response elements, and such elements have been used to report ER activity in cultured cells and in transgenic mice. We generated stable, transgenic zebrafish containing five consecutive elements upstream of a c-fos minimal promoter and green fluorescent protein (GFP) to visualize and quantify transcriptional activation in live larvae. Transgenic larvae show robust, dose-dependent estrogen-dependent fluorescent labeling in the liver, consistent with er gene expression, whereas ER antagonists inhibit GFP expression. The nonestrogenic steroids dexamethasone and progesterone fail to activate GFP, confirming ER selectivity. Natural and synthetic estrogens activated the transgene with varying potency, and two chemicals, genistein and bisphenol A, preferentially induce GFP expression in the heart. In adult fish, fluorescence was observed in estrogenic tissues such as the liver, ovary, pituitary gland, and brain. Individual estrogen-responsive neurons and their projections were visualized in the adult brain, and GFP-positive neurons increased in number after 17β-estradiol exposure. The transgenic estrogen-responsive zebrafish allow ER signaling to be monitored visually and serve as in vivo sentinels for detection of estrogenic compounds.


Reforming the Portuguese mental health system: an incentive-based approach.
To promote an effective mental health system, the World Health Organization recommends the involvement of primary care in prevention and treatment of mild diseases and community-based care for serious mental illnesses. Despite a prevalence of lifetime mental health disorders above 30%, Portugal is failing to achieve such recommendations. It was argued that this failure is partly due to inadequate financing mechanisms of mental health care providers. This study proposes an innovative payment model for mental health providers oriented toward incentivising best practices. We performed a comprehensive review of healthcare providers' payment schemes and their related incentives, and a narrative review of best practices in mental health prevention and care. We designed an alternative payment model, on the basis of the literature, and then we presented it individually, through face-to-face interviews, to a panel of 22 experts with different backgrounds and experience, and from southern and northern Portuguese regions, asking them to comment on the model and provide suggestions. Then, after a first round of interviews, we revised our model, which we presented to experts again for their approval, and provide new suggestions and comments, if deemed necessary. This approach is close to what is generally known as the Delphi technique, although it was not applied in a rigid way. We designed a four-dimension model that focused on (i) the prevention of mental disorders early in life; (ii) the detection of mental disorders in childhood and adolescence; (iii) the implementation of a collaborative stepped care model for depression; and (iv) the integrated community-based care for patients with serious mental illnesses. First, we recommend a bundled payment to primary care practices for the follow-up of children with special needs or at risk under 2 years of age. Second, we propose a pay-for-performance scheme for all primary care practices, based on the number of users under 18 years old who are provided with check-up consultations. Third, we propose a pay-for-performance scheme for all primary care practices, based on the implementation of collaborative stepped care for depression. Finally, we propose a value-based risk-adjusted bundled payment for patients with serious mental illness. The implementation of evidence-based best practices in mental health needs to be supported by adequate payment mechanisms. Our study shows that mental health experts, including decision makers, agree with using economic tools to support best practices, which were also consensual.


Improving the wetting and dissolution of ibuprofen using solventless co-milling.
The wetting and dissolution of a BCS class II drug (Ibuprofen) is enhanced by solventless solid dispersion technique using co-milling. The co-milling is performed in a planetary ball mill using 1:1wt. ratio of Ibuprofen (drug) and Microcrystalline cellulose, MCC (excipient). The improvement in wettability and dissolution after co-milling are compared with the raw ibuprofen, ball-milled ibuprofen without any excipient and v-blend mixture of ibuprofen with an excipient. The changes in crystal level properties and reduction in crystallinity due to co-milling are measured using Powder X-ray diffraction (P-XRD) and Differential Scanning Calorimetry (DSC) respectively. The increased interaction between ibuprofen and MCC as well as hydrogen bond formation is confirmed by Fourier Transform Infrared Spectroscopy (FTIR). The morphological changes are observed by optical microscopy and Field Emission Scanning Electron Microscopy (FESEM). The miscibility of drug and excipient and evidence of formation of glassy solutions are demonstrated by Modulated Temperature Differential Scanning Calorimetry (MTDSC) and Raman microscopy. The surface energy and wetting properties are determined using Inverse Gas Chromatography (IGC) and sessile drop method respectively. The results show that co-milling generates defects, strain, and reduction in crystallite size (changes in crystal level properties) which are responsible for the improvement of wetting and dissolution (96% in 90min). Also, with increase in co-milling time, the polar surface energy increases and the hydrophobic ibuprofen drug surface transforms into hydrophilic surface due to increase in OH groups of MCC on the ibuprofen surface. The present work quantified all the above-mentioned parameters including the acidic and basic parameters of co-milled ibuprofen using IGC. The technique improves the wetting and dissolution of hydrophobic drugs. It can be very well extended to BCS class II and IV drugs in the presence of different hydrophilic excipients.


Gene of the month: BCOR.
BCL-6 transcriptional corepressor (BCOR) gene is located at Xp11.4 and encodes a protein which is involved in transcriptional repression in association with BCL-6 and epigenetic silencing through polycomb repressive complex 1 (PRC1). BCOR mutations are being identified in an increasing number of tumours which are diverse in their anatomical location and clinical setting. Interestingly, these tumours share similar and overlapping histological features, namely small round blue cell morphology and a myxoid background with delicate capillary channels. Clear cell sarcoma of the kidney, primitive myxoid mesenchymal tumour of infancy and central nervous system high-grade neuroepithelial tumour with BCOR alteration all share similar internal tandem duplications in the polycomb-group really interesting new gene (RING) finger homolog ubiquitin-likefold discriminator domain of BCOR Translocations resulting in BCOR fusion with CCNB3, MAML3 and ZC3H7B have been identified in undifferentiated round cell sarcoma. Subsets of high-grade endometrial stromal sarcoma and ossifying fibromyxoid tumour which have a more aggressive clinical course have been shown to harbour ZC3H7B-BCOR fusions. BCOR immunohistochemistry is an established marker with diagnostic utility.


Phase II biotransformation of carcinogens/atherogens in cultured aortic tissues and cells. I. Sulfation of 3-hydroxy-benzo(a)pyrene.
The capacity of polynuclear aromatic hydrocarbons to elicit profound effects on the development of avian aortic atheromata has raised questions regarding the biotransformation of polynuclear aromatic hydrocarbons in the target (aortic) tissue. Results of this investigation demonstrate the capacity of aortic enzymes to affect the sulfoconjugation of 3-hydroxy-benzo(a)pyrene and describe several characteristics of the aortic sulfotransferase activity. Conjugating activities measured in avian aortic tissues were approximately 10-20% of those assayed in corresponding preparations of avian hepatic tissues under the same reaction conditions. Activities were measured in homogenates, in a series of homogenate subfractions, in whole organ cultures, in cultured aortic endothelial cells, and in cultured aortic smooth muscle cell preparations. Sulfoconjugation was localized in the cytosolic fraction and kinetics in this fraction yielded a range of apparent Km values from 9 to 16 microM (mean = 11.8 +/- 3.1, n = 4) and a range of apparent Vmax values from 281 to 457 pmol/mg of protein/30 min (mean = 360 +/- 49, n = 4). Abdominal and thoracic segments of the aorta exhibited virtually identical specific activities. Also, activities assayed in cultured aortic smooth muscle cells were similar to those measured in cultured aortic endothelial cells. Capacity to generate adenosine 3'-phosphate 5'-phosphosulfate (PAPS) appeared to limit the reaction rate as judged by comparative investigations with PAPS and a PAPS-generating system. Aortic sulfatases actively hydrolyzed benzo(a)pyrene-3-O-sulfate. The sulfatase activity appeared to partially mask sulfotransferase activities measured in organ and cell culture preparations and in particulate subfractions of cellular homogenates.(ABSTRACT TRUNCATED AT 250 WORDS)