Protective Effect of Diphlorethohydroxycarmalol Isolated from Ishige okamurae Against Particulate Matter-Induced Skin Damage by Regulation of NF-κB, AP-1, and MAPKs Signaling Pathways In Vitro in Human Dermal Fibroblasts.
Particulate matters (PM), the main contributor to air pollution, have become a serious issue that threatens human's health. Skin is the largest organ in humans, as well as the primary organ exposed to PM. Overexposure of PM induces skin damage. Diphlorethohydroxycarmalol (DPHC), an algal polyphenol with the potential of skin protection, has been isolated from the edible brown seaweed Ishige okamurae. The purpose of the present study is to investigate the protective effect of DPHC against PM (ERM-CZ100)-induced skin damage in human dermal fibroblasts (HDF) cells. The results indicated that DPHC significantly and dose-dependently reduced intracellular reactive oxygen species generation in HDF cells. In addition, DPHC significantly induced collagen synthesis and inhibited collagenase activity in ERM-CZ100-stimulated HDF cells. Further study demonstrated that DPHC remarkably reduced the expression of human matrix metalloproteinases through regulation of nuclear factor kappa B, activator protein 1, and mitogen-activated protein kinases signaling pathways in ERM-CZ100-stimulated HDF cells. This study suggested that DPHC is a potential candidate to protect skins against PM-induced damage, and it could be used as an ingredient in pharmaceutical and cosmeceutical industries.


Isoaccepting lysine transfer ribonucleic acid species of Pseudomonas aeruginosa.
Pseudomonas aeruginosa tRNA was treated with iodine, CNBr and N-ethylmaleimide, three thionucleotide-specific reagents. Reaction with iodine resulted in extensive loss of acceptor activity by lysine tRNA, glutamic acid tRNA, glutamine tRNA, serine tRNA and tyrosine tRNA. CNBr treatment resulted in high loss of acceptor ability by lysine tRNA, glutamic acid tRNA and glutamine tRNA. Only the acceptor ability of tyrosine tRNA was inhibited up to 66% by N-ethylmaleimide treatment, a reagent specific for 4-thiouridine. By the combined use of benzoylated DEAE-cellulose and DEAE-Sephadex columns, lysine tRNA of Ps. aeruginosa was resolved into two isoaccepting species, a major, tRNA Lys1 and a minor, tRNALys1. Co-chromatography of 14C-labelled tRNALys1 and 3H-labelled tRNALys2 on benzoylated DEAE-cellulose at pH 4.5 gave two distinct, non-superimposable profiles for the two activity peaks, suggesting that they were separate species. The acceptor activity of these two species was inhibited by about 95% by iodine and CNBr. Both the species showed equal response to codons AAA and AAG and also for poly(A) and poly(A1,G1) suggesting that the anticodon of these species was UUU. Chemical modification of these two species by iodine did not inhibit the coding response. The two species of lysine of Ps. aeruginosa are truly redundant in that they are indistinguishable either by chemical modification or by their coding response.


[Calcifying tendinitis of the rotator cuff with focal umeral osteolysis. Imaging features.]
Calcifying tendinitis occurs most commonly in the rotator cuff tendons, particularly involving the supraspinatus tendon insertion, and is often asymptomatic. Cortical erosion secondary to calcifying tendinitis has been reported in multiple locations, including in the rotator cuff tendons. The authors report two cases of symptomatic calcifying tendinitis involving the infraspinatus tendon with cortical erosion with correlative radiographic, and MR findings. The importance of considering this diagnosis when evaluating lytic lesions of the humerus and the imaging differential diagnosis of calcifying tendinitis and cortical erosion are discussed.


Diverse and contrasting effects of habitat fragmentation.
Different components of an ecosystem can respond in very different ways to habitat fragmentation. An archipelago of patches, representing different levels of fragmentation, was arrayed within a successional field and studied over a period of 6 years. Ecosystem processes (soil mineralization and plant succession) did not vary with the degree of subdivision, nor did most measures of plant and animal community diversity. However, fragmentation affected vertebrate population dynamics and distributional patterns as well as the population persistence of clonal plant species. The results highlight the dangers of relying on broad community measures in lieu of detailed population analyses in studies of fragmented habitats.


Nicotine, acetylcholine and bombesin are trophic growth factors in neuroendocrine cell lines derived from experimental hamster lung tumors.
Neuroendocrine hamster lung tumors, induced by exposure to 60% hyperoxia and subcutaneous administration of the tobacco-specific nitrosamine 4-(methylnitrosamino)-l-(3-pyridyl)-l-butanone (NNK) for 12 weeks, were placed in cell culture. By subsequent selective transfer of epithelial cells and maintenance in an atmosphere of 8% CO2, cell lines with characteristics of neuroendocrine cells were established. The neuroendocrine markers expressed by these cell lines included electron dense neuroendocrine secretion granules as well as secretion of calcitonin and mammalian bombesin. In keeping with data previously reported for a human neuroendocrine lung tumor cell line, nicotine, acetylcholine, and mammalian bombesin (MB) acted as strong growth factors in these neuroendocrine hamster tumor lines. The mitogenic effect of nicotine and acetylcholine was abolished by nicotinic receptor inhibition while the effects of mammalian bombesin were inhibited by an antagonist of MB receptors. Our data suggest that a receptor-mediated mitogenic effect of nicotine on neuroendocrine lung cells may be instrumental in the induction of smoking-associated small cell lung cancer.


Segmental motions of rat thymidylate synthase leading to half-the-sites behavior.
Thymidylate synthase (TS) is a homodimeric enzyme with two equivalent active sites composed of residues from both subunits. Despite the structural symmetry of the enzyme, certain experimental results are consistent with half-the-sites activity, suggesting negative cooperativity between the active sites. To gain insight into the mechanism behind this phenomenon, we explore segmental motions of rat TS in the absence of ligands, with normal mode analysis as a tool. Using solvent accessible surface area of the active site pocket as a monitor of the degree of opening of the active sites, we classified the first 25 nontrivial normal modes, obtained from the web server of the program ElNémo, according to the behavior of the active sites. We found seven modes that open and close both sites symmetrically and nine that do so in an anticorrelated fashion. We characterized the motions of these modes by visual inspection and through measurement of distances between selected atoms lining the active site pockets. The segments that regulate access to the active site correspond to the loop containing R44, helix K, and a long loop containing residues 103-125, in agreement with a large body of crystallographic studies. These elements can be activated together or in isolation. There are more asymmetric modes than symmetric ones in the set we analyzed, probably accounting for the half-the-sites behavior of the enzyme. Three of the asymmetric modes result in changes at the dimer interface and indicate the endpoints of possible communication pathways between the active sites.


[Physicians' clinical skills after finishing internship].
The aim of this study was to evaluate Danish interns' clinical skills after their internship in order to identify areas where systematic training is needed. This study was a follow-up of a previous study on a group of interns at the beginning of their internship. A questionnaire listing 88 practical skills was posted to a group of trainees before and after internship. The interns were asked to rate their level of mastery in each skill on a scale 0-10, where 0 = not mastery/not competent and 10 = mastery/competent. In all, 74 out of 100 trainees returned the questionnaire. In 60 of the skills the trainees reported a higher level of mastery after internship and in 28 procedures the level of skill did not improve. Ten of these are procedures used in emergency settings. The reliability of self-reported clinical skills can be discussed, but it is a major concern that skills particularly in emergency procedures do not improve. There is a need to define clear objectives, implement structured training programmes, and assess skills in order to accomplish a reliable and sufficient level of competency.


Metabolism and pharmacological activities of the natural health-benefiting compound diosmin.
Diosmin is a famous natural flavonoid for treating chronic venous insufficiency and varicose veins. Recently, extensive study has indicated that diosmin possesses diverse pharmacological activities, including anti-inflammation, anti-oxidation, anti-diabetes, anti-cancer, anti-microorganism, liver protection, neuro-protection, cardiovascular protection, renoprotection, and retinal protection activities. Due to its low water solubility, diosmin is dramatically limited in clinical application. Expectedly, many potential strategies have been developed for improving its pharmacokinetic values and bioavailability. This health-benefiting compound has been explored as the major component of Daflon and micronized purified flavonoid fraction (MPFF), which have been used in clinics to improve micro-circulation. However, no specific drug targets for diosmin are reported, although some potential factors have been involved in screening, such as P-glycoprotein (P-gp), IKKβ, acetylcholinesterase (AChE), and aldose reductase (AR). More investigations on the underlying mechanisms of diosmin in mediating cellular processes with high specificity is still needed.


Cartilage tissue formation using redifferentiated passaged chondrocytes in vitro.
Articular cartilage has limited ability for repair when damaged by trauma or degenerative disease, such as osteoarthritis, which can result in pain and compromised quality of life. Biological surface replacements developed using tissue engineering methods are a promising approach for cartilage repair, which would avoid the need for total joint replacement with the synthetic implants used currently. A basic requirement of in vitro tissue generation is a supply of sufficient number of cells, which are difficult to acquire from sparsely cellular cartilage tissue. Previously, we have shown that coculture of in vitro-expanded dedifferentiated chondrocytes (P2) with small numbers of primary chondrocytes (P0) induces redifferentiation in passaged (P2) cells. In this study we show that this redifferentiation is not a transient change. After 4 weeks of coculture, the P0 and P2 cells were separated by flow-associated cell sorting, and the redifferentiated P2 (dP2) were cultured alone for a further 4 weeks. The redifferentiated dP2 cells formed thicker cartilage tissue compared to the tissue generated by P2 cells. The newly formed tissue contained type II collagen as demonstrated by immunohistochemical staining and accumulated more proteoglycan per cell than the tissue formed by P2 cells. The dP2 cells also exhibited higher type II collagen and lower type I collagen gene expression than the P2 cells. Interestingly, dP2 cells were able to exert the same effect as P0 cells when cocultured with P2 cells. In conclusion, under proper culture conditions, redifferentiated passaged chondrocytes behave similarly to primary chondrocytes. This coculture system approach can be used to increase the number of differentiated chondrocytes that can be obtained by classical monolayer cell expansion and represents a novel way to acquire sufficient cell numbers for cartilage tissue engineering.


Going for brokerage: strategies and strains in commercial healthcare facilitation.
The formation of domestic and global marketplaces during the past 50 years has opened up new commercial opportunities for third-party activity in healthcare systems. Commercial mediation of access to healthcare is one recent area of activity that sees companies and individuals offering to organise healthcare and travel in return for payment. With varying degrees of control over the location, type, cost and experiences of healthcare provisioning, these intermediaries occupy potentially influential positions in healthcare systems and yet much of their work is poorly understood. Drawing on social science theories of brokerage, this article presents a novel analysis of commercial healthcare facilitation. It focuses on facilitation companies and their workers as central, intermediating actors for people to access healthcare in markets characterised by complexity. Semi-structured interviews were conducted with people working in domestic and international healthcare facilitation in London and Delhi, and data were analysed using a framework approach that emphasises the structural features and personal agencies for this area of work. Findings point to an institutional environment for commercial healthcare facilitation marked by competition and the threat of obsolescence. The activities of rivals, and the risk that users and providers will bypass intermediaries, compels facilitation companies to respond strategically and to continuously pursue new populations and activities to mediate - to go for broke. These pressures percolate into the lives of people who perform facilitation work and who describe a physical and mental burden of labour incurred by onerous processes for generating and completing facilitation work. The need for language interpretation services introduces an additional set of relations and has created further points of tension. It is an environment that engenders mistrust and anxiety, and which incentivises exploitation and a commodification of users whose associated commissions are highly prized. Brokerage analysis provides valuable insights into the strategies and strains for commercial mediation of access to healthcare, and the findings indicate opportunities for further research on the contributions of interpreters, diplomatic and business networks, and new technologies, and on the growth of new forms of mediation in domestic and overseas settings.


Myocardial changes in influenza virus and staphylococcus infection.
Experiments carried out on 134 newborn mice showed that influenza virus and staphylococcus infection and their combination induces local necrosis in the myocardium. Morphometric measurement has shown that the extent of necrotic changes amounted with the action of the influenza virus to 1.37%, with the action of staphylococcus to 1.4% and with their combined action to 1.8%. Electron microscopy revealed dystrophic changes in cardiomyocytes with subsequent intracellular regeneration.


Coronary Vasospasm After Burn Injury: First Described Case Series of a Lethal Lesion.
Burn injuries generate multisystem physiological derangements. The authors present a case series of three patients developing acute coronary syndrome (ACS) stemming from coronary vasospasm (CVS) over a course of 5 months. This etiology of ACS is significant as it has previously not been described in burn patients and requires a different management algorithm than vaso-occlusive disease.All patients were admitted to a single accredited burn unit. Burn mechanisms were flash burn, chemical fire, and house fire. TBSA were 20%, 72%, and 31%, respectively. Ages were 67-, 41-, and 52-year-old men. All smoked tobacco, and one had diabetes and coronary artery disease. CVS presented with acute onset ST elevations, severe bradycardia, and cardiac arrest in all. Diagnosis was made via EKG and angiography. Treatment was undertaken with nitrates and calcium channel blockers. One of the patients died of refractory disease.The authors identified CVS in burn patients with an incidence of 2% of admissions and accounting for 17% of our burn mortality during this period. To date, there is no linkage between CVS and burns described in humans; however, there is a well-described animal model in rats. The risk factors for CVS are common among burn patients, particularly smoking and endothelial dysfunction. CVS may be a significant cause of ACS in burn patients, and it should be considered in the setting of ACS especially with a negative angiography. Knowledge of this disease state can change burn management to mitigate risk and accommodate specific cardiac treatments.


Intermittent restraint-induced sympathetic activation attenuates hepatic steatosis and inflammation in a high-fat diet-fed mouse model.
Nonalcoholic fatty liver disease (NAFLD) is very prevalent worldwide and is associated with insulin resistance and metabolic syndrome. Stress is a physiological and biological response to maintain homeostasis of the body against stressors while severe stress response is an important contributor to various illnesses, including metabolic syndrome and brain disorders. We have evaluated the effects of intermittent restraint stress on NAFLD in a high-fat diet (HFD)-fed mouse model. C57/BL6 mice had free access to a 60% HFD for 8 wk, with or without intermittent restraint stress (3 h) conducted three times a week. HFD administration increased fat accumulation in liver tissues. Unlike the stressed standard diet group, the levels of hepatic total cholesterol and triglycerides were significantly ameliorated in the HFD with stress group compared with the HFD alone group. These beneficial results were in accordance with serum levels of liver enzymes (aspartate transaminase, alanine transaminase) and hepatic levels of TNF-α and oxidative stress parameters (reactive oxygen species, nitric oxide, and malondialdehyde). The intermittent restraint stress significantly attenuated the HFD-derived alterations in serum insulin levels, hepatic protein kinase B activity, and gene expression, especially related to lipogenesis. This intermittent restraint stress also elevated the serum epinephrine concentration and activated the adrenergic receptor β2 or β3 in livers or white adipose tissue (WAT). Activation of energy expenditure markers (uncoupling protein 1, peroxisome proliferator-activated receptor-γ coactivator-1α) in brown adipose tissue and the browning of WAT were also observed in the HFD with stress group. Taken together, our findings showed the beneficial effects of sympathetic activation by intermittent restraint stress on HFD-induced hepatic steatosis and partial inflammation.NEW & NOTEWORTHY In modern society, stress is a part of daily life, and a certain level of stress is inevitable to most of the general population. Uncontrolled severe stress is obviously harmful; however, certain kind/level of stress could be beneficial on lipid metabolism via sympathetic activation. Our data suggest that a sympathetic activation by intermittent restraint stress could play a positive role in maintaining the balance of hepatic lipid metabolism, especially under high-fat diet conditions.


Urachal Remnant Infection Caused by Actinomyces Turicensis.
Urachal remnants (URs) represent uncommon and underdiagnosed entities that are usually detected incidentally at imaging or present clinically different manifestations. Here we presented a boy with UR infection. Ultrasonography and bacterial culture and identification were performed. He received antibiotic treatment and underwent surgical excision of the cyst. The patient presented with both a urachal cyst and umbilical-urachal sinus. UR infection was caused by Actinomyces turicensis. He recovered well from the operation without complications. The present case reminds clinicians to be familiar with imaging features of different types of URs and their potential complications and indicates the necessity of pathogenic microorganism analysis to tailor antibiotic treatment and post-operative follow-up to prevent complications.


Serum surfactant protein D as a marker for bronchopulmonary dysplasia.
Lung epithelial cells express surfactant protein D (SP-D), a calcium-dependent lectin that plays an important role in antibody-independent pulmonary host defense. Previous studies have shown that it is found in the peripheral circulation in patients with pulmonary disease, likely because of translocation into the blood when lung epithelial barriers are disrupted by inflammation or acute injury. In adults, serum SP-D levels are biomarkers for the progression and severity of chronic lung disease. In neonates, elevated SP-D levels in cord blood and on day 1 have been associated with prenatal risk factors and with an increased risk of respiratory distress syndrome and infections. It is not known whether serum SP-D during the first week of life is a marker for bronchopulmonary dysplasia (BPD), a form of chronic lung disease of prematurity that is associated with lung parenchymal maldevelopment and injury. The goal of this study is to determine whether serum SP-D on days 3 and 7 of life are associated with the development of BPD in preterm infants. Serum samples were obtained on postnatal days 3 and 7 from 106 preterm infants (500-2000 g birth weight, 23-32-week gestation). SP-D was quantified by Western blot. BPD was determined at 36 weeks PMA using NICHD criteria. The mean birth weight was 1145 ± 347 g and gestational age 29.2 ± 7.4 weeks. BPD was diagnosed in 7 and "BPD or death" in 16 infants. Days 3 and 7 values tracked significantly (r = 0.648), and did not correlate with birth weight or gestational age. Contrary to expectations, serum SP-D was not associated with BPD. Significant gender differences were noted, with SP-D dropping from day 3 to day 7 in males, while increasing in females (p < .05). Elevated serum SP-D does not appear to be a useful marker for BPD. Decreasing serum SP-D levels in males, as compared to females, during the first week of life are likely related to gender differences in lung maturation, consistent with the higher incidence of BPD in males.


Transmission dosimetry with a liquid-filled electronic portal imaging device.
To assess the accuracy of transmission dose rate measurements for various phantom-detector geometries, performed with an electronic portal imaging device (EPID) and to compare these transmission dose rate values with exit dose rate data. Transmission dose rate values on the central beam axis and beam profiles were measured with an EPID consisting of a matrix of liquid-filled ionization chambers. These data were compared with transmission and exit dose rate values, obtained using air-filled ionization chambers for a number of field sizes, phantom thickness, and phantom-detector distances. Various homogeneous and inhomogeneous phantoms were applied. The increase in dose rate with field size is larger for the EPID than in air, due to the larger amount of side scatter in the EPID. The difference has been taken into account by a deconvolution of the EPID images. An additional build-up layer on top of the commercial device is needed to reach dose maximum at the liquid ionization chambers for photon beam energies higher than about 4 MV. The transmission off-axis ratios (OAR) determined with the EPID and in air agreed within 2% for all tested cases, after deconvolution of the EPID signal. The agreement between the EPID-and exit-OAR decreased with increasing phantom-detector distance and the presence of inhomogeneities. For a phantom-detector distance of about 10 cm, the EPID- and exit-OARs agree within 2.5%. The difference could be up to 8% for an air inhomogeneity and a phantom-detector distance of 30 cm. The difference between EPID measurements and measurements in air can be explained by side scatter effects in the EPID and lack of adequate buildup, and can easily be taken into account. The loss of scatter compared with the situation at the exit side of the phantom explains the difference between transmission and exit dose values. At short phantom-detector distances, good agreement exists between transmission and exit dose rate. This implies that at this distance, the EPID can be used for simple comparison with exit dose calculations during patient treatments. At larger distances, more sophisticated conversion methods are required.


Level of satisfaction in patients with maxillary full-arch fixed prostheses: zygomatic versus conventional implants.
To evaluate the satisfaction of patients with maxillary fixed prostheses supported by conventional and/or zygomatic implants. Patients restored with complete maxillary implant-supported fixed prostheses were included in the study. They were divided into 2 groups: those with at least 1 zygomatic implant (the zygomatic group) and those with no zygomatic implants (the nonzygomatic group). Twelve months after prosthesis delivery, subjects indicated their satisfaction with the new prosthesis on a visual analog scale. Prostheses were rated in terms of general satisfaction, comfort and stability, ability to speak, easy of cleaning, esthetics, self-esteem, and functionality. Forty-six patients participated in the study (23 in each group). The mean level of satisfaction was high; the groups differed significantly only in satisfaction with esthetics. Patients in the zygomatic group had a higher average score for esthetics than those in the nonzygomatic group. Those who had previously worn conventional removable prostheses gave a higher score for functionality to the implant-supported fixed prosthesis compared to the removable prosthesis. Patient satisfaction with zygomatic implant-supported fixed prostheses was similar to that for fixed prostheses supported by conventional implants.


Barriers, refusal and a hypothetical monetary incentive for Covid-19 vaccination in Mexican adults.
To estimate vaccine uptake and assess sociodemographic conditions associated with vaccination barriers and refusal and to explore the effect of a monetary incentive to overcome them. We used data from adults from the 2021 National Continuous Health and Nutrition Survey conducted during August-October 2021. We evaluated if an hypothetical monetary incentive between 50-650 MXN (~2.5-31 USD) would overcome barriers or refusal. 73.9% were vaccinated with at least one dose, 7.5% refused, 4.8% reported barriers and 13.8% were ineligible at the time of the survey. Refusal and barriers were more frequent in men, older age, lower education and socioeconomic status, unemployed and informal workers. In people with barriers and refusal, the hypothetical incentive increased the acceptance in 57.6% (95%CI 50.7,64.4%) and 17.4% (95%CI 13.2,21.7%) in people with barriers and refusal, respectively. Understanding the reasons for barriers and refusal is crucial for future Covid-19 vaccination campaigns or epidemics. A monetary incentive might increase vaccination uptake, although, cost-effectiveness analyses are needed.


Management of Zygomatic Fixture Complication Case Using Extra-Short Implants.
The oral rehabilitation of a patient with atrophic maxilla is one of the main challenges in implant-supported rehabilitations. The implant placement in the posterior region of the maxilla is limited by a deficiency of bone quantity and quality, besides the anatomic restriction of the maxillary sinus. The use of zygoma fixtures, anchoraged in the zygomatic bone, is a useful technique to enable the full arch rehabilitation. However, complications related to this procedure bring a lot of difficulty in solving. This study presents the use of extra-short implants as a strategy to treat a patient in whom the zygoma fixture and the fixed prosthesis failed, associated with sinusitis and alveolar bone loss.


Newborn splenic volumes vary under different malaria endemic conditions.
Ultrasound was used to measure newborn splenic dimensions and calculate the volumes in a malarious and a non-malarious region of Papua New Guinea. The median splenic volume of infants born in Madang, where malaria transmission is high throughout the year, was 5.2 cm3/kg, while that of infants born in Goroka, where malaria is not endemic, was 2.6 cm3/kg. The cause of this difference is unknown, but possible explanations include fetal exposure to malaria antigens in utero and the high incidence of inherited red cell disorders in the malarious regions of Papua New Guinea.


Analysis and evolution of health policies in Iran through policy triangle framework during the last thirty years: a systematic review of the historical period from 1994 to 2021.
Health policy analysis as a multi-disciplinary approach to public policy illustrates the need for interventions that highlight and address important policy issues, improve the policy formulation and implementation process and lead to better health outcomes. Various theories and frameworks have been contributed as the foundation for the analysis of policy in various studies. This study aimed to analyze health policies during the historical period of the almost last 30 years in Iran using policy triangle framework. To conduct the systematic review international databases (PubMed / Medline, Scopus, Web of Sciences, CINAHL, PsycINFO, Embase, the Cochran Library) and Iranian databases from January 1994 to January 2021 using relevant keywords. A thematic qualitative analysis approach was used for the synthesis and analysis of data. The Critical Appraisal Skills Programme for Qualitative Studies Checklist (CASP) was conducted. Out of 731 articles, 25 articles were selected and analyzed. Studies used health policy triangle framework to analyze policies in the Iranian health sector has been published since 2014. All the included studies were retrospective. The main focus of most of studies for the analysis was on the context and process of polices as the elements of the policy triangle. The main focus of health policy analysis studies in Iran over the last thirty years was on the context and process of polices. Although range of actors within and outside the Iran government influence health policies but in many policy processes the power and the role of all actors or players involved in the policy are not recognized carefully. Also, Iran's health sector suffers from lack of a proper framework for evaluating various implemented policies.


Protein folding causes an arrest of preprotein translocation into mitochondria in vivo.
With vital yeast cells, a hybrid protein consisting of the amino-terminal third of the precursor to cytochrome b2 and of the entire dihydrofolate reductase was arrested on the import pathway into mitochondria. Accumulation of the protein in the mitochondrial membranes was achieved by inducing a stable tertiary structure of the dihydrofolate reductase domain. Thereby, three salient features of mitochondrial protein uptake in vivo were demonstrated: its posttranslational character; the requirement for unfolding of precursors; and import through translocation contact sites. The permanent occupation of translocation sites by the fusion protein inhibited the import of other precursors; it did, however, not lead to leakage of mitochondrial ions, implying the existence of a channel that is sealed around the membrane spanning polypeptide segment.


Europium-Diethylenetriaminepentaacetic Acid Loaded Radioluminescence Liposome Nanoplatform for Effective Radioisotope-Mediated Photodynamic Therapy.
Photodynamic therapy (PDT) is an effective anticancer strategy with a higher selectivity and fewer adverse effects than conventional therapies; however, shallow tissue penetration depth of light has hampered the clinical utility of PDT. Recently, reports have indicated that Cerenkov luminescence-induced PDT may overcome the tissue penetration limitation of conventional PDT. However, the effectiveness of this method is controversial because of its low luminescence intensity. Herein, we developed a radiolabeled diethylenetriaminepentaacetic acid chelated Eu3+ (Eu-DTPA)/photosensitizer (PS) loaded liposome (Eu/PS-lipo) that utilizes ionizing radiation from radioisotopes for effective in vivo imaging and radioluminescence-induced PDT. We utilized Victoria blue-BO (VBBO) as a PS and observed an efficient luminescence resonance energy transfer between Eu-DTPA and VBBO. Furthermore, 64Cu-labeled Eu lipo demonstrated a strong radioluminescence with a 2-fold higher intensity than Cerenkov luminescence from free 64Cu. In our radioluminescence liposome, radioluminescence energy transfer showed a 6-fold higher energy transfer efficiency to VBBO than Cerenkov luminescence energy transfer (CLET). 64Cu-labeled Eu/VBBO lipo (64Cu-Eu/VBBO lipo) showed a substantial tumor uptake of up to 19.3%ID/g by enhanced permeability and retention effects, as revealed by in vivo positron emission tomography. Finally, the PDT using 64Cu-Eu/VBBO lipo demonstrated significantly higher in vitro and in vivo therapeutic effects than Cerenkov luminescence-induced PDT using 64Cu-VBBO lipo. This study envisions a great opportunity for clinical PDT application by establishing the radioluminescence liposome which has high tumor targeting and efficient energy transfer capability from radioisotopes.


Noisy continuous time random walks.
Experimental studies of the diffusion of biomolecules within biological cells are routinely confronted with multiple sources of stochasticity, whose identification renders the detailed data analysis of single molecule trajectories quite intricate. Here, we consider subdiffusive continuous time random walks that represent a seminal model for the anomalous diffusion of tracer particles in complex environments. This motion is characterized by multiple trapping events with infinite mean sojourn time. In real physical situations, however, instead of the full immobilization predicted by the continuous time random walk model, the motion of the tracer particle shows additional jiggling, for instance, due to thermal agitation of the environment. We here present and analyze in detail an extension of the continuous time random walk model. Superimposing the multiple trapping behavior with additive Gaussian noise of variable strength, we demonstrate that the resulting process exhibits a rich variety of apparent dynamic regimes. In particular, such noisy continuous time random walks may appear ergodic, while the bare continuous time random walk exhibits weak ergodicity breaking. Detailed knowledge of this behavior will be useful for the truthful physical analysis of experimentally observed subdiffusion.


Love hurts: an fMRI study.
Being in a close relationship is essential to human existence. Such closeness can be described as including other in the self and be underpinned on social attachment system, which evolved from a redirection of nociceptive mechanisms. To what extent does imagining a loved-one differs from imagining an unfamiliar individual being in painful situations? In this functional MRI study, participants were exposed to animated stimuli depicting hands or feet in painful and non-painful situations, and instructed to imagine these scenarios from three different perspectives: self, loved-one and stranger after being primed with their respective photographs. In line with previous studies, the three perspectives were associated with activation of the neural network involved in pain processing. Specifically, adopting the perspective of a loved-one increased activity in the anterior cingulate cortex and insula, whereas imagining a stranger induced a signal increase in the right temporo-parietal junction (TPJ) and superior frontal gyrus. The closer the participants' relationships were with their partner, the greater the deactivation in the right TPJ. A negative effective connectivity between the right TPJ and the insula, and a positive one with the superior frontal gyrus were found when participants imagined the perspective of a stranger. These results demonstrate that intimacy affects the bottom-up information processing involved in empathy, as indicated by greater overlap between neural representations of the self and the other.


Dietary sodium loading increases plasma brain natriuretic peptide levels in man.
The effect of dietary sodium loading on plasma human brain natriuretic peptide-like immunoreactivity (hBNP-li) was examined in 11 normotensive subjects aged 20-23 years. Plasma hBNP-li increased significantly with increasing dietary sodium intake, with levels of 1.33 +/- 0.17 pmol/l on day 5 of a normal-sodium diet (24-h urinary sodium excretion of 171 +/- 16 mmol) and 2.04 +/- 0.10 pmol/l (P less than 0.05, versus normal-sodium diet) on day 5 of a high-sodium diet (24-h urinary sodium excretion 503 +/- 36 mmol). Corresponding plasma atrial natriuretic factor levels were 5.6 +/- 1.7 pmol/l and 11.0 +/- 2.0 pmol/l (P less than 0.05, versus normal-sodium diet) on the normal- and high-sodium diets, respectively. These results suggest that, in addition to atrial natriuretic factor, BNP may be a new and important natriuretic peptide which regulates sodium homeostasis in man during increased sodium intake.


Vital warmth and well-being: steambathing as household therapy among the Tzeltal and Tzotzil Maya of highland Chiapas, Mexico.
Among the Maya, the cultural history of steambathing spans more than two millennia. Although it has largely disappeared from the lowlands, household-level steambathing persists in several highland Maya communities in Chiapas, Mexico. In this article, I present an overview of therapeutic steambathing among the Tzeltal and Tzotzil Maya. Through an extended discussion of the beliefs and practices surrounding steambathing, I develop several features of highland Maya thinking about physical health and "well-being". In particular, I examine a set of ethnophysiological representations relating to the "thermal" nature of functional bodies, and the relationship of these models to the maintenance and restoration of health. The highland Maya have articulated an elaborate understanding of physical health and well-being coded in an idiom of "vital warmth", and directed toward the preservation and augmentation of the endogenous heat necessary for vitality and vigor. These models simultaneously reflect empirical understandings of bodily states in health and illness, as well as metaphorical assumptions about the thermal nature of functional psychosocial identities. Steambathing draws on and reinforces these models, constituting a core cultural technology for radically altering the thermal state of the patient, an experience which the highland Maya regard as deeply beneficial. The paper closes with a discussion of recent biomedical research into the physiological effects of hyperthermal therapies.


Adverse social determinants and risk for congenital anomalies.
Different studies have related familiar and regional adverse social conditions to perinatal outcome (neonatal mortality, low birth weight and prematurity); however, few studies have studied the effect of poverty on congenital anomalies. To assess the hazard ratio of 25 congenital anomalies and adverse social determinants as per the socioeconomic level of families and regions. Exploratory, case-control study using data from the Latin-American Collaborative Study of Congenital Malformations (Estudio Colaborativo Latinoamericano de Malformaciones Congenitas, ECLAMC). The sample consisted of 3786 live newborn infants with a single malformation and 13,344 controls selected among 546,129 births occurred in 39 hospitals from Argentina in the 1992-2001 period. Both direct and indirect (residence) risks (OR) were estimated, together with the interaction between the individual and residential socioeconomic levels for each of the 25 congenital anomalies. Cleft lip with/without cleft palate (OR= 1.43) and ventricular septal defect (OR= 1.38) showed a significantly higher risk in the lower socioeconomic level. Low socioeconomic levels were significantly associated with a higher frequency of parental sibship (blood relationship); native descent; maternal age younger than 19 years old; more than four pregnancies; a low number of antenatal care visits; and residence in deprived regions. Cleft lip with/without cleft palate and ventricular septal defects were significantly associated with a lower socioeconomic level. Lack of family planning and antenatal care; and exposure to environmental or teratogenic agents may account for these findings.


EDL/MAE regulates EGF-mediated induction by antagonizing Ets transcription factor Pointed.
Inductive patterning mechanisms often use negative regulators to coordinate the effects and efficiency of induction. During Spitz EGF-mediated neuronal induction in the Drosophila compound eye and chordotonal organs, Spitz causes activation of Ras signaling in the induced cells, resulting in the activation of Ets transcription factor Pointed P2. We describe developmental roles of a novel negative regulator of Ras signaling, EDL/MAE, a protein with an Ets-specific Pointed domain but not an ETS DNA-binding domain. The loss of EDL/MAE function results in reduced number of photoreceptor neurons and chordotonal organs, suggesting a positive role in the induction by Spitz EGF. However, EDL/MAE functions as an antagonist of Pointed P2, by binding to its Pointed domain and abolishing its transcriptional activation function. Furthermore, edl/mae appears to be specifically expressed in cells with inducing ability. This suggests that inducing cells, which can respond to Spitz they themselves produce, must somehow prevent activation of Pointed P2. Indeed hyperactivation of Pointed P2 in inducing cells interferes with their inducing ability, resulting in the reduction in inducing ability. We propose that EDL/MAE blocks autocrine activation of Pointed P2 so that inducing cells remain induction-competent. Inhibition of inducing ability by Pointed probably represents a novel negative feedback system that can prevent uncontrolled spread of induction of similar cell fates.


Miniature photonic-crystal hydrophone optimized for ocean acoustics.
This work reports on an optical hydrophone that is insensitive to hydrostatic pressure, yet capable of measuring acoustic pressures as low as the background noise in the ocean in a frequency range of 1 Hz to 100 kHz. The miniature hydrophone consists of a Fabry-Perot interferometer made of a photonic-crystal reflector interrogated with a single-mode fiber and is compatible with existing fiber-optic technologies. Three sensors with different acoustic power ranges placed within a sub-wavelength sized hydrophone head allow a high dynamic range in the excess of 160 dB with a low harmonic distortion of better than -30 dB. A method for suppressing cross-coupling between sensors in the same hydrophone head is also proposed. A prototype was fabricated, assembled, and tested. The sensitivity was measured from 100 Hz to 100 kHz, demonstrating a sound-pressure-equivalent noise spectral density down to 12 μPa/Hz(1/2), a flatband wider than 10 kHz, and very low distortion.


Tobacco, Nicotine and Health.
Recent studies have explored improvements in smoking prevention and cessation to reduce smoking prevalence, however, in Europe, only Finland has already set a date to become nicotine free. Studies reporting on central, eastern and southern Europe have mostly focused on combustible cigarettes up to now. In young people, correlations were found between traditional smoking, the "vaping" of e-cigarettes, experimentation with alcohol intoxication, and the use of illicit drugs. Prevention and cessation should include strategies against active and passive exposures to new nicotine products. This is a prerequisite for a successful public health policy and a future end-game against the business interests of the tobacco industry and its allies.


[Morphological manifestations of the immune reactions in stomach cancer and their relation to the histological structure of the tumor].
The morphology of immune response in tumor tissue was studied in the resected material obtained from 244 patients with stomach cancer. In nearly 80% of cases of carcinoma, the following five morphological types were observed: (1) acute inflammation (immediate--onset hyperergy); (2) time--delayed hyperergy; (3) plasmocytic--eosinophilic cellular infiltration with tumor mucination; (4) chronic granulomatous inflammation and (5) lymphadenoid and fibrose tissue envelope of tumor. There established a relationship between the type and degree of immune reactions and the histology of tumor. Progressing, regressing and stable reactions should be distinguished. Tumor regression and immune response were observed in foci; however, complete resolution was registered in 2.5% of cases only.


[Inactivation of choline receptors induced by membrane depolarization].
It was found that membrane depolarization of Lymnaea stagnalis neurons inhibited the conductance induced by acetylcholine (ACh). This phenomenon (cholinoreceptor inactivation) was shown to be connected with a potential influence on none of the following characteristics: reversal potential (Er) of the ACh-induced currents, affinity of ACh to the receptors, ACh concentration near the receptive membrane surface, and desensitization rate.


src-related protein tyrosine kinases and their surface receptors.
The CD4-p56lck and CD8-p56lck complexes have served as a paradym for an expanding number of interactions between src-family members (p56lck, p59fyn, p56lyn, p55blk) and surface receptors. These interactions implicate src-related kinases in the regulation of a variety of intracellular events, from lymphokine production and cytotoxicity to the expression of specific nuclear binding proteins. Different molecular mechanisms appear to have evolved to facilitate the receptor-kinase interactions, including the use of N-terminal regions, SH2 regions and kinase domains. Variation exists in stoichiometry, affinity and the nature of signals generated by these complexes in cells. The CD4-p56lck complex differs from receptor-tyrosine kinases in a number of important ways, including mechanisms of kinase domain regulation and recruitment of substrates such as PI 3-kinase. Furthermore, they may have a special affinity for receptor-substrates such as the TcR zeta, MB1/B29 or CD5 receptors, and act to recruit other SH2-carrying proteins, such as ZAP-70 to the receptor complexes. Receptor-src kinase interactions represent the first step in a cascade of intracellular events within the protein-tyrosine kinase/phosphatase cascade.


Impact and shear resistance of turf grass racing surfaces for Thoroughbreds.
The 2 dominant processes involved in the horse hoof-to-racing surface interaction are the shock loading of the horse's leg upon impact with the racing surface and rotation of the horse's hoof into the racing surface. These processes were measured as impact resistance (ie, the peak deceleration of a moving body upon impact with the test surface) and as resistance to shear. The objective of the present study was to measure physically (under a variety of conditions) those soil and turf factors related to the 2 processes. It was concluded that thatch accumulation and mowing height of the turf did not have a significant (P less than 0.05) effect on racing surface hardness (therefore, turf management should optimize turf growth and recovery), that turf roots were responsible for an increase in impact resistance and in resistance to shear, that control over soil moisture through irrigation and drainage allowed modification of racing surface hardness, and that soil materials tend to have lower impact resistance (ie, lower shock loading of the horse's leg) and higher resistance to shear (ie, greater resistance to hoof rotation) than do sand materials.


Complement receptor-3 negatively regulates the phagocytosis of degenerated myelin through tyrosine kinase Syk and cofilin.
Intact myelin, which normally surrounds axons, breaks down in Wallerian degeneration following axonal injury and during neurodegenerative diseases such as multiple sclerosis. Clearance of degenerated myelin by phagocytosis is essential since myelin impedes repair and exacerbates damage. CR3 (complement receptor-3) is a principal phagocytic receptor in myelin phagocytosis. We studied how tyrosine kinase Syk (spleen tyrosine kinase) and cofilin control phagocytosis of degenerated myelin by CR3 in microglia and macrophages. Syk is a non-receptor tyrosine kinase that CR3 recruits to convey cellular functions. Cofilin is an actin-depolymerizing protein that controls F-actin (filamentous actin) remodeling (i.e., disassembly and reassembly) by shifting between active unphosphorylated and inactive phosphorylated states. Syk was continuously activated during prolonged phagocytosis. Phagocytosis increased when Syk activity and expression were reduced, suggesting that normally Syk down regulates CR3-mediated myelin phagocytosis. Levels of inactive p-cofilin (phosphorylated cofilin) decreased transiently during prolonged phagocytosis. In contrast, p-cofilin levels decreased continuously when Syk activity and expression were continuously reduced, suggesting that normally Syk advances the inactive state of cofilin. Observations also revealed inverse relationships between levels of phagocytosis and levels of inactive p-cofilin, suggesting that active unphosphorylated cofilin advances phagocytosis. Active cofilin could advance phagocytosis by promoting F-actin remodeling, which supports the production of membrane protrusions (e.g., filopodia), which, as we also revealed, are instrumental in myelin phagocytosis. CR3 both activates and downregulates myelin phagocytosis at the same time. Activation was previously documented. We presently demonstrate that downregulation is mediated through Syk, which advances the inactive phosphorylated state of cofilin. Self-negative control of phagocytosis by the phagocytic receptor can be useful in protecting phagocytes from excessive phagocytosis (i.e., "overeating") during extended exposure to particles that are destined for ingestion.


Low-level laser therapy ameliorates disease progression in a mouse model of multiple sclerosis.
Multiple sclerosis (MS) is an autoimmune demyelinating inflammatory disease characterized by recurrent episodes of T cell-mediated immune attack on central nervous system (CNS) myelin, leading to axon damage and progressive disability. The existing therapies for MS are only partially effective and are associated with undesirable side effects. Low-level laser therapy (LLLT) has been clinically used to treat inflammation, and to induce tissue healing and repair processes. However, there are no reports about the effects and mechanisms of LLLT in experimental autoimmune encephalomyelitis (EAE), an established model of MS. Here, we report the effects and underlying mechanisms of action of LLLT (AlGaInP, 660 nm and GaAs, 904 nm) irradiated on the spinal cord during EAE development. EAE was induced in female C57BL/6 mice by immunization with MOG35-55 peptide emulsified in complete Freund's adjuvant. Our results showed that LLLT consistently reduced the clinical score of EAE and delayed the disease onset, and also prevented weight loss induced by immunization. Furthermore, these beneficial effects of LLLT seem to be associated with the down-regulation of NO levels in the CNS, although the treatment with LLLT failed to inhibit lipid peroxidation and restore antioxidant defense during EAE. Finally, histological analysis showed that LLLT blocked neuroinflammation through a reduction of inflammatory cells in the CNS, especially lymphocytes, as well as preventing demyelination in the spinal cord after EAE induction. Together, our results suggest the use of LLLT as a therapeutic application during autoimmune neuroinflammatory responses, such as MS.


Quantitation of pilocarpine delivery across isolated rabbit cornea by noncross-linked high viscosity polymer gel.
The effect on pilocarpine flux across rabbit cornea in vitro by a noncross-linked polymeric gel vehicle was measured. A closed system transport chamber was used. Its design featured continuous flow of a tear analog but excluded variables of the internal eye. Results were compared to previously determined data in the same chamber system for cross-linked hydrogel buttons and for free pilocarpine fluid. Gel-mediated flux was equal to that with lens buttons to 90 minutes in the case of a 30 per cent gel (viscosity approximately 70,000 centipoises). Elution by the tear analog system limited flux duration of gels relative to lenses. Greater viscosity of 30 per cent gel relative to 25 per cent gel (approximately 15,000 centipoises) was associated with prolonged transcorneal drug flux. The congruence of flux slopes for 30 per cent gel and lens button vehicles despite the difference in available dose suggests saturable mediation of pilocarpine transport across the cornea, but a greater "flux efficiency" through 90 minutes for 30 per cent gel.


[Evaluation of a new blood pressure cuff for patients with large and normal arm circumferences].
A new multisize blood pressure cuff (TriCuff; Inmed) which automatically adjusts to fit any size arm was tested against the routinely used 12 x 23 cm cuff in patients with an arm circumference less than 33 cm (group A = 30) and against the 15 x 33 cm cuff as well as the 12 x 23 cm cuff in patients with arm circumference greater than 33 cm (group B = 26). Arm circumference in group A was 25 +/- 3 cm and in group B 38 +/- 3 cm (P less than 0.01). Systolic blood pressure (SBP) and diastolic blood pressure (DBP) in group A was 125 +/- 20 mmHg and 78 +/- 11 mmHg respectively with the TriCuff and 124 +/- 19 mmHg and 78 +/- 13 mmHg with the 12 x 23 cm cuff. In group B SBP and DBP with the TriCuff was 134.6 +/- 13.3 mmHg and 84 +/- 17 mmHg respectively compared with 132 +/- 15 mmHg and 83.8 +/- 16 mmHg measured by the 15 x 33 cm cuff. SBP and DBP with the 12 x 23 cm cuff was 139.8 +/- 16 mmHg and 90.8 +/- 14 mmHg respectively. Both these values are significantly higher (P less than 0.05) compared with the new cuff and the larger cuff. Using the 12 x 23 cm cuff in group B resulted in the false diagnosis of hypertension in 3 normotensive patients. The TriCuff gives accurate blood pressure measurements in patients with different arm circumferences and should prevent false-positive diagnoses of hypertension in obese patients.


[The diagnosis and treatment of odontogenic mediastinitis].
A program of early diagnostics of odontogenic mediastinitis was developed on the basis of an analysis of clinico-laboratory manifestations and the evaluation of its clinical efficiency is given. The criteria were established for determining the phases of odontogenic mediastinitis-reactive, toxic and terminal ones. A program of complex treatment was worked out and introduced into practice which included the methods of efferent therapy-hemosorption and perfusion through xenospleen. Substantially decreased lethality (from 39.5% to 15.3%) from odontogenic mediastinitis was obtained.


Viruses associated with ill health in wild chimpanzees.
Viral infection is a major cause of ill health in wild chimpanzees (Pan troglodytes), but most evidence to date has come from conspicuous disease outbreaks with high morbidity and mortality. To examine the relationship between viral infection and ill health during periods not associated with disease outbreaks, we conducted a longitudinal study of wild eastern chimpanzees (P. t. schweinfurthii) in the Kanyawara and Ngogo communities of Kibale National Park, Uganda. We collected standardized, observational health data for 4 years and then used metagenomics to characterize gastrointestinal viromes (i.e., all viruses recovered from fecal samples) in individual chimpanzees before and during episodes of clinical disease. We restricted our analyses to viruses thought to infect mammals or primarily associated with mammals, discarding viruses associated with nonmammalian hosts. We found 18 viruses (nine of which were previously identified in this population) from at least five viral families. Viral richness (number of viruses per sample) did not vary by health status. By contrast, total viral load (normalized proportion of sequences mapping to viruses) was significantly higher in ill individuals compared with healthy individuals. Furthermore, when ill, Kanyawara chimpanzees exhibited higher viral loads than Ngogo chimpanzees, and males, but not females, exhibited higher infection rates with certain viruses and higher total viral loads as they aged. Post-hoc analyses, including the use of a machine-learning classification method, indicated that one virus, salivirus (Picornaviridae), was the main contributor to health-related and community-level variation in viral loads. Another virus, chimpanzee stool-associated virus (chisavirus; unclassified Picornavirales), was associated with ill health at Ngogo but not at Kanyawara. Chisavirus, chimpanzee adenovirus (Adenoviridae), and bufavirus (Parvoviridae) were also associated with increased age in males. Associations with sex and age are consistent with the hypothesis that nonlethal viral infections cumulatively reflect or contribute to senescence in long-lived species such as chimpanzees.


CRISPR/Cas9-Assisted Seamless Genome Editing in Lactobacillus plantarum and Its Application in N-Acetylglucosamine Production.
Lactobacillus plantarum is a potential starter and health-promoting probiotic bacterium. Effective, precise, and diverse genome editing of Lactobacillus plantarum without introducing exogenous genes or plasmids is of great importance. In this study, CRISPR/Cas9-assisted double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) recombineering was established in L. plantarum WCFS1 to seamlessly edit the genome, including gene knockouts, insertions, and point mutations. To optimize our editing method, phosphorothioate modification was used to improve the dsDNA insertion, and adenine-specific methyltransferase was used to improve the ssDNA recombination efficiency. These strategies were applied to engineer L. plantarum WCFS1 toward producing N-acetylglucosamine (GlcNAc). nagB was truncated to eliminate the reverse reaction of fructose-6-phosphate (F6P) to glucosamine 6-phosphate (GlcN-6P). Riboswitch replacement and point mutation in glmS1 were introduced to relieve feedback repression. The resulting strain produced 797.3 mg/liter GlcNAc without introducing exogenous genes or plasmids. This strategy may contribute to the available methods for precise and diverse genetic engineering in lactic acid bacteria and boost strain engineering for more applications.IMPORTANCE CRISPR/Cas9-assisted recombineering is restricted in lactic acid bacteria because of the lack of available antibiotics and vectors. In this study, a seamless genome editing method was carried out in Lactobacillus plantarum using CRISPR/Cas9-assisted double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) recombineering, and recombination efficiency was effectively improved by endogenous adenine-specific methyltransferase overexpression. L. plantarum WCFS1 produced 797.3 mg/liter N-acetylglucosamine (GlcNAc) through reinforcement of the GlcNAc pathway, without introducing exogenous genes or plasmids. This seamless editing strategy, combined with the potential exogenous GlcNAc-producing pathway, makes this strain an attractive candidate for industrial use in the future.


Huperzine A attenuates the neurotoxic effect of staurosporine in primary rat cortical neurons.
The effects of huperzine A (HupA) on staurosporine-induced neuronal apoptosis and potential mechanisms were investigated in primary cultured rat cortical neurons. Pretreatment of the cells with HupA (0.1-100 microM) 2 h prior to 0.5 microM staurosporine exposure for 24 h, markedly elevated cell survival. Incubation with HupA at higher dose (1 microM) also reduced the DNA fragmentation caused by staurosporine. This dose of HupA also reduced the upregulation of the pro-apoptotic gene, bax, downregulation of the anti-apoptotic gene, bcl-2, and antagonized the decrease in immunoreactive caspase-3 proenzyme. These results demonstrated that HupA protect neurons against staurosporine-induced apoptosis via regulating the apoptotic related genes and caspase-3 proenzyme.


A novel 51-kDa fragment of the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase formed in the stroma of chloroplasts in dark-induced senescing wheat leaves.
The degradation of large subunit (LSU) of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in wheat (Triticum aestivum L. cv. Yangmai 158) leaves was studied. A novel 51-kDa fragment was detected in leaf crude extracts and in chloroplast lysates from leaves with dark-induced senescence. Further studies showed that the 51-kDa fragment was found in the reaction solution with stroma fraction but not in that with the chloroplast membrane fraction and in the chloroplast lysates from mature wheat leaves. The reaction of producing the 51-kDa fragment was inhibited by 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF), 1,10-phenanthroline and EDTA. The N-terminal sequence analysis indicated that the LSU was cleaved at the peptide bond between Lys-14 and Ala-15. In addition, a 50-kDa fragment of LSU formed obviously at pH 6.0-6.5 was detected in the crude extracts of leaves with dark-induced senescence but was not found in lysates of chloroplasts. The degradation was prevented by AEBSF, leupeptin and transepoxysuccinyl-l-leucylamido (4-guanidino) butane (E-64). The results obtained in this study imply that the appearance of the 51-kDa fragment could be because of the involvement of a new senescence-associated protease that is located in the stroma of chloroplasts in senescing wheat leaves.


Similarity of mRNA phenotypes of morphologically normal macular and peripheral retinal pigment epithelial cells in older human eyes.
To determine the expression profiles of morphologically normal human retinal pigment epithelial (RPE) cells that originate from the macula and periphery. Morphologically normal RPE cells from 15 human globes from donors aged 52 to 82 years old were laser capture microdissected. Total RNA from 5000 cells was SMART amplified, [33]P-labeled, and hybridized to a cDNA array containing 4325 known genes. Expression profiles were analyzed by hierarchical cluster analysis, Prediction Analysis of Microarrays (PAM), and Significance Analysis for Microarrays (SAM). Differentially expressed genes were evaluated further by real time RT-PCR. The overall expression profiles of RPE cells from the macula and periphery were similar. Unsupervised and supervised hierarchical cluster analysis showed that patient genotype was a stronger separating factor than topographical location. SAM analysis identified 11 genes that were underexpressed by macular RPE cells. The expression patterns of these 11 genes were confirmed by real time RT-PCR, with 5 genes reaching statistical significance. Whereas the overall expression profiles were similar between cells from the macula and periphery, subtle differential expression of five genes could contribute to RPE phenotypic differences based on topographic location.


Development of a new epidermal growth factor receptor positron emission tomography imaging agent based on the 3-cyanoquinoline core: synthesis and biological evaluation.
The epidermal growth factor receptor (EGFR/c-ErbB1/HER1) is overexpressed in many cancers including breast, ovarian, endometrial, and non-small cell lung cancer. An EGFR specific imaging agent could facilitate clinical evaluation of primary tumors and/or metastases. To achieve this goal we designed and synthesized a small array of fluorine containing compounds based on a 3-cyanoquinoline core. A lead compound, 16, incorporating 2'-fluoroethyl-1,2,3-triazole was selected for evaluation as a radioligand based on its high affinity for EGFR kinase (IC50=1.81+/-0.18 nM), good cellular potency (IC50=21.97+/-9.06 nM), low lipophilicity and good metabolic stability. 'Click' labeling afforded [18F]16 in 37.0+/-3.6% decay corrected radiochemical yield based on azide [18F]14 and 7% end of synthesis (EOS) yield from aqueous fluoride. Compound [18F]16 was obtained with >99% radiochemical purity in a total synthesis time of 3 h. The compound showed good stability in vivo and a fourfold higher uptake in high EGFR expressing A431 tumor xenografts compared to low EGFR expressing HCT116 tumor xenografts. Furthermore, the radiotracer could be visualized in A431 tumor bearing mice by small animal PET imaging. Compound [18F]16 therefore constitutes a promising radiotracer for further evaluation for imaging of EGFR status.


Postmenopausal hormone therapy: cardiovascular risks.
(1) The WHI study was published in 2002: a randomised double-blind placebo-controlled clinical trial in more than 16 000 women with an average age of 63 years at enrollment. The paper reports data on the long-term adverse effects of combined equine estrogen-progestin hormone replacement therapy, taken for 5 years. (2) On average, a yearly excess of 19 severe adverse events per 10 000 women occurred in the estrogen-progestin group. Relative to the placebo group, there were an extra 8 pulmonary embolisms, 7 coronary events, 8 strokes and 8 cases of invasive breast cancer. In contrast, there were 6 fewer colorectal cancers and 5 fewer hip fractures in the active treatment group. (3) The differences in the frequency of coronary events and venous thromboembolism emerged after the first year of treatment, while the curves for stroke and breast cancer diverged after the second and fifth years, respectively. (4) The overall mortality rate did not differ between the two groups. (5) A placebo-controlled trial of the same hormone combination (HERS trial), given for 4.1 years as secondary prophylaxis against coronary heart disease was published in 1998. The drug was ineffective during the trial, and during unblinded post-trial follow-up of 2 321 women for an average of 2.7 years (HERS II study). (6) The estrogen-progestin combination used in these trials did not reduce the risk of coronary heart disease (in primary or secondary prophylaxis) or the risk of stroke. On the contrary, both risks increased. (7) The increased incidence of deep venous thrombosis and/or pulmonary embolism associated with estrogen-progestin replacement therapy was confirmed in these trials, even among women with no relevant history. (8) The WHI trial also confirmed the increased risk of breast cancer in women on hormone replacement therapy, but did not study its impact on outcome or mortality. (9) The WHI trial confirmed the beneficial impact of estrogen-progestin combination therapy on the risk of osteoporotic fracture. An average of 5 hip fractures were avoided each year per 10 000 women treated (10 versus 15 observed cases per 10 000 women per year), together with 6 symptomatic vertebral fractures (9 versus 15 cases) and 44 osteoporotic fractures (147 versus 191 cases). It is not known whether the benefit observed at the end of the trial persisted after the end of treatment. (10) In practice, the decision to prescribe hormone replacement therapy, and the optimal duration of treatment, must be weighed up according to each individual's risk factors. And the decision to treat or not to treat must be regularly re-assessed. Women must be informed of the potential risks and benefits, and must be monitored. They should also be advised not to use less well assessed treatments such as phytoestrogens, DHEA and tibolone. (11) Health authorities, especially in Europe, must organise comparative trials to assess the benefits and risks of other hormone combinations used by perimenopausal and postmenopausal women.


Apoptotic action of 17beta-estradiol in raloxifene-resistant MCF-7 cells in vitro and in vivo.
Resistance to tamoxifen, a selective estrogen receptor modulator (SERM), involves changes that prevent apoptosis and enhance cell proliferation and survival. Paradoxically, estrogen treatment inhibits the growth of long-term tamoxifen-treated breast tumors. Because of the increasing use of raloxifene, another SERM, to prevent osteoporosis and potentially reduce breast cancer risk, some women will develop raloxifene-resistant breast cancer. We developed a raloxifene-resistant MCF-7 cell model (MCF-7/Ral) and investigated the nature of raloxifene-resistant breast cancer and its response to estradiol. Raloxifene resistance and hormone responsiveness were assessed by proliferation assays and cell cycle analysis in parental MCF-7 and MCF-7/Ral cells. Nuclear factor kappaB (NF-kappaB) activity was investigated with a transient transfection assay. Apoptosis was investigated by annexin V staining, mRNA was measured by real-time polymerase chain reaction, and protein was measured by western blotting. Tumorigenesis was studied by injecting MCF-7 or MCF-7/Ral cells into ovariectomized athymic mice (10 per group) and monitoring tumor size weekly. All statistical tests were two-sided. Basal NF-kappaB activity was higher in MCF-7/Ral cells (1.6 U, 95% confidence interval [CI] = 1.2 to 2.0 U) than in MCF-7 cells (0.8 U, 95% CI = 0.4 to 1.1 U; P =.004). When cultured with 1 microM raloxifene, MCF-7/Ral cells grew statistically significantly (P<.001) faster than MCF-7 cells. Estradiol treatment of MCF-7/Ral cells arrested cells in G(2)/M phase of the cell cycle, decreased NF-kappaB activity (0.2 U, 95% CI = 0.2 to 0.3 U; P<.001), increased expression of Fas protein and mRNA (4.5-fold, 95% CI = 2.8- to 6.3-fold versus 0.5-fold, 95% CI = 0.3- to 0.8-fold for control treatment; P<.001), and induced apoptosis. Treatment with either raloxifene or tamoxifen stimulated MCF-7/Ral tumor growth, suggesting that such tumors were resistant to both drugs. When a 9-week raloxifene or tamoxifen treatment was followed by a 5-week estradiol treatment, estradiol statistically significantly reduced the size of tumors stimulated by raloxifene or tamoxifen (at week 14, P =.004 for raloxifene and P<.001 for tamoxifen). Growth of raloxifene-resistant MCF-7/Ral cells in vitro and in vivo is repressed by estradiol treatment by a mechanism involving G2/M-phase arrest, decreased NF-kappaB activity, and increased Fas expression to induce apoptosis.


Real-time, very-long-baseline interferometry based on the use of a communications satellite.
The Hermes satellite, a joint Canadian-American program, has been used to provide a communication channel between radio telescopes in West Virginia and Ontario, for very-long-baseline interferometry (VLBI). This system makes possible instantaneous correlation of the data as well as a sensitivity substantially better than that of earlier VLBI systems, by virtue of a broader observational bandwidth. With the use of a geostationary communications satellite it is possible to eliminate the tape recorders and the most troublesome part of the postobservational data processing. A further possibility is the development of a phase-coherent interferometer.


International trends in surgical treatment of rectal cancer.
Surgical technique might influence rectal cancer survival, yet international practices for surgical treatment of rectal cancer are poorly described. We performed a cross-sectional survey in a cohort of experienced colorectal surgeons representing 123 centers. Seventy-one percent responded, 70% are from departments performing more than 50 proctectomies annually. More than 50% defined the rectum as "15 cm from the verge." Seventy-two percent perform laparoscopic proctectomy, 80% use oral bowel preparation, 69% perform high ligation of the inferior mesenteric artery, 76% divert stomas as routine for colo-anal anastomosis, and 63% use enhanced recovery protocols. Different practices exist between US and non-US surgeons: 15 cm from the verge to define the rectum (34% vs 59%; P = .03), personally perform laparoscopic resection (82% vs 66%; P = .05), rectal stump washout (36% vs 73%; P = .0001), always drain after surgery (23% vs 42%; P = .03), transanal endoscopic microsurgery for T2N0 in medically unfit patients (39% vs 61%; P = .0001). Wide international variations in rectal cancer management make outcome comparisons challenging, and consensus development should be encouraged.


An extreme value theory approach for the early detection of time clusters. A simulation-based assessment and an illustration to the surveillance of Salmonella.
We propose a new method that could be part of a warning system for the early detection of time clusters applied to public health surveillance data. This method is based on the extreme value theory (EVT). To any new count of a particular infection reported to a surveillance system, we associate a return period that corresponds to the time that we expect to be able to see again such a level. If such a level is reached, an alarm is generated. Although standard EVT is only defined in the context of continuous observations, our approach allows to handle the case of discrete observations occurring in the public health surveillance framework. Moreover, it applies without any assumption on the underlying unknown distribution function. The performance of our method is assessed on an extensive simulation study and is illustrated on real data from Salmonella surveillance in France.


Identification of a genetic variant underlying familial cases of recurrent benign paroxysmal positional vertigo.
Benign paroxysmal positional vertigo (BPPV) is the most common cause of vertigo in humans, yet the molecular etiology is currently unknown. Evidence suggests that genetic factors may play an important role in some cases of idiopathic BPPV, particularly in familial cases, but the responsible genetic variants have not been identified. In this study, we performed whole exome sequencing [including untranslated regions (UTRs)] of 12 families and Sanger sequencing of additional 30 families with recurrent BPPV in Caucasians from the United States (US) Midwest region, to identify the genetic variants responsible for heightened susceptibility to BPPV. Fifty non-BPPV families were included as controls. In silico and experimental analyses of candidate variants show that an insertion variant rs113784532 (frameshift causing truncation) in the neural cadherin gene PCDHGA10 (protocadherin-gamma A10) is an exceedingly strong candidate (p = 1.80x10-4 vs. sample controls; p = 5.85x10-19 vs. ExAC data; p = 4.9x10-3 vs. NHLBI exome data). The mutant protein forms large aggregates in BPPV samples even at young ages, and affected subjects carrying this variant have an earlier onset of the condition than those without [average 44.0±14.0 (n = 16) versus 54.4±16.1 (n = 36) years old, p = 0.054]. In both human and mouse inner ear tissues, PCDHGA10 is expressed in ganglia, hair cells and vestibular transitional epithelia. Fluorescent RNA in situ hybridization using mouse inner ear tissues shows that expression increases with age. In summary, our data show that a variant in the PCDHGA10 gene may be involved in causing or aggravating some familial cases of recurrent idiopathic BPPV.


Evaluation of a tobacco and alcohol use prevention program for Hispanic migrant adolescents: promoting the protective factor of parent-child communication.
Interventions designed to prevent tobacco and alcohol use targeting high-risk adolescents are limited. In addition, few studies have attempted to improve parent-child communication skills as a way of improving and maintaining healthy youth decision-making. A total of 660 Hispanic migrant families participated in a randomized pre-post control group study that was utilized to determine the impact of the intervention on parent-child communication. Both treatment and attention-control groups of youth were exposed to an eight-session culturally sensitive program presented by bilingual/bicultural college students. Parents jointly attended three of the eight sessions and participated in helping their child complete homework assignments supporting the content of each session. The content of the treatment intervention included (1) information about tobacco and alcohol effects, (2) social skills training (i.e., refusal skills), and (3) the specific development of parent-child communication skills to support healthy youth decisions. Significant intervention by household size interactions for both parent and youth perceptions of communication were found indicating that the treatment was effective in increasing communication in families with fewer children. Based on the effect size and the previously established relationship between communication and susceptibility to tobacco and alcohol use, it was determined that the intervention effect could be translated into a future 5 to 10% decrease in susceptibility for these smaller families. A culturally sensitive family-based intervention for migrant Hispanic youth was found to be effective in increasing perceived parent-child communication in families with fewer children. It is expected that increases in this important protective factor will lead to later observed decreases in tobacco and alcohol use.


Phenotypic effect of heterozygous alpha and beta 0-thalassemia interaction.
In this study, we carried out restriction endonuclease mapping in order to characterize the alpha-globin genotype of 10 Sardinian beta 0-thalassemia heterozygotes, all of whom presented with normal red blood cell indices and increased HbA2 levels. In 8 of these subjects, we found the deletion of two alpha-globin genes (-alpha/-alpha), and in the remaining two the deletion of a single alpha-globin gene (-alpha/alpha alpha). In three of these carriers with the (-alpha/-alpha) alpha-globin genotype and in one with the (-alpha/alpha alpha) genotype, we also found the glucose-6-phosphate dehydrogenase (G6PD) defect of the Mediterranean type. On the basis of these findings, we may conclude that the interaction of heterozygous beta 0-thalassemia with alpha-thalassemia, due to the deletion of either one or two alpha-globin genes, may lead to the production of red blood cells with normal indices. The association of the G6PD defect with this thalassemia gene complex may eventually contribute to this effect. We suggest, therefore, that screening programs for heterozygous beta-thalassemia in populations where alpha-thalassemia is also prevalent, should incorporate the determination of HbA2 in the first set of tests.


Oligonucleotides labeled with single fluorophores as sensors for deoxynucleotide triphosphate binding by DNA polymerases.
Oligonucleotides labeled with a single fluorophore (fluorescein or tetramethylrhodamine) have been used previously as fluorogenic substrates for a number of DNA modifying enzymes. Here, it is shown that such molecules can be used as fluorogenic probes to detect the template-dependent binding of deoxynucleotide triphosphates by DNA polymerases. Two polymerases were used in this work: the Klenow fragment of the Escherichia coli DNA polymerase I and the Bacillus stearothermophilus polymerase, Bst. When complexes of these polymerases with dye-labeled hairpin-type oligonucleotides were mixed with various deoxynucleotide triphosphates in the presence of Sr²⁺ as the divalent metal cation, the formation of ternary DNA-polymerase-dNTP complexes was detected by concentration-dependent changes in the fluorescence intensities of the dyes. Fluorescein- and tetramethylrhodamine-labeled probes of identical sequences responded differently to the two polymerases. With Bst polymerase, the fluorescence intensities of all probes increased with the next correct dNTP; with Klenow polymerase, tetramethylrhodamine-labeled probes increased their fluorescence, but the intensity of fluorescein-labeled probes decreased on formation of ternary complexes with the correct incoming nucleotides. The use of Sr²⁺ as the divalent metal ion allowed the formation of catalytically inactive ternary complexes and obviated the need for using 2',3'-dideoxy-terminated oligonucleotides as would have been needed in the case of Mg²⁺ as the metal ion.


The many faces of amyloid beta in Alzheimer's disease.
The 'amyloid cascade hypothesis' links amyloid beta peptide (Abeta) with the pathological process of Alzheimer's disease (AD) and it still awaits universal acceptance. Amyloid precursor protein (APP), through the actions of the gamma-secretase complex, eventually becomes a different Abetaspecies. The various Abeta species have proven to be difficult to investigate under physiological conditions, and the species of Abeta responsible for neurotoxicity has yet to be unequivocally identified. The two important Abeta peptides involved are Abeta(1-40) and Abeta(1-42), and each has been ascribed both toxic and beneficial attributes. The ratio between the two species can be important in AD etiology. Additionally, shorter variants of Abeta peptides such as Abeta(1-8), Abeta(9-16) and Abeta(16) have also been shown to be potential participants in AD pathology. Interestingly, a new 56-kDa Abeta peptide (Abeta*56) disrupts memory when injected into the brains of young rats. Transgenic mice models are complicated by the interplay between various human Abeta types and the mouse Abeta types in the mouse brains. However, the accumulation of Abeta(1-42) in the brains of transgenic C. elegans worms and Drosophila is indeed detrimental. A less investigated aspect of AD is epigenetics, but in time the investigation of the role of epigenetics in AD may add to our understanding of the development of AD.


Autoimmunity in the Lambert-Eaton myasthenic syndrome.
Sera from 64 patients with the Lambert-Eaton myasthenic syndrome (LES) were tested for evidence of autoimmunity to a variety of tissue antigens. One or more organ-specific autoantibodies (thyroid, gastric, and/or skeletal muscle) were found in 29 patients (45%). Of 46 patients without evidence of tumor, autoantibodies were found in 24 (52%), and of 18 patients with tumor, autoantibodies were found in 5 (28%). In an age-matched group of 40 patients with miscellaneous neurologic diseases, 7 (17%) had one or more organ-specific autoantibodies, and in a control group of 47 patients with myasthenia gravis the combined prevalence of thyroid and gastric antibodies was 47%, comparable to that found in LES patients. The prevalence of non-organ-specific autoantibodies (for example, rheumatoid factor and/or antinuclear antibodies) was not significantly elevated in LES. These data clearly justify consideration of LES without tumor as an organ-specific autoimmune disease. If LES with tumor does prove to have an autoimmune basis, the mechanism may involve an antigen common to cholinergic neurons and oat cell carcinoma, both of which are neuroectoderm derivatives.


Phytochemical screening, safety evaluation, anti-inflammatory and analgesic studies of the leaf extracts of Sterculia tragacantha.
Sterculia tragacantha (Sterculiaceae) is used in the treatment of boils, diarrhea, dyspepsia, fever, gonorrhea, snake bite, syphilis, and tapeworm in some West African nations. This study is to investigate its anti-inflammatory and analgesic activities since the plant is being used to treat fever. Fresh leaves of the plant were collected and dried at room temperature and pulverized into powder form and 200 g of this powder was dissolved first in hexane for 72 h and the extract was filtered and the filtrate was concentrated while the substrate was further dissolved in chloroform, ethyl acetate and methanol at different times and similar procedure adopted as for the hexane. The organic solvents were used based on order of increasing polarity. Graded concentrations of the solvent extracts were prepared and used for the study. Pilot toxicity test was carried out to determine safety dose using hematology and serum chemistry as indices of toxicity. Thereafter anti-inflammatory and analgesic studies were conducted using standard tests such as carrageenan, histamine-induced-edema, tail flick test and acetic writhing test. Phytochemical screening of the plant was also conducted. Phytochemical screening of the powdered material showed that alkaloid, flavonoid and reducing sugar were present while tannin, cardiac glycosides, saponins and anthraquinones were absent. Pilot toxicity test using aqueous extract at 100 mg/mL concentration showed that no mortality was recorded although the animals that received 3,000 mg/kg dose exhibited slight dullness after 48 h. No significant changes were also observed for the packed cell volume, hemoglobin, white blood cell counts, platelet counts, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, blood urea nitrogen, albumin, globulin except for the 200 and 3,000 mg/kg doses that caused significant increase in the level of total protein. The n-hexane, chloroform, ethyl acetate, and methanol extracts of S. tragacantha and indomethacin produced significant (p<0.05) inhibition of paw edema compared with the control using histamine and carrageenan methods of paw edema induction. There was significant (p<0.05) reduction in writhing movements at 100 mg/kg and 200 mg/kg of n-hexane, chloroform, and ethyl acetate leaf extracts of S. tragacantha and indomethacin (10 mg/kg) when compared to the control. This effect using tail flick test was not as effective when compared to the writhing test. The different leaf extracts of S. tragacantha exhibited anti-inflammatory and analgesic properties and they are also safe for medicinal use.


Novelty-evoked elevations of nucleus accumbens dopamine: dependence on impulse flow from the ventral subiculum and glutamatergic neurotransmission in the ventral tegmental area.
In vivo microdialysis in freely moving rats was used to monitor novelty-evoked elevations in extracellular dopamine in the nucleus accumbens septi (NAS) and to examine the role of the ventral subiculum of the hippocampus and glutamatergic transmission in the ventral tegmental area (VTA) on these elevations. Exposure to novel stimuli evoked investigatory activity and increased nucleus accumbens dopamine. Unilateral injections of the sodium channel blocker tetrodotoxin (0.16 ng/0.5 microL) into the ventral subiculum ipsilateral to the dialysed NAS abolished novelty-evoked elevations in dopamine. Injections of tetrodotoxin into the contralateral VS did not prevent novelty-evoked elevations in nucleus accumbens dopamine. Unilateral perfusion (via microdialysis) of the ionotropic glutamate receptor antagonists kynurenic acid (1 mM) into the ipsilateral but not the contralateral VTA blocked novelty-evoked elevations in nucleus accumbens dopamine. Neither unilateral injections of tetrodotoxin nor unilateral perfusion of kynurenic acid disrupted investigatory behaviour. These data indicate that phasic elevations in nucleus accumbens dopamine evoked by exposure to unconditioned novel stimuli are dependent on impulse flow from the hippocampus and glutamatergic transmission in the VTA.


Roles of thromboxane A2 and 5-hydroxytryptamine in endotoxin-induced digital vasoconstriction in horses.
To evaluate the roles of 5-hydroxytryptamine (5-HT), thromboxane A2 (TxA2), and platelet-activating factor (PAF) in endotoxin-induced digital hypoperfusion in horses. 6 healthy adult Thoroughbreds. Horses were treated with IV administration of saline (0.9% NaCl) solution (control treatment) or the 5-HT 1B/D selective antagonist, GR55562 (0.3 mg/kg), prior to tryptamine infusion (1.6 microg/kg/min for 30 minutes) to establish an effective GR55562 dose. In a crossover study, horses were treated with IV administration of saline solution (control treatment), aspirin (4 mg/kg, 2 hours or 4 days before lipopolysaccharide [LPS] infusion), GR55562 (0.3 mg/kg), the PAF antagonist WEB2086 (3 mg/kg), or aspirin plus GR55562 prior to LPS infusion (30 ng/kg for 30 minutes). Digital blood flow was measured by use of Doppler ultrasonography. Concomitant measurements of hoof wall and coronary band surface temperatures were made. Serial blood samples were collected and plasma 5-HT and TxA2 concentrations determined. GR55562 abolished tryptamine-induced digital hypoperfusion. Neither WEB2086 nor GR55562 affected LPS-induced alterations in digital perfusion or plasma mediator concentrations. Aspirin given 2 hours before LPS administration abolished the increase in plasma TxA2 concentration and significantly attenuated LPS-induced digital hypoperfusion. Aspirin given 4 days before LPS significantly attenuated the increase in plasma TxA2 concentration and digital hypothermia. Aspirin plus GR55562 had a greater effect on LPS-induced digital hypothermia than aspirin alone. Thromboxane A2 and 5-HT played a role in mediating LPS-induced digital hypoperfusion in horses. Platelet-activating factor appeared unimportant in mediating LPS-induced 5-HT or TxA2 release or digital hypoperfusion.


Multicomponent Biomarker Approach Improves the Accuracy of Diagnostic Biomarkers for Psoriasis Vulgaris.
Accurate biomarker-based diagnosis of psoriasis vulgaris has remained a challenge; no reliable disease-specific biomarkers have yet been identified. There are several different chronic inflammatory skin diseases that can present similar clinical and dermoscopy features to psoriasis vulgaris, making accurate diagnosis more difficult. Both literature-based and data-driven selection of biomarker was conducted to select candidates for a multicomponent biomarker for psoriasis vulgaris. Support vector machine-based classification models were trained using gene expression data from locally recruited patients and validated on 7 public datasets, which included gene expression data of other inflammatory skin diseases in addition to psoriasis vulgaris. The resulting accuracy of the best classification model based on the expression levels of 4 genes (IL36G, CCL27, NOS2 and C10orf99) was 96.4%, outperforming classification based on other marker gene combinations, which were more affected by variability in gene expression profiles between different datasets and patient groups. This approach has the potential to fill the void of clinically applicable diagnostic biomarkers for psoriasis vulgaris and other inflammatory skin diseases.


A comprehensive neuropsychological mapping battery for functional magnetic resonance imaging.
Existing batteries for FMRI do not precisely meet the criteria for comprehensive mapping of cognitive functions within minimum data acquisition times using standard scanners and head coils. The goal was to develop a battery of neuropsychological paradigms for FMRI that can also be used in other brain imaging techniques and behavioural research. Participants were 61 healthy, young adult volunteers (48 females and 13 males, mean age: 22.25 ± 3.39 years) from the university community. The battery included 8 paradigms for basic (visual, auditory, sensory-motor, emotional arousal) and complex (language, working memory, inhibition/interference control, learning) cognitive functions. Imaging was performed using standard functional imaging capabilities (1.5-T MR scanner, standard head coil). Structural and functional data series were analysed using Brain Voyager QX2.9 and Statistical Parametric Mapping-8. For basic processes, activation centres for individuals were within a distance of 3-11 mm of the group centres of the target regions and for complex cognitive processes, between 7 mm and 15 mm. Based on fixed-effect and random-effects analyses, the distance between the activation centres was 0-4 mm. There was spatial variability between individual cases; however, as shown by the distances between the centres found with fixed-effect and random-effects analyses, the coordinates for individual cases can be used to represent those of the group. The findings show that the neuropsychological brain mapping battery described here can be used in basic science studies that investigate the relationship of the brain to the mind and also as functional localiser in clinical studies for diagnosis, follow-up and pre-surgical mapping.


Radiation Treatment Time and Overall Survival in Locally Advanced Non-small Cell Lung Cancer.
Prolonged radiation treatment (RT) time (RTT) has been associated with worse survival in several malignancies. The present study investigated whether delays during RT are associated with overall survival (OS) in non-small cell lung cancer (NSCLC). The National Cancer Database was queried for patients with stage III NSCLC who had received definitive concurrent chemotherapy and fractionated RT to standard doses (59.4-70.0 Gy) and fractionation from 2004 to 2013. The RTT was classified as standard or prolonged for each treatment regimen according to the radiation dose and number of fractions. Cox proportional hazards models were used to evaluate the association between the following factors and OS: RTT, RT fractionation, demographic and pathologic factors, and chemotherapeutic agents. Of 14,154 patients, the RTT was prolonged in 6262 (44.2%). Factors associated with prolonged RTT included female sex (odds ratio [OR] 1.21, P<.0001), black race (OR 1.20, P=.001), nonprivate health insurance (OR 1.30, P<.0001), and lower income (<$63,000 annually, OR 1.20, P<.0001). The median OS was significantly worse for patients with prolonged RTT than that for those with standard RTT (18.6 vs 22.7 months, P<.0001). Furthermore, the OS worsened with each cumulative interval of delay (standard RTT vs prolonged 1-2 days, 20.5 months, P=.009; prolonged 3-5 days, 17.9 months, P<.0001; prolonged 6-9 days, 17.7 months, P<.0001; prolonged >9 days, 17.1 months, P<.0001). On multivariable analysis, prolonged RTT was independently associated with inferior OS (hazard ratio 1.21, P<.0001). Prolonged RTT as a continuous variable was also significantly associated with worse OS (hazard ratio 1.001, P=.0007). Delays during RT appear to negatively affect survival for patients with locally advanced NSCLC. We have detailed the demographic and socioeconomic barriers influencing prolonged RTT as a method to address the health disparities in this regard. Cumulative interruptions of RT should be minimized.


Assessment of endogenous and therapeutic arteriogenesis by contrast ultrasound molecular imaging of integrin expression.
We hypothesized that molecular imaging with contrast-enhanced ultrasound (CEU) and microbubbles targeted to endothelial integrins could be used to noninvasively assess early angiogenic responses to ischemia and growth factor therapy. Hindlimb ischemia was produced in 48 rats by ligation of an iliac artery. Half of the animals received intramuscular sustained-release fibroblast growth factor-2 (FGF-2). Immediately after ligation and at subsequent intervals from 4 to 28 days, blood flow and oxygen tension in the proximal adductor muscles were measured by CEU perfusion imaging and phosphor quenching, respectively. Targeted CEU imaging of alpha(v)- and alpha5beta1-integrin expression was performed with microbubbles bearing the disintegrin echistatin. Iliac artery ligation produced a 65% to 70% reduction in blood flow and oxygen tension. In untreated ischemic muscle, muscle flow and oxygen tension partially recovered by days 14 to 28. In these animals, signal from integrin-targeted microbubbles was intense and peaked before flow increase (days 4 to 7). In comparison to untreated animals, FGF-2-treated muscle had a greater rate and extent of blood flow recovery and greater signal intensity from integrin-targeted microbubbles, which peaked before maximal recovery of flow. On immunohistology, arteriolar but not capillary density increased in the ischemic limb after ligation, the rate and degree of which were greater in FGF-2-treated rats. Immunofluorescence demonstrated intense staining for alpha(v) in arterioles, the temporal course of which correlated with targeted imaging. Targeted CEU can be used to assess endogenous and therapeutic arteriogenesis before recovery of tissue perfusion. These results suggest that molecular imaging of integrin expression may be useful for evaluating proangiogenic therapies.


Surgical management of acute cholecystitis at a tertiary care center in the modern era.
The advent of laparoscopy has changed the paradigm of surgical training and care delivery for the treatment of patients with acute cholecystitis (AC). Retrospective data collection and analysis. Hospital admissions with a primary diagnosis of AC at a tertiary care center from January 1, 2002, to January 1, 2007. During the study period, 923 patients were admitted with a primary diagnosis of AC. One hundred fourteen patients were excluded from the study because of missing data, medical management, incomplete operative notes or documents, or metastatic gastrointestinal cancer. Patient demographics, preoperative morbidity, procedures (medical and surgical), and postoperative outcomes were statistically analyzed using chi(2) test, t test, and analysis of variance. Eight hundred nine patients (87.6%) with a primary diagnosis of AC underwent surgery by 44 surgeons. Procedures included 663 laparoscopic cholecystectomies (LCs) (82.0%), 9 open cholecystectomies (1.1%), 51 conversions from LC to open cholecystectomy (6.3%), and 86 cholecystostomy tube placements (10.6%). During the study period, cholecystostomy tube placements increased, while open cholecystectomies and conversions from LC to open cholecystectomy decreased (P < .05). Laparoscopic cholecystectomy was associated with significantly better outcomes, including shorter postsurgical stay (2.2 vs 6.3 days for other modalities) and fewer complications (8.5% vs 17.0%). Based on 5-year results from a tertiary care center, LC was performed with a low conversion rate to open surgery and was associated with decreased morbidity and mortality compared with other surgical modalities to treat AC. Our data confirm the benefits and widespread use of LC in the modern era, reflecting changes in the training paradigm and learning curve for laparoscopy.