Design and evaluation of peptide nucleic acid probes for specific identification of Candida albicans.
Candida albicans is an important cause of systemic fungal infections, and rapid diagnostics for identifying and differentiating C. albicans from other Candida species are critical for the timely application of appropriate antimicrobial therapy, improved patient outcomes, and pharmaceutical cost savings. In this work, two 28S rRNA-directed peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) probes, P-Ca726 (targeting a novel region of the ribosome) and P-CalB2208 (targeting a previously reported region), were evaluated. Hybridization conditions were optimized by using both fluorescence microscopy (FM) and flow cytometry (FCM), and probes were screened for specificity and discriminative ability against a panel of C. albicans and various nontarget Candida spp. The performance of these PNA probes was compared quantitatively against that of DNA probes or DNA probe/helper combinations directed against the same target regions. Ratiometric analyses of FCM results indicated that both the hybridization quality and yield of the PNA probes were higher than those of the DNA probes. In FCM-based comparisons of the PNA probes, P-Ca726 was found to be highly specific, showing 2.5- to 5.5-fold-higher discriminatory power for C. albicans than P-CalB2208. The use of formamide further improved the performance of the new probe. Our results reinforce the significant practical and diagnostic advantages of PNA probes over their DNA counterparts for FISH and indicate that P-Ca726 may be used advantageously for the rapid and specific identification of C. albicans in clinical and related applications, especially when combined with FCM.


Acetate Synthesis from H(2) plus CO(2) by Termite Gut Microbes.
Gut microbiota from Reticulitermes flavipes termites catalyzed an H(2)-dependent total synthesis of acetate from CO(2). Rates of H(2)-CO(2) acetogenesis in vitro were 1.11 +/- 0.37 mumol of acetate g (fresh weight) h (equivalent to 4.44 +/- 1.47 nmol termite h) and could account for approximately 1/3 of all the acetate produced during the hindgut fermentation. Formate was also produced from H(2) + CO(2), as were small amounts of propionate, butyrate, and lactate-succinate. However, H(2)-CO(2) formicogenesis seemed largely unrelated to acetogenesis and was believed not to be a significant reaction in situ. Little or no CH(4) was formed from H(2) + CO(2) or from acetate. H(2)-CO(2) acetogenesis was inhibited by O(2), KCN, CHCl(3), and iodopropane and could be abolished by prefeeding R. flavipes with antibacterial drugs. By contrast, prefeeding R. flavipes with starch resulted in almost complete defaunation but had little effect on H(2)-CO(2) acetogenesis, suggesting that bacteria were the acetogenic agents in the gut. H(2)-CO(2) acetogenesis was also observed with gut microbiota from Prorhinotermes simplex, Zootermopsis angusticollis, Nasutitermes costalis, and N. nigriceps; from the wood-eating cockroach Cryptocercus punctulatus; and from the American cockroach Periplaneta americana. Pure cultures of H(2)-CO(2)-acetogenic bacteria were isolated from N. nigriceps, and a preliminary account of their morphological and physiological properties is presented. Results indicate that in termites, CO(2) reduction to acetate, rather than to CH(4), represents the main electron sink reaction of the hindgut fermentation and can provide the insects with a significant fraction (ca. 1/3) of their principal oxidizable energy source, acetate.


Studies in healthy volunteers can demonstrate bronchodilator activity of orally administered drugs.
A panel of healthy volunteers was screened by bronchial histamine provocation. The response used was the flow rate achieved during a partial forced expiratory manoeuvre at a lung volume 30% of the vital capacity above the residual volume. Those subjects who were relatively sensitive to the bronchoconstrictor effects of inhaled histamine took part in a reproducibility study. Those with consistent responses took part in a further study in which the bronchodilator, and bronchoconstriction-protective, effects of orally administered chlorpheniramine, terfenadine and salbutamol were investigated. The technique was able to demonstrate the efficacy of these drugs in non-patient volunteers. Thus it seems to be a suitable method for the early evaluation, in man, of certain drugs with an anti-asthma therapeutic potential. It also provides useful information on the dose and duration of action of compounds prior to the first efficacy trials in patients.


Predicting climate-driven regime shifts versus rebound potential in coral reefs.
Climate-induced coral bleaching is among the greatest current threats to coral reefs, causing widespread loss of live coral cover. Conditions under which reefs bounce back from bleaching events or shift from coral to algal dominance are unknown, making it difficult to predict and plan for differing reef responses under climate change. Here we document and predict long-term reef responses to a major climate-induced coral bleaching event that caused unprecedented region-wide mortality of Indo-Pacific corals. Following loss of >90% live coral cover, 12 of 21 reefs recovered towards pre-disturbance live coral states, while nine reefs underwent regime shifts to fleshy macroalgae. Functional diversity of associated reef fish communities shifted substantially following bleaching, returning towards pre-disturbance structure on recovering reefs, while becoming progressively altered on regime shifting reefs. We identified threshold values for a range of factors that accurately predicted ecosystem response to the bleaching event. Recovery was favoured when reefs were structurally complex and in deeper water, when density of juvenile corals and herbivorous fishes was relatively high and when nutrient loads were low. Whether reefs were inside no-take marine reserves had no bearing on ecosystem trajectory. Although conditions governing regime shift or recovery dynamics were diverse, pre-disturbance quantification of simple factors such as structural complexity and water depth accurately predicted ecosystem trajectories. These findings foreshadow the likely divergent but predictable outcomes for reef ecosystems in response to climate change, thus guiding improved management and adaptation.


LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing.
Little is known about the protein constituents of autophagosome membranes in mammalian cells. Here we demonstrate that the rat microtubule-associated protein 1 light chain 3 (LC3), a homologue of Apg8p essential for autophagy in yeast, is associated to the autophagosome membranes after processing. Two forms of LC3, called LC3-I and -II, were produced post-translationally in various cells. LC3-I is cytosolic, whereas LC3-II is membrane bound. The autophagic vacuole fraction prepared from starved rat liver was enriched with LC3-II. Immunoelectron microscopy on LC3 revealed specific labelling of autophagosome membranes in addition to the cytoplasmic labelling. LC3-II was present both inside and outside of autophagosomes. Mutational analyses suggest that LC3-I is formed by the removal of the C-terminal 22 amino acids from newly synthesized LC3, followed by the conversion of a fraction of LC3-I into LC3-II. The amount of LC3-II is correlated with the extent of autophagosome formation. LC3-II is the first mammalian protein identified that specifically associates with autophagosome membranes.


Hyperosmolar dextrose injection for recalcitrant Osgood-Schlatter disease.
To examine the potential of dextrose injection versus lidocaine injection versus supervised usual care to reduce sport alteration and sport-related symptoms in adolescent athletes with Osgood-Schlatter disease. Girls aged 9 to 15 and boys aged 10 to 17 were randomly assigned to either therapist-supervised usual care or double-blind injection of 1% lidocaine solution with or without 12.5% dextrose. Injections were administered monthly for 3 months. All subjects were then offered dextrose injections monthly as needed. Unaltered sport (Nirschl Pain Phase Scale < 4) and asymptomatic sport (Nirschl Pain Phase Scale = 0) were the threshold goals. Sixty-five knees in 54 athletes were treated. Compared with usual care at 3 months, unaltered sport was more common in both dextrose-treated (21 of 21 vs 13 of 22; P = .001) and lidocaine-treated (20 of 22 vs 13 of 22; P = .034) knees, and asymptomatic sport was more frequent in dextrose-treated knees than either lidocaine-treated (14 of 21 vs 5 of 22; P = .006) or usual-care-treated (14 of 21 vs 3 of 22; P < .001) knees. At 1 year, asymptomatic sport was more common in dextrose-treated knees than knees treated with only lidocaine (32 of 38 vs 6 of 13; P = .024) or only usual care (32 of 38 vs 2 of 14; P < .0001). Our results suggest superior symptom-reduction efficacy of injection therapy over usual care in the treatment of Osgood-Schlatter disease in adolescents. A significant component of the effect seems to be associated with the dextrose component of a dextrose/lidocaine solution. Dextrose injection over the apophysis and patellar tendon origin was safe and well tolerated and resulted in more rapid and frequent achievement of unaltered sport and asymptomatic sport than usual care.


Atrial natriuretic peptide attenuates the development of pulmonary hypertension in rats adapted to chronic hypoxia.
To test the hypothesis that chronic infusion of atrial natriuretic peptide (ANP) instituted before hypoxic exposure attenuates the development of pulmonary hypertension in hypoxia adapted rats, ANP (0.2 and 1.0 microgram/h) or vehicle was administered intravenously via osmotic minipump for 4 wk beginning before exposure to 10% O2 or to room air. Low dose ANP increased plasma ANP levels by only 60% of vehicle controls after 4 wk and significantly decreased mean pulmonary arterial pressure (MPAP) (P less than 0.01), the ratio of right ventricular weight to body weight (RV/BW) (P less than 0.01), and the wall thickness of small (50-100 microns) pulmonary arteries (P = 0.01) in hypoxia-adapted rats. ANP did not alter any of these parameters in air-control rats. High dose ANP increased plasma ANP levels by 230% of control and produced greater reductions in MPAP (P less than 0.001) and RV/BW) (P less than 0.05), but not in pulmonary arterial wall thickness, than the low dose. Neither dose of ANP altered mean systemic arterial pressure in either hypoxic or normoxic rats. The data demonstrate that chronic infusion of exogenous ANP at a dose that does not affect MPAP or RV weight in air-control rats attenuates the development of pulmonary hypertension and RV enlargement in rats adapted to chronic hypoxia.


Traffic safety: Rights and obligations.
Morbidity and mortality due to road traffic injuries (RTI) is one of the few public health problems where society and decision makers still accept death and disability on such a large scale as inevitable. Discussion only revolves around the number of deaths and injuries we are willing to accept. The partial departure from this mode of thinking is 'Vision Zero' for road safety that was adopted by the Swedish parliament in 1997. The long-term objective is that no one shall be killed or seriously injured in traffic and that the design, function and use of the transport system shall be adapted to the standards this requires. In this article we try to understand the concept of RTI as a public health problem and why that understanding has led to the introduction of Vison Zero and then sporadic attempts to establish road safety as a fundamental human right. We provide some details surrounding these events, the reasons for their limited success, and suggest ways in how we might move forward in establishing a place for rights and obligations to ensure road safety in reality. Some of the ways forward include: (a) Every policy, law or safety standard (concerning roads, vehicles or traffic management) established by the state to be accompanied by a justification for the same by including systematic reviews of the scientific evidence used for the decision and the expected safety benefits in numerical estimates. (b) Manufacturers of vehicles and other road-based technologies to explicitly state the quality and limits of the safety features embedded in their technologies. (c) International agencies dealing with road safety (state and non-state) to examine all sources of systematic reviews of road safety interventions and use them to justify the policies they pursue. They should also make it explicit that they will fund road safety activity by non-government organisations only if they promote interventions justified by scientific evidence.


Distinct neural specializations for learning to read words and name objects.
Understanding the neural systems that underpin reading acquisition is key if neuroscientific findings are to inform educational practice. We provide a unique window into these systems by teaching 19 adults to read 24 novel words written in unfamiliar letters and to name 24 novel objects while in an MRI scanner. Behavioral performance on trained items was equivalent for the two stimulus types. However, componential letter-sound associations were extracted when learning to read, as shown by correct reading of untrained words, whereas object-name associations were holistic and arbitrary. Activity in bilateral anterior fusiform gyri was greater during object name learning than learning to read, and ROI analyses indicated that left mid-fusiform activity was predictive of success in object name learning but not in learning to read. In contrast, activity in bilateral parietal cortices was predictive of success for both stimulus types but was greater during learning and recall of written word pronunciations relative to object names. We argue that mid-to-anterior fusiform gyri preferentially process whole items and contribute to learning their spoken form associations, processes that are required for skilled reading. In contrast, parietal cortices preferentially process componential visual-verbal mappings, a process that is crucial for early reading development.


Longitudinal associations between conduct problems and depressive symptoms among girls and boys with early conduct problems.
Youth with conduct problems (CP) may experience high rates of depressive symptoms (DS). However, little is known about the direction of the longitudinal associations between CP and DS in this specific population. Although girls with CP appear at greater risk than boys for presenting comorbid depression, empirical research on gender differences in these associations is even sparser. The current study used autoregressive latent trajectory models to compare four perspectives with hypotheses regarding the longitudinal associations between CP and DS, while taking into account the evolution of both problems. We also examined gender differences in the longitudinal associations. A total of 345 children (40.6 % female) presenting with a high level of CP in early elementary school (mean age at study inception = 8.52; SD = .94) were evaluated annually over a four-year period (5 measurement time points). The results revealed that CP and DS were quite stable over time. Moreover, CP and DS showed strong covariation at each measurement time point, but only one significant positive cross-lagged association between the two processes, indicating that higher levels of DS at time 3 were associated with higher levels of CP 1 year later. No differences were observed in the longitudinal associations between CP and DS in boys and girls. Given the comorbidity and stability of CP and DS, these findings suggest that DS should be systematically evaluated among children with early clinically significant CP, and treatment plans should include interventions aimed at both CP and DS among children who present with both types of problems.


STUDIES ON THE OXIDATION AND REDUCTION OF IMMUNOLOGICAL SUBSTANCES : VIII. THE ANTIGENIC PROPERTIES OF HEMOLYTICALLY ACTIVE AND HEMOLYTICALLY INACTIVE MODIFICATIONS OF PNEUMOCOCCUS HEMOTOXIN.
THE FOLLOWING MODIFICATIONS OF THE ANTIGEN (PNEUMOCOCCUS HEMOTOXIN) WERE STUDIED: (1) the hemolytically active (reduced) substance; (2) the hemolytically inactive, reversible oxidation product; (3) the inactive irreversible products formed by treatment With high concentrations of H(2)O(2); (4) the inactive products formed by heat. The antibody-invoking property of the reversibly oxidized form seemed to be identical with that of the original, hemolytically active or reduced form; neither of the other two hemolytically inactive products invoked antibody production. The same modifications of the antigen which exhibited the antibody-invoking property in vivo possessed the antibody-combining property in vitro; and the modifications which lacked the one property also lacked the other. Evidence is presented that the groups of the hemotoxin molecule in which the true antigenic properties are resident are not necessarily altered by processes which inactivate the groupings responsible for the toxic (hemolytic) action of the original antigen. The lack of antigenic properties on the part of the other two hemolytically inactive modifications is evidence that the treatment employed to alter the toxic property of the molecule must be properly chosen to avoid profound changes which affect the antigenically effective groupings. From an immunological point of view, the reversibility of the antigenically effective oxidation product of pneumococcus hemotoxin is important as an index that the loss of toxicity (hemolysis) was accomplished without a profound change in the molecule. The theoretical significance of the antigenicity of non-toxic modifications of toxic antigens is discussed.


Reduced Lateral Inhibition Impairs Olfactory Computations and Behaviors in a Drosophila Model of Fragile X Syndrome.
Fragile X syndrome (FXS) patients present neuronal alterations that lead to severe intellectual disability, but the underlying neuronal circuit mechanisms are poorly understood. An emerging hypothesis postulates that reduced GABAergic inhibition of excitatory neurons is a key component in the pathophysiology of FXS. Here, we directly test this idea in a FXS Drosophila model. We show that FXS flies exhibit strongly impaired olfactory behaviors. In line with this, olfactory representations are less odor specific due to broader response tuning of excitatory projection neurons. We find that impaired inhibitory interactions underlie reduced specificity in olfactory computations. Finally, we show that defective lateral inhibition across projection neurons is caused by weaker inhibition from GABAergic interneurons. We provide direct evidence that deficient inhibition impairs sensory computations and behavior in an in vivo model of FXS. Together with evidence of impaired inhibition in autism and Rett syndrome, these findings suggest a potentially general mechanism for intellectual disability.


Capsaicin-sensitive nerves mediate esophageal mucosal protection.
The esophageal mucosa is exposed to damaging agents both by ingestion and reflux. Using our in vivo rabbit model of esophagitis, we have observed that acute luminal exposure (within 1 to 5 minutes) to potentially harmful agents, such as acid, bile, or ethanol, induces a rapid increase in mucosal blood flow; whereas prolonged exposure (10 to 60 minutes) results in mucosal injury and ablation of blood flow. We have also shown that capsaicin-sensitive mucosal afferent nerves can modulate esophageal blood flow. These findings led us to hypothesize that the reactive increase in blood flow induced by luminal agents represents a mechanism of protection mediated by capsaicin-sensitive nerves. The objective of these experiments was to determine if luminal capsaicin, a specific probe for visceral afferent nerves, could both preserve mucosal blood flow and protect against ethanol injury. Rabbits were subjected to luminal instillation of 50% ethanol with or without 1% capsaicin. Blood flow was measured with microspheres at baseline and after 2 and 10 minutes. Rabbits exposed only to ethanol developed severe mucosal injury coincident with near ablation of mucosal blood flow. In contrast, rabbits exposed to ethanol with capsaicin showed protection of the epithelium with a sixfold increase in mucosal blood flow. We conclude that capsaicin-sensitive nerves in the esophagus are local effectors of mucosal protection by virtue of preserving blood flow.


Plication procedures-excisional and incisional corporoplasty and imbrication for Peyronie's disease.
Plication procedures for the correction of Peyronie's disease (PD) curvature are management options for PD patients. There are basically three types of procedures: excisional corporoplasty, incisional corporoplasty, and plication-only. This review is a compilation of English literature, peer-reviewed, published articles addressing these types of operations for Peyronie's curvature correction, not congenital curvature. According to the urology literature, this surgical type was initially used for correction of curvature associated with hypospadias repair or congenital penile curvature. The procedures also, for the most part, historically became an alternative for plaque excision and graft repair, because of the difficulty with such repairs and the often-resultant erectile dysfunction (ED). A brief section traces some of the origins of these various repairs, followed by a brief section on the selection criteria for these types of surgery for the patient with PD penile curvature. We also review the significant articles in which the three types were presented with modifications. Finally, several articles that compare the various surgical repairs are summarized in the order that they appear in the literature. These types of surgery have become a mainstay for the surgical correction of penile curvature due to PD.


Maternal vaccination against pertussis: a systematic review of the recent literature.
This study is conducted to summarise and present the current knowledge on antenatal vaccination against pertussis with regard to national recommendations, coverage, immunogenicity, safety and effectiveness of the current available vaccines. A systematic review of the literature in English was undertaken from January 2011 to May 2016 with searches in four databases. The review conformed to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. 47 studies fulfilled the inclusion criteria. Antenatal vaccination against pertussis induces high antibody concentrations in pregnant women, which are efficiently transferred transplacentally to the fetus and protect newborns when they are most vulnerable to pertussis. This strategy has been demonstrated to be safe, with no evidence of adverse pregnancy, birth or neonatal outcomes. Several countries have already introduced antenatal pertussis vaccination into their national immunisation programme with varying vaccination coverage influenced by various factors. Barriers to achieving high immunisation rates could be improved through better education of the public and healthcare professionals. There is now an increasing body of evidence to support the safety, immunogenicity and effectiveness of antenatal vaccination to reduce the morbidity and mortality associated with pertussis in neonates and young infants before they receive their primary immunisations. Narrowing the gap between scientific evidence and public health policies is critical in order to protect the most vulnerable as quickly as possible. The lessons learnt have important implications for implementation of new vaccines into the antenatal immunisation programme.


Gene therapy for achromatopsia.
The present review summarizes the current status of achromatopsia (ACHM) gene therapy-related research activities and provides an outlook for their clinical application. ACHM is an inherited eye disease characterized by a congenital absence of cone photoreceptor function. As a consequence, ACHM is associated with strongly impaired daylight vision, photophobia, nystagmus and a lack of color discrimination. Currently, six genes have been linked to ACHM. Up to 80% of the patients carry mutations in the genes CNGA3 and CNGB3 encoding the two subunits of the cone cyclic nucleotide-gated channel. Various animal models of the disease have been established and their characterization has helped to increase our understanding of the pathophysiology associated with ACHM. With the advent of adeno-associated virus vectors as valuable gene delivery tools for retinal photoreceptors, a number of promising gene supplementation therapy programs have been initiated. In recent years, huge progress has been made towards bringing a curative treatment for ACHM into clinics. The first clinical trials are ongoing or will be launched soon and are expected to contribute important data on the safety and efficacy of ACHM gene supplementation therapy.


Effect of isouronium/guanidinium substitution on the efficacy of a series of novel anti-cancer agents.
Considering our hypothesis that the guanidinium moiety in the protein kinase type III inhibitor 1 interacts with a phosphate of ATP within the hinge region, the nature of the interactions established between a model isouronium and the phosphate groups of ATP was computationally analysed indicating that an isouronium derivative of 1 will interact in a similar manner with ATP. Thus, a number of compounds were prepared to assess the effect of the guanidinium/isouronium substitution on cancer cell growth; additionally, the molecular shortening and conformational change induced by replacing the di-substituted guanidine-linker of 1 by an amide was explored. The effect of these compounds on cell viability was tested in human leukaemia, breast cancer and cervical cancer cell lines and the resulting IC50 values were compared with those of the lead compound 1. Replacement of the di-substituted guanidine-linker by an amide results in the loss of cytotoxicity; however, substitution of the mono-substituted guanidinium by an isouronium cation seems to be beneficial for cell growth inhibition. Additionally, the effect of these compounds on the MAPK/ERK pathway was studied by means of Western blotting and the results indicate that the isouronium derivative 2 decreases the levels of phosphorylated, and thus activated, ERK (pERK) both in leukaemia and breast cancer cells, whereas lead compound 1 only shows an effect on pERK levels in breast cancer cells. This confirms that both compounds could interfere with the MAPK/ERK pathway although other targets cannot be ruled out.


SU-E-J-90: 2D/3D Registration Using KV-MV Image Pairs for Higher Accuracy Image Guided Radiotherapy.
In this work, we investigate the impact of using paired portal mega-voltage (MV) and kilo-voltage (kV) images, on 2D/3D registration accuracy with the purpose of improving tumor motion tracking during radiotherapy. Tumor motion tracking is important as motion remains one of the biggest sources of uncertainty in dose application. 2D/3D registration is successfully used in online tumor motion tracking, nevertheless, one limitation of this technique is the inability to resolve movement along the imaging beam axis using only one projection image. Our evaluation consisted in comparing the accuracy of registration using different 2D image combinations: only one 2D image (1-kV), one kV and one MV image (1kV-1MV) and two kV images (2-kV). For each of the image combinations we evaluated the registration results using 250 starting points as initial displacements from the gold standard. We measured the final mean target registration error (mTRE) and the success rate for each registration. Each of the combinations was evaluated using four different merit functions. When using the MI merit function (a popular choice for this application) the RMS mTRE drops from 6.4 mm when using only one image to 2.1 mm when using image pairs. The success rate increases from 62% to 99.6%. A similar trend was observed for all four merit functions. Typically, the results are slightly better with 2-kV images than with 1kV-1MV. We evaluated the impact of using different image combinations on accuracy of 2D/3D registration for tumor motion monitoring. Our results show that using a kV-MV image pair, leads to improved results as motion can be accurately resolved in six degrees of freedom. Given the possibility to acquire these two images simultaneously, this is not only very workflow efficient but is also shown to be a good approach to improve registration accuracy.


Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein-Protein Interactions.
Optogenetics enables the spatio-temporally precise control of cell and animal behavior. Many optogenetic tools are driven by light-controlled protein-protein interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs). Applying light-controlled PPIs to new target proteins is challenging because it is difficult to predict which of the many available LSDs, if any, will yield robust light regulation. As a consequence, fusion protein libraries need to be prepared and tested, but methods and platforms to facilitate this process are currently not available. Here, we developed a genetic engineering strategy and vector library for the rapid generation of light-controlled PPIs. The strategy permits fusing a target protein to multiple LSDs efficiently and in two orientations. The public and expandable library contains 29 vectors with blue, green or red light-responsive LSDs, many of which have been previously applied ex vivo and in vivo. We demonstrate the versatility of the approach and the necessity for sampling LSDs by generating light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a new resource for optical regulation of a broad range of target proteins in cell and developmental biology.


Advances in endoscopic imaging of colorectal neoplasia.
Colon cancer screening is arguably the most important activity performed by gastroenterologists. Recent decreases in rates of death from colorectal cancer indicate that screening methods such as colonoscopy have a positive impact. There is still room for improvement, however, particularly in prevention of right-sided colon cancer. Practice issues, such as making colonoscopy more comfortable, safer, and less costly, are keys to continued success in cancer prevention. Colonoscopy techniques, technologies, and quality control measures have advanced to improve detection, classification, and removal of early neoplasias. In particular, slow, careful inspection of the colon by gastroenterologists who have been trained in lesion recognition has improved rates of detection of polypoid and flat neoplasias. Image enhancement methods such as chromoendoscopy have greatly improved neoplasia detection in patients with chronic colitis, but are not widely used because they are perceived as inconvenient. More convenient methods, such as "digital" chromoendoscopy, show promise but have had mixed results. Ultra-high magnification systems, including optical magnification and confocal endomicroscopy, can be used during the colonoscopy examination to evaluate small polyps, allowing physicians to make immediate diagnoses and decisions about whether to remove polyps. In patients with inflammatory bowel disease, improved imaging techniques could eliminate the needs for analysis of randomly selected biopsy samples and resection of all (neoplastic and non-neoplastic) polyps. It is important to maintain high standards of quality for colonoscopy examination, detection, and removal of high-risk lesions, as well as to make colon cancer screening more widely accepted and affordable for the entire at-risk population.


Improved quality of management of eclampsia patients through criteria based audit at Muhimbili National Hospital, Dar es Salaam, Tanzania. Bridging the quality gap.
Criteria-based audits (CBA) have been used to improve clinical management in developed countries, but have only recently been introduced in the developing world. This study discusses the use of a CBA to improve quality of care among eclampsia patients admitted at a University teaching hospital in Dar es Salaam Tanzania. The prevalence of eclampsia in MNH is high (≈6%) with the majority of cases arriving after start of convulsions. In 2004-2005 the case-fatality rate in eclampsia was 5.1% of all pregnant women admitted for delivery (MNH obstetric data base). A criteria-based audit (CBA) was used to evaluate the quality of care for eclamptic mothers admitted at Muhimbili National Hospital (MNH), Dar es Salaam, Tanzania after implementation of recommendations of a previous audit. A CBA of eclampsia cases was conducted at MNH. Management practices were evaluated using evidence-based criteria for appropriate care. The Ministry of Health (MOH) guidelines, local management guidelines, the WHO manual supplemented by the WHO Reproductive Health Library, standard textbooks, the Cochrane database and reviews in peer reviewed journals were adopted. At the initial audit in 2006, 389 case notes were assessed and compared with the standards, gaps were identified, recommendations made followed by implementation. A re-audit of 88 cases was conducted in 2009 and compared with the initial audit. There was significant improvement in quality of patient management and outcome between the initial and re-audit: Review of management plan by senior staff (76% vs. 99%; P=0.001), urine for albumin test (61% vs. 99%; P=0.001), proper use of partogram to monitor labour (75% vs. 95%; P=0.003), treatment with steroids for lung maturity (2.0% vs. 24%; P=0.001), Caesarean section within 2 hours of decision (33% vs. 61%; P=0.005), full blood count (28% vs. 93%; P=0.001), serum urea and creatinine (44% vs. 86%; P=0.001), liver enzymes (4.0% vs. 86%; P=0.001), and specialist review within 2 hours of admission (25% vs. 39%; P=0.018). However, there was no significant change in terms of delivery within 24 hours of admission (69% vs. 63%; P=0.33). There was significant reduction of maternal deaths (7.7% vs. 0%; P=0.001). CBA is applicable in low resource setting and can help to improve quality of care in obstetrics including management of pre-eclampsia and eclampsia.


TGFbeta receptor II gene deletion in leucocytes prevents cerebral vasculitis in bacterial meningitis.
In bacterial meningitis, chemokines lead to recruitment of polymorphonuclear leucocytes (PMN) into the CNS. At the site of infection in the subarachnoid space, PMN release reactive oxygen species, reactive nitrogen intermediates (RNI) and interleukin-1beta (IL-1beta). Although these immune factors assist in clearance of bacteria, they also result in neuronal injury associated with meningitis. Transforming growth factor beta (TGFbeta) is a potent deactivator of PMN and macrophages since TGFbeta suppresses the production of ROI, RNI and IL-1. Here, we report that the deletion of the TGFbeta receptor II gene in PMN enhances PMN recruitment into the CNS of mice with Streptococcus pneumoniae meningitis. This was associated with more efficient clearance of bacteria, and almost complete prevention of intracerebral necrotizing vasculitis. Differences in PMN in the CNS of infected control mice and mice lacking TGFbeta receptor II were not explained by altered expression of chemokines acting on PMN. Instead, TGFbeta was found to impair the expression of L (leucocyte)-selectin on PMN from control mice but not from mice lacking TGFbeta receptor II. L-selectin is known to be essential for PMN recruitment in bacterial meningitis. We conclude that defective TGFbeta signalling in PMN is beneficial in bacterial meningitis by ameliorating migration of PMN and bacterial clearance.


The Ews-ERG fusion protein can initiate neoplasia from lineage-committed haematopoietic cells.
The EWS-ERG fusion protein is found in human sarcomas with the chromosomal translocation t(21;22)(q22;q12), where the translocation is considered to be an initiating event in sarcoma formation within uncommitted mesenchymal cells, probably long-lived progenitors capable of self renewal. The fusion protein may not therefore have an oncogenic capability beyond these progenitors. To assess whether EWS-ERG can be a tumour initiator in cells other than mesenchymal cells, we have analysed Ews-ERG fusion protein function in a cellular environment not typical of that found in human cancers, namely, committed lymphoid cells. We have used Ews-ERG invertor mice having an inverted ERG cDNA cassette flanked by loxP sites knocked in the Ews intron 8, crossed with mice expressing Cre recombinase under the control of the Rag1 gene to give conditional, lymphoid-specific expression of the fusion protein. Clonal T cell neoplasias arose in these mice. This conditional Ews gene fusion model of tumourigenesis shows that Ews-ERG can cause haematopoietic tumours and the precursor cells are committed cells. Thus, Ews-ERG can function in cells that do not have to be pluripotent progenitors or mesenchymal cells.


Prognostic value of serum concentration of heart-type fatty acid-binding protein relative to cardiac troponin T on admission in the early hours of acute coronary syndrome.
Heart-type fatty acid-binding protein (H-FABP) is proposed as an early biomarker for acute myocardial infarction (AMI), but its prognostic value is unclear in acute coronary syndrome (ACS). We evaluated the prognostic value of the H-FABP concentration relative to cardiac troponin T (cTnT) in the early hours of ACS. Serum concentrations of H-FABP and cTnT were measured on admission in 328 consecutive patients hospitalized for ACS within 6 h after the onset of chest pain [AMI, 241 (73.5%) patients; ST-segment elevation myocardial infarction, 154 (47.0%) patients; and emergent coronary angiography within 24 h after admission, 287 (87.5%) patients]. Cardiac events, which were defined as cardiac death or subsequent nonfatal AMI, were monitored for 6 months after admission. During the 6-month follow-up period, there were 25 cardiac events, including 15 cardiac deaths and 10 subsequent nonfatal AMIs. Stepwise multivariate analyses including clinical, electrocardiographic, and biochemical variables revealed that increased H-FABP (above the median of 9.8 microg/L), but not increased cTnT (above the median of 0.02 microg/L), was independently associated with cardiac events in all patients [relative risk (RR) = 8.96; P = 0.0004], the subgroup of patients with ST-segment elevation myocardial infarction (RR = 11.3; P = 0.02), and the subgroup of patients with unstable angina and non-ST-segment elevation myocardial infarction (RR = 8.31; P = 0.007). The area under the ROC curve was higher for H-FABP than for cTnT (0.711 vs 0.578; P = 0.08), suggesting that H-FABP concentrations have a greater predictive capacity for cardiac events than cTnT. Serum H-FABP is a potential independent predictor of cardiac events within 6 months of patient admission and may provide prognostic information superior to cTnT in the early hours of ACS.


Tree species diversity of natural mixed stands in eastern Black Sea and western Mediterranean region of Turkey.
Differences in free species diversity of natural mixed stands were compared between the eastern Black sea region (EBSR) and western Mediterranean region (WMR) of Turkey, to clarify the effects of differences in forest structure, focusing on the tree species occurring in each. Species diversity with special reference to stand structure, of natural mixed stands was quantified by Shannon-Wiener index (H'), equitability index (J'), and species richness index (R). All species diversity indices were significantly higher in EBSR than in WMR. According to the coefficients of homogeneity (CH), stands in EBSR are generally have uneven-aged stand structure, but in WMR even-aged stand structure is more common. Uneven-aged stands have more tree species diversity than even-aged forests due to complex vertical forest structure and species composition. According to Pearson's coefficients, species diversity indices and richness are closely related the average stand diameter (Ds) in uneven-aged stands of EBSR, but average stand diameter (Ds), age (A), and stand density (SDI) are the most important stand parameters in even-aged stands of WMR in Turkey.


Elongation and migration of single DNA molecules in microchannels using oscillatory shear flows.
Much of modern biology relies on the strategic manipulation of molecules for creating ordered arrays prior to high throughput molecular analysis. Normally, DNA arrays involve deposition on surfaces, or confinement in nanochannels; however, we show that microfluidic devices can present stretched molecules within a controlled flow in ways complementing surface modalities, or extreme confinement conditions. Here we utilize pressure-driven oscillatory shear flows generated in microchannels as a new way of stretching DNA molecules for imaging "arrays" of individual DNA molecules. Fluid shear effects both stretch DNA molecules and cause them to migrate away from the walls becoming focused in the centerline of a channel. We show experimental findings confirming simulations using Brownian dynamics accounting for hydrodynamic interactions between molecules and channel-flow boundary conditions. Our findings characterize DNA elongation and migration phenomena as a function of molecular size, shear rate, oscillatory frequency with comparisons to computer simulation studies.


Development of a DNA-based microarray for the detection of zoonotic pathogens in rodent species.
The demand for diagnostic tools that allow simultaneous screening of samples for multiple pathogens is increasing because they overcome the limitations of other methods, which can only screen for a single or a few pathogens at a time. Microarrays offer the advantages of being capable to test a large number of samples simultaneously, screening for multiple pathogen types per sample and having comparable sensitivity to existing methods such as PCR. Array design is often considered the most important process in any microarray experiment and can be the deciding factor in the success of a study. There are currently no microarrays for simultaneous detection of rodent-borne pathogens. The aim of this report is to explicate the design, development and evaluation of a microarray platform for use as a screening tool that combines ease of use and rapid identification of a number of rodent-borne pathogens of zoonotic importance. Nucleic acid was amplified by multiplex biotinylation PCR prior to hybridisation onto microarrays. The array sensitivity was comparable to standard PCR, though less sensitive than real-time PCR. The array presented here is a prototype microarray identification system for zoonotic pathogens that can infect rodent species.


The diagnostic role of 18-fluorodeoxyglucocose-positron emission tomography/computed tomography in occult bacteremia searching underlying primary disease.
18-Fluoro-2-deoxy-D-glucose(FDG) is a structural analog of 2-deoxyglucose and accumulates in malignant tissues but also at sites of infection and inflammation. For this reason, FDG PET or PET/CT has great advantage in understanding of underlying pathology in assessment of FUO (Fever of unknown origin). However, till today, there are limited studies about the role of FDG PET or PET/CT in evaluation of FUO. Conventional diagnostic methods are still not adequate to reveal underlying reason in approximately 50% of patients with FUO especially in cases presenting with diagnostic challenges i.e. involvement of two or more organ systems with seemingly no correlation. We report a case of two years old Indian female child who presented with fever of one month duration, CT and MRI reported nonspecific findings. She underwent Whole body 18 FDG PET/CT for further evaluation, which revealed FDG avid rim lesion with central photopenic defect suspicious of pyogenic abscess in high parietal cortex along with bilateral lung nodules. This confirmed the diagnosis of a brain abscess secondary to pulmonary infection. We emphasize the utility of 18 FDG PET/CT as imaging modality, highlight the diagnostic difficulties using current serological and radiological measures, and propose managing FUO with 18 FDG PET/CT in cases empirically prior to more invasive measures.


3D patient-specific numerical modeling of the soft palate considering adhesion from the tongue.
Collapse of the soft palate in the upper airway contributes to obstructive sleeping apnea (OSA). In this study, we investigate the influence of the adhesion from the tongue on the soft palate global response. This is achieved using a cohesive zone finite element approach. A traction-separation law is determined to describe the adhesion effect from the surface tension of the lining liquid between the soft palate and the tongue. According to pull-off experimental tests of human lining liquid from the oral surface of the soft palate, the corresponding cohesive properties, including the critical normal traction stress and the failure separation displacement, are obtained. The 3D patient-specific soft palate geometry is accounted for, based on one specific patient's computed tomography (CT) images. The calculation results show that influence of the adhesion from the tongue surface on the global response of the soft palate depends on the length ratio between the cohesive length and the soft palate length. When the length of the cohesive zone is smaller than half of the soft palate length, the adhesion's influence is negligible. When the adhesion length is larger than 70 percent of soft palate length, the adhesion force contributes to preventing the soft palate from collapsing towards to the pharynx wall, i.e. the closing pressure is more negative than in the no adhesion case. These results may provide useful information to the clinical treatment of OSA patients.


SERS-active metal-dielectric nanostructures integrated in microfluidic devices for label-free quantitative detection of miRNA.
In this work, SERS-based microfluidic PDMS chips integrating silver-coated porous silicon membranes were used for the detection and quantitation of microRNAs (miRNAs), which consist of short regulatory non-coding RNA sequences typically over- or under-expressed in connection with several diseases such as oncogenesis. In detail, metal-dielectric nanostructures which provide noticeable Raman enhancements were functionalized according to a biological protocol, adapted and optimized from an enzyme-linked immunosorbent assay (ELISA), for the detection of miR-222. Two sets of experiments based on different approaches were designed and performed, yielding a critical comparison. In the first one, the labelled target miRNA is revealed through hybridization to a complementary thiolated DNA probe, immobilized on the silver nanoparticles. In the second one, the probe is halved into shorter strands (half1 and half2) that interact with the complementary miRNA in two steps of hybridization. Such an approach, taking advantage of the Raman labelling of half2, provides a label-free analysis of the target. After suitable optimisation of the procedures, two calibration curves allowing quantitative measurements were obtained and compared on the basis of the SERS maps acquired on the samples loaded with several miRNA concentrations. The selectivity of the two-step assay was confirmed by the detection of target miR-222 mixed with different synthetic oligos, simulating the hybridization interference coming from similar sequences in real biological samples. Finally, that protocol was applied to the analysis of miR-222 in cellular extracts using an optofluidic multichamber biosensor, confirming the potentialities of SERS-based microfluidics for early-cancer diagnosis.


The workflow of single-cell expression profiling using quantitative real-time PCR.
Biological material is heterogeneous and when exposed to stimuli the various cells present respond differently. Much of the complexity can be eliminated by disintegrating the sample, studying the cells one by one. Single-cell profiling reveals responses that go unnoticed when classical samples are studied. New cell types and cell subtypes may be found and relevant pathways and expression networks can be identified. The most powerful technique for single-cell expression profiling is currently quantitative reverse transcription real-time PCR (RT-qPCR). A robust RT-qPCR workflow for highly sensitive and specific measurements in high-throughput and a reasonable degree of multiplexing has been developed for targeting mRNAs, but also microRNAs, non-coding RNAs and most recently also proteins. We review the current state of the art of single-cell expression profiling and present also the improvements and developments expected in the next 5 years.


Investigation of ambulatory blood pressure monitoring data editing criteria.
Ambulatory blood pressure monitoring (ABPM) allows multiple measurements of BP over a 24h period but often produces a proportion of measurements which are distant from the mean and unlikely to be genuine. The software which accompanies the SpaceLabs 90207 monitor requires criteria to be set which excludes outliers (autoediting) but there is no published data evaluating at what levels these autoediting criteria should be set. This study set out to ascertain whether outlying BP measurements represent true blood pressures or machine error, the determinants of machine error and appropriate autoedit criteria. Twenty subjects, both normotensive and hypertensive, were studied by ABPM with a blinded observer recording BP using a standard mercury sphygmomanometer (SMS) connected via a T-tube during three activity phases: sitting with cuffed arm still, walking but keeping arm still during measurement and sitting but gently moving the arm during measurement. The results show that a high ABPM/SMS discrepancy is associated with arm movement. After studying different autoediting criteria which aimed to eliminate the maximum number of conflicting readings while excluding the minimum number of unconflicting readings, an autoediting threshold of between 1.75 and 2 s.d.s from the mean based separately on sleep and awake measurements is suggested.


Junior doctors' views about careers in academic medicine.
To investigate junior doctors' views about careers in academic medicine. Postal questionnaire survey. National Health Service in England. Doctors in university posts at specialist registrar level, Medical Research Council and Wellcome Trust training fellows, and specialist registrars in National Health Service posts. Incentives to pursue an academic career which respondents rated as strong related to the challenge of research and the intellectual environment of research units. The strongest disincentives were perceived difficulties in obtaining research grants and uncertainty regarding pay parity with National Health Service colleagues. Medical Research Council and Wellcome fellows had much more protected research time than other academic doctors but were less satisfied with their clinical training. Academic doctors who were not fellows reported spending less than half their time on research and the great majority agreed that their research suffers when there is pressure on the service side. The job content of academic posts should be kept under regular review to ensure that clinical service pressures do not inappropriately erode research time while also ensuring that postholders have adequate clinical training. Training programmes need flexibility to accommodate the needs of clinical academics in their progress through higher specialist training.


Nuclear actin aggregation is a hallmark of anti-synthetase syndrome-induced dysimmune myopathy.
To analyze antisynthetase syndrome-associated myositis by modern myopathologic methods and to define its place in the spectrum of idiopathic inflammatory myopathies (IIMs). Skeletal muscle biopsies from antisynthetase syndrome-associated myositis and other IIMs from different institutions worldwide were analyzed by histopathology, quantitative PCR, and electron microscopy. Myonuclear actin filament inclusions were identified as a unique morphologic hallmark of antisynthetase syndrome-associated myositis. Nuclear actin inclusions were never found in dermatomyositis, polymyositis, sporadic inclusion body myositis, autoimmune necrotizing myopathy associated with signal recognition particle or 3-hydroxy-3-methylglutaryl-coenzyme A reductase autoantibodies, or nonspecific myositis associated with other systemic diseases, harboring myositis-associated autoantibodies, and presenting myofiber necrosis. We show that molecules involved in actin filament formation and actin shuttling mechanisms are altered in antisynthetase syndrome, and may thus be involved in pathologic myonuclear actin aggregation. In addition, we have identified a typical topographic distribution of necrotic myofibers predominantly located at the periphery of muscle fascicles accompanied by inflammation and destruction of the perimysial connective tissue. Antisynthetase syndrome-associated myositis is characterized by distinctive myonuclear actin filament inclusions, including rod formations and a typical necrotizing perimysial myositis. This supports the hypothesis that antisynthetase syndrome-associated myositis is unique and should not be grouped among dermatomyositis, polymyositis, sporadic inclusion body myositis, necrotizing autoimmune myositis, or nonspecific myositis. This study provides Class II evidence that for patients with IIMs, the presence of myonuclear actin filament inclusions accurately identifies patients with antisynthetase syndrome-associated myositis (sensitivity 81%, specificity 100%).


The spatial distribution of health vulnerability to heat waves in Guangdong Province, China.
International literature has illustrated that the health impacts of heat waves vary according to differences in the spatial variability of high temperatures and the social and economic characteristics of populations and communities. However, to date there have been few studies that quantitatively assess the health vulnerability to heat waves in China. To assess the spatial distribution of health vulnerability to heat waves in Guangdong Province, China. A vulnerability framework including dimensions of exposure, sensitivity, and adaptive capacity was employed. The last two dimensions were called social vulnerability. An indicator pool was proposed with reference to relevant literatures, local context provided by relevant local stakeholder experts, and data availability. An analytic hierarchy process (AHP) and a principal component analysis were used to determine the weight of indicators. A multiplicative vulnerability index (VI) was constructed for each district/county of Guangdong province, China. A total of 13 items (two for exposure, six for sensitivity, and five for adaptive capacity) were proposed to assess vulnerability. The results of an AHP revealed that the average VI in Guangdong Province was 0.26 with the highest in the Lianzhou and Liannan counties of Qingyuan (VI=0.50) and the lowest in the Yantian district of Shenzhen (VI=0.08). Vulnerability was gradiently distributed with higher levels in northern inland regions and lower levels in southern coastal regions. In the principal component analysis, three components were isolated from the 11 social vulnerability indicators. The estimated vulnerability had a similar distribution pattern with that estimated by AHP (Intraclass correlation coefficient (ICC)=0.98, p<0.01). Health vulnerability to heat waves in Guangdong Province had a distinct spatial distribution, with higher levels in northern inland regions than that in the southern coastal regions.


Molecular cloning, over expression and characterization of thermoalkalophilic esterases isolated from Geobacillus sp.
Due to potential use for variety of biotechnological applications, genes encoding thermoalkalophilic esterase from three different Geobacillus strains isolated from thermal environmental samples in Balçova (Agamemnon) geothermal site were cloned and respective proteins were expressed in Escherichia coli (E.coli) and characterized in detail. Three esterases (Est1, Est2, Est3) were cloned directly by PCR amplification using consensus degenerate primers from genomic DNA of the strains Est1, Est2 and Est3 which were from mud, reinjection water and uncontrolled thermal leak, respectively. The genes contained an open reading frame (ORF) consisting of 741 bp for Est1 and Est2, which encoded 246 amino acids and ORF of Est3 was 729 bp encoded 242 amino acids. The esterase genes were expressed in E. coli and purified using His-Select HF nickel affinity gel. The molecular mass of the recombinant enzyme for each esterase was approximately 27.5 kDa. The three esterases showed high specific activity toward short chain p-NP esters. Recombinant Est1, Est2, Est3 have exhibited similar activity and the highest esterase activity of 1,100 U/mg with p-nitrophenyl acetate (pNPC(2)) as substrate was observed with Est1. All three esterase were most active around 65°C and pH 9.5-10.0. The effect of organic solvents, several metal ions, inhibitors and detergents on enzyme activity for purified Est1, Est2, Est3 were determined separately and compared.


Right ventricular normal measurements: time to index?
Despite the common practice of indexing left ventricular dimensions to body surface area, there remains a lack of indexed normal right ventricular (RV) two-dimensional caliper measurements. Variations in ranges for normal RV dimensions have been shown to exist, and indexing RV dimensions according to body surface area may help reduce this and provide a standardization useful for clinical practice. The aim of this study was to prospectively establish both absolute and indexed normal dimensions for the right ventricle using standardized positions in a multiethnic population. Furthermore, the effects of both gender and ethnicity on both the absolute and indexed results were also evaluated. Two hundred five healthy volunteers from four ethnic backgrounds (Indian, Chinese, Malay, and European) were prospectively enrolled and underwent two-dimensional echocardiography according to a set protocol. Ten measurements were made in conjunction with previous research. Intraobserver and interobserver and test-retest variability was assessed using coefficients of variation and intraclass correlation coefficients. Male absolute results exceeded female absolute results in 90% of measurements (P = .003). European absolute results (male and female) were significantly larger in up to eight of 10 measurements (P = .01). When indexed, female results became significantly larger (P < .001) than male results. Indexing was able to reduce the number of statistical differences between male ethnic groups. Measurements showed good levels of intraobserver and interobserver variability for apical and short-axis measurements. Gender and body surface area play an important part in the determination of normal RV reference ranges, whereas ethnicity has little influence. Results using the suggested RV markers for these measurements showed good repeatability.


The practical challenges of evaluating a blanket emergency feeding programme in northern Kenya.
A blanket supplementary feeding programme for young children was implemented for four months in five northern districts of Kenya from January 2010 because of fears of food insecurity exacerbated by drought. An attempt to evaluate the impact of the food on children's anthropometric status was put in place in three districts. The main aim of the analysis was to assess the quality of the data on the cohort of children studied in the evaluation and to propose methods by which it could be improved to evaluate future blanket feeding programmes. Data on the name, age, sex, weight and height of a systematic sample of children recruited at 61 food distribution sites were collected at the first, second and third rounds and again at an extra, fifth food distribution, offered only to the evaluation subjects. Of the 3,544 children enrolled, 483 (13.63%) did not collect a fifth ration. Of the 2,640 children who were considered by their name to be the same at the first and fifth food distribution (13% were different), data on only 902 children (34.17%) were considered acceptable based on their age (an arbitrary ±3 months different) and their length or height (between >-1 or ≤4 cm different) at the two instances they were seen. Data on nearly two thirds of children were of questionable quality. The main reasons for the poor quality data were inconsistencies in estimating age or because caretakers may have brought different children. Recommendations are made about how to improve data quality including ensuring that entry to a blanket feeding programme is clearly based on height, not age, to avoid misreporting age; careful identification of subjects at all contacts; and using well-trained, specialist evaluation staff.


The Impact of Initial COVID-19 Episode Inflammation Among Adults on Mortality Within 12 Months Post-hospital Discharge.
Inflammation in the initial COVID-19 episode may be associated with post-recovery mortality. The goal of this study was to determine the relationship between systemic inflammation in COVID-19 hospitalized adults and mortality after recovery from COVID-19. An analysis of electronic health records (EHR) for patients from 1 January, 2020 through 31 December, 2021 was performed for a cohort of COVID-19 positive hospitalized adult patients. 1,207 patients were followed for 12 months post COVID-19 episode at one health system. 12-month risk of mortality associated with inflammation, C-reactive protein (CRP), was assessed in Cox regressions adjusted for age, sex, race and comorbidities. Analyses evaluated whether steroids prescribed upon discharge were associated with later mortality. Elevated CRP was associated other indicators of severity of the COVID-19 hospitalization including, supplemental oxygen and intravenous dexamethasone. Elevated CRP was associated with an increased mortality risk after recovery from COVID-19. This effect was present for both unadjusted (HR = 1.60; 95% CI 1.18, 2.17) and adjusted analyses (HR = 1.61; 95% CI 1.19, 2.20) when CRP was split into high and low groups at the median. Oral steroid prescriptions at discharge were found to be associated with a lower risk of death post-discharge (adjusted HR = 0.49; 95% CI 0.33, 0.74). Hyperinflammation present with severe COVID-19 is associated with an increased mortality risk after hospital discharge. Although suggestive, treatment with anti-inflammatory medications like steroids upon hospital discharge is associated with a decreased post-acute COVID-19 mortality risk.


The synergistic effect of hierarchical assemblies of siRNA and chemotherapeutic drugs co-delivered into hepatic cancer cells.
Diblock copolymers (PEI-PCL) of poly(ε-caprolactone) (PCL) and linear poly(ethylene imine) (PEI) were synthesized and assembled to biodegradable nano-carriers for co-delivery of BCL-2 siRNA and doxorubicin (DOX). Folic acid as a tumor-targeting ligand was conjugated to the polyanion, poly(ethylene glycol)-block-poly(glutamic acid) (FA-PEG-PGA). Driven by the electrostatic interaction, FA-PEG-PGA was coated onto the surface of the cationic PEI-PCL nanoparticles pre-loaded with siRNA and DOX, potentiating a ligand-directed delivery to human hepatic cancer cells Bel-7402. At certain N/P and C/N ratios (N/P: PEI-PCL nitrogen to siRNA phosphate; C/N: FA-PEG-PGA carboxyl to PEI-PCL amine), the nanoparticles exhibited not only high transfection efficiency but also ideally controlled release of drug. Compared to non-specific delivery, the folate-targeted delivery of BCL-2 siRNA resulted in more significant gene suppression at both the BCL-2 mRNA and protein expression levels, inducing cancer cell apoptosis and improving the therapeutic efficacy of the co-administered DOX. Herein we demonstrated that co-loading siRNA and small molecular drug in a multifunctional hierarchical nano-assembly enabled simultaneously delivering siRNA and drug into the same cancer cells, yielding synergistic effect of RNA interference and chemotherapy in cancer.


Vaccine-induced rabies infection in rural dogs in Anambra State, Nigeria.
Six cases of apparent vaccination failures in rural dogs given modified live virus, chicken embryo origin, low egg passage, Flury-type vaccine, were investigated. Four cases were dumb rabies while two were furious forms of rabies. Human exposure (six people) was attributed to the furious form only. Four brain smears from dogs which died of dumb rabies were positive for Negri bodies while two brain smears obtained from dogs which died of the furious form of rabies were negative. All the brain samples were positive when subjected to a mouse inoculation test. Contact with other dogs was eliminated. Furthermore, the time lapse between vaccination and development of clinical signs (incubation period) varied from 4 to 14 days. The 'break' in vaccination was therefore linked with the vaccine used. Dumb rabies was observed more than the furious form of rabies in this study, with considerable potential health hazards.


Entacapone and tolcapone, two catechol O-methyltransferase inhibitors, block fibril formation of alpha-synuclein and beta-amyloid and protect against amyloid-induced toxicity.
Parkinson disease (PD) is the second most common neurodegenerative disorder after Alzheimer disease (AD). There is considerable consensus that the increased production and/or aggregation of alpha-synuclein (alpha-syn) plays a central role in the pathogenesis of PD and related synucleinopathies. Current therapeutic strategies for treating PD offer mainly transient symptomatic relief and aim at the restitution of dopamine levels to counterbalance the loss of dopaminergic neurons. Therefore, the identification and development of drug-like molecules that block alpha-synuclein aggregation and prevent the loss of dopaminergic neurons are desperately needed to treat or slow the progression of PD. Here, we show that entacapone and tolcapone are potent inhibitors of alpha-syn and beta-amyloid (Abeta) oligomerization and fibrillogenesis, and they also protect against extracellular toxicity induced by the aggregation of both proteins. Comparison of the anti-aggregation properties of entacapone and tolcapone with the effect of five other catechol-containing compounds, dopamine, pyrogallol, gallic acid, caffeic acid, and quercetin on the oligomerization and fibrillization of alpha-syn and Abeta, demonstrate that the catechol moiety is essential for the anti-amyloidogenic activity. Our findings present the first characterization of the anti-amyloidogenic properties of tolcapone and entacapone against both alpha-synuclein and Abeta42 and highlight the potential of this class of nitro-catechol compounds as anti-amyloidogenic agents. Their inhibitory properties, mode of action, and structural properties suggest that they constitute promising lead compounds for further optimization.


[Consensus on acute ischemic stroke].
Stroke is the third cause of death and the first cause of disability in Argentina. Ischemic events constitute 80% of cases. It requires the implementation of systematized protocols that allow reducing the time of care, morbidity and mortality. Specialists from nine medical societies related to the care of patients with cerebrovascular disease participated in the consensus. A separate agenda was agreed upon in chapters and for the writing of them, work groups were formed with members of different medical specialties. The level of recommendation was discussed and agreed upon for each topic based on the best clinical evidence available for each of them. An adaptation to the local scope of the recommendations was made when it was considered necessary.The American Heart Association system was used to draft the recommendations and their level of evidence. The correction and editing were done by five external reviewers, who did not participate in the writing and with extensive experience in vascular pathology. Once the preliminary document was finalized, a general meeting was held with all the members of the working groups and the reviewers to reach final recommendations. The consensus covers the management of ischemic stroke in the pre-hospital phase, initial evaluation in the emergency center, recanalization therapies (thrombolysis and/ or mechanical thrombectomy), decompressive craniectomy, neuroimaging and clinical care in the hospital.


Combination of microdialysis and glucose sensor for continuous on line measurement of the subcutaneous glucose concentration: theory and practical application.
The microdialysis technique can be used to get dialysates of the subcutaneous tissue, which can be continuously measured by an amperometric glucose sensor. In order to get further insight into the microdialysis procedure, we used a steady-state theory for microdialysis to predict the recovery of glucose in the dialysate and compared the results to experimental data obtained by a combination of the microdialysis technique with continuous amperometric glucose sensing. The recovery of glucose obtained in vitro for two different microdialysis probes was close to the theoretical predictions. When quantifying the predictions of the model with regard to the spatial concentration profile in the subcutaneous tissue, it appeared, that the presence of the microdialysis probe depressed the concentration of glucose for 0.2 mm from the probe surface. In a 24 hour in vivo experiment, there were less fluctuations in the sensor signal when the patient was lying in bed compared to the time, when the patient could move freely. In conclusion, the combination of microdialysis and glucose sensor seems to be a promising approach to a continuously functioning glucose sensing system. However, the microdialysis procedure itself disturbs the surrounding of the probe leading to a concentration gradient of glucose. This might explain some differences between the course of blood glucose and the course of subcutaneous glucose, measured by the combination of microdialysis and an amperometric glucose sensor. Further developments of such systems should aim at implanting microdialysis devices which have a minimal influence upon the tissue metabolism.


Invaded Invaders: Infection of Invasive Brown Treesnakes on Guam by an Exotic Larval Cestode with a Life Cycle Comprised of Non-Native Hosts.
Multiple host introductions to the same non-native environment have the potential to complete life cycles of parasites incidentally transported with them. Our goal was to identify a recently detected parasitic flatworm in the invasive Brown Treesnake (Boiga irregularis) on the remote Pacific island of Guam. We considered possible factors influencing parasite transmission, and tested for correlations between infection status and potential indicators of host fitness. We used genetic data from the parasite and information about the native ranges of other possible non-native hosts to hypothesize how it arrived on Guam and how its life cycle may be currently supported. We identified the parasite by comparing larval morphology and mtDNA sequences with other Pseudophyllid tapeworms. We assessed probability of infection in individual snakes using logistic regression and examined different factors influencing presence of parasites in hosts. We identified the parasite as the pseudophyllid cestode Spirometra erinaceieuropaei, with all sampled worms from multiple snakes sharing a single mtDNA haplotype. Infection appears to be limited to the only freshwater watershed on the island, where infection prevalence was high (77.5%). Larger snakes had a higher probability of being infected, consistent with the chronic nature of such infections. While infection status was positively correlated with body condition, infected snakes tended to have lower intra-peritoneal fat body mass, potentially indicating a negative effect on energy stores. We discovered that B. irregularis inhabiting a small area of forested habitat in a freshwater watershed on Guam are often infected by a novel parasite of Asian origin. While further work is needed, this species of Spirometra, itself a non-native species, likely depends on a suite of recently introduced hosts from different parts of the world to complete the life cycle. This baseline study provides little evidence of any effects on host fitness, but additional data are needed to more thoroughly explore the consequences of infection in this invasive snake population.


Insights from human genetic studies of lung and organ fibrosis.
Genetic investigations of fibrotic diseases, including those of late onset, often yield unanticipated insights into disease pathogenesis. This Review focuses on pathways underlying lung fibrosis that are generalizable to other organs. Herein, we discuss genetic variants subdivided into those that shorten telomeres, activate the DNA damage response, change resident protein expression or function, or affect organelle activity. Genetic studies provide a window into the downstream cascade of maladaptive responses and pathways that lead to tissue fibrosis. In addition, these studies reveal interactions between genetic variants, environmental factors, and age that influence the phenotypic spectrum of disease. The discovery of forces counterbalancing inherited risk alleles identifies potential therapeutic targets, thus providing hope for future prevention or reversal of fibrosis.


Time to re-evaluate the risks and benefits of valproate and a call for action.
Valproate is widely used in psychiatry and neurology, including off-label use. Here we consider its potential benefits and risks, particularly for women of childbearing potential, and the evidence that clinical guidelines are adhered to. Finally, we consider the implications for clinical practice and research into its efficacy in off-label indications.


Anticonvulsive effects of kappa-opioid receptor modulation in an animal model of ethanol withdrawal.
Although the neurochemical mechanisms contributing to alcohol withdrawal seizures are poorly understood, withdrawal seizures probably reflect neuronal hyperexcitability resulting from adaptation to chronic alcohol. Altered kappa-Opioid receptor (KOP-R) signaling has been observed in multiple seizure types; however, a role for this system in ethanol withdrawal seizures has not been systematically characterized. We hypothesized that pharmacological manipulations of the KOP-R would alter withdrawal in mice selectively bred for differences in ethanol withdrawal severity. Withdrawal Seizure-Prone (WSP) and Withdrawal Seizure-Resistant (WSR) mice were made physically dependent using chronic ethanol vapor inhalation, and the effects of the KOP-R antagonist nor-binaltorphimine or agonist U-50,488H on withdrawal severity were examined. Pretreatment with nor-binaltorphimine significantly increased handling-induced convulsion (HIC) severity in withdrawing WSR mice, with no observable effects in withdrawing WSP mice. In contrast, U-50,488H significantly decreased HIC severity in WSP mice, with no effects in WSR mice. During extended withdrawal (i.e. hours 12+), a rebound hyperexcitability was observed in WSP mice given agonist. Thus, administration of a KOP-R antagonist increased withdrawal severity in mice normally resistant to withdrawal seizures, while a KOP-R agonist reduced convulsion severity in animals susceptible to withdrawal seizures. These observations are consistent with differences in the KOP-R system observed in these lines at the molecular level, and suggest the KOP-R system may be a promising therapeutic target for management of ethanol withdrawal seizures. Finally, these findings underscore the importance of determining the potential for rebound increases in withdrawal severity during later withdrawal episodes.


Global cardiovascular research output, citations, and collaborations: a time-trend, bibliometric analysis (1999-2008).
Health research is one mechanism to improve population-level health and should generally match the health needs of populations. However, there have been limited data to assess the trends in national-level cardiovascular research output, even as cardiovascular disease [CVD] has become the leading cause of morbidity and mortality worldwide. We performed a time trends analysis of cardiovascular research publications (1999-2008) downloaded from Web of Knowledge using a iteratively-tested cardiovascular bibliometric filter with >90% precision and recall. We evaluated cardiovascular research publications, five-year running actual citation indices [ACIs], and degree of international collaboration measured through the ratio of the fractional count of addresses from one country against all addresses for each publication. Global cardiovascular publication volume increased from 40 661 publications in 1999 to 55 284 publications in 2008, which represents a 36% increase. The proportion of cardiovascular publications from high-income, Organization for Economic Cooperation and Development [OECD] countries declined from 93% to 84% of the total share over the study period. High-income, OECD countries generally had higher fractional counts, which suggest less international collaboration, than lower income countries from 1999-2008. There was an inverse relationship between cardiovascular publications and age-standardized CVD morbidity and mortality rates, but a direct, curvilinear relationship between cardiovascular publications and Human Development Index from 1999-2008. Cardiovascular health research output has increased substantially in the past decade, with a greater share of citations being published from low- and middle-income countries. However, low- and middle-income countries with the higher burdens of cardiovascular disease continue to have lower research output than high-income countries, and thus require targeted research investments to improve cardiovascular health.


Monoclonal antibodies in immunosuppression.
Experimental and clinical studies have been carried out to investigate the application of a number of monoclonal antibodies which recognise human lymphocytes in clinical organ transplantation. Where possible, preliminary studies have been carried out using a primate renal allograft model before proceeding to clinical pilot studies. The Campath-1 antigen (CDW52) appears to be a good target for antilymphocytic therapy; a powerful immunosuppressive effect has been demonstrated using both IgM and IgG2b antibodies which recognise this antigen. An IgG2b antibody which recognises the IL-2 receptor was shown to be immunosuppressive in the primate model but clinical studies have, as yet not demonstrated a significant, beneficial effect in preventing rejection. Patients who receive antilymphocyte monoclonal antibodies require monitoring of the expression of the target antigen, the level of free antibody in serum and the development of an antiglobulin response. The current issues in the use of antilymphocyte monoclonal antibodies in clinical immunosuppression include identification of the optimum target, interaction of the antibody with recipient effector mechanisms and methods to avoid or suppress the antiglobulin response.


When do rotator cuff repairs fail? Serial ultrasound examination after arthroscopic repair of large and massive rotator cuff tears.
Despite advances in arthroscopic repair of rotator cuff tears, recurrent tears after repair of large and massive tears remain a significant clinical problem. The primary objective of this study was to define the timing of structural failure of surgically repaired large and massive rotator cuff tears by serial imaging with ultrasound. The secondary objective of this study was to investigate the association between recurrent tears and clinical outcome after rotator cuff repair. Recurrent tear after arthroscopic repair of large rotator cuff tears is more likely to occur late (>3 months) in the postoperative period and will be associated with inferior clinical outcome scores. Cohort study; Level of evidence, 3. Twenty-two consecutive patients with large (>3 cm) rotator cuff tears underwent arthroscopic repair with a standardized technique. Serial ultrasound examinations were performed at 2 days, 2 weeks, 6 weeks, 3 months, 6 months, 12 months, and 24 months after surgery. Western Ontario Rotator Cuff (WORC) Index scores were also collected at these time points. Nine (41%) of the 22 arthroscopically repaired rotator cuff tears demonstrated recurrent tears. Seven of the 9 retears occurred within 3 months of surgery, and the other 2 occurred between 3 and 6 months. No retears occurred after 6 months. At 24-month follow-up, WORC scores favoring intact rotator cuffs over retears approached statistical significance (mean WORC intact 123.9 vs retear 659.8; P = .07). Recurrent rotator cuff tears are not uncommon after arthroscopic repair of large and massive tears. These recurrent tears appear to occur more frequently in the early postoperative period (within the first 3 months) and are associated with inferior clinical outcomes.


Cranberry Extract Standardized for Proanthocyanidins Alleviates β-Amyloid Peptide Toxicity by Improving Proteostasis Through HSF-1 in Caenorhabditis elegans Model of Alzheimer's Disease.
A growing body of evidence suggests that nutraceuticals with prolongevity properties may delay the onset of Alzheimer's disease (AD). We recently demonstrated that a proanthocyanidins-standardized cranberry extract has properties that prolong life span and promote innate immunity in Caenorhabditis elegans In this article, we report that supplementation of this cranberry extract delayed Aβ toxicity-triggered body paralysis in the C elegans AD model. Genetic analyses indicated that the cranberry-mediated Aβ toxicity alleviation required heat shock transcription factor (HSF)-1 rather than DAF-16 and SKN-1. Moreover, cranberry supplementation increased the transactivity of HSF-1 in an IIS-dependent manner. Further studies found that the cranberry extract relies on HSF-1 to significantly enhance the solubility of proteins in aged worms, implying an improved proteostasis in AD worms. Considering that HSF-1 plays a pivotal role in maintaining proteostasis, our results suggest that cranberry maintains the function of proteostasis through HSF-1, thereby protecting C elegans against Aβ toxicity. Together, our findings elucidated the mechanism whereby cranberry attenuated Aβ toxicity in C elegans and stressed the significance of proteostasis in the prevention of age-related diseases from a practical point of view.


Transfection of non-susceptible cells with Ovis aries recombinant lymphocyte function-associated antigen 1 renders susceptibility to Mannheimia haemolytica leukotoxin.
Mannheimia haemolytica is an important etiological agent of pneumonia in domestic sheep (DS, Ovis aries). Leukotoxin (Lkt) produced by this organism is the principal virulence factor responsible for the acute inflammation and lung injury characteristic of M. haemolytica caused disease. Previously, we have shown that the leukocyte-specific integrins, beta(2) integrins, serve as the receptor for Lkt. Although it is certain that CD18, the beta subunit of beta(2) integrins, mediates Lkt-induced cytolysis of leukocytes, it is not clear whether CD18 of all three beta(2) integrins, LFA-1, Mac-1 and CR4, mediates Lkt-induced cytolysis of DS leukocytes. Since polymorphonuclear leukocytes, which express all three beta(2) integrins, are the leukocyte subset that is most susceptible to Lkt, we hypothesized that all three beta(2) integrins serve as the receptor for Lkt. The objective of this study was to determine whether DS LFA-1 serves as a receptor for M. haemolytica Lkt. We cloned the cDNA for DS CD11a, the alpha subunit of LFA-1, and co-transfected it along with the previously cloned cDNA for DS CD18, into a Lkt-non-suceptible cell line. Transfectants stably expressing DS LFA-1 were bound by Lkt. More importantly, Lkt lysed the DS LFA-1 transfectants in a concentration-dependent manner. Pre-incubation of Lkt with a Lkt-neutralizing monoclonal antibody (MAb), or pre-incubation of transfectants with MAbs specific for DS CD11a or CD18, inhibited Lkt-induced cytolysis of the transfectants. Exposure of LFA-1 transfectants to low concentrations of Lkt resulted in elevation of intracellular [Ca(2+)](i). Taken together, these results indicate that DS LFA-1 serves as a receptor for M. haemolytica Lkt.


Continuum model of tendon pathology - where are we now?
Chronic tendon pathology is a common and often disabling condition, the causes of which remain poorly understood. The continuum model of tendon pathology was proposed to provide a model for the staging of tendon pathology and to assist clinicians in managing this often complex condition (Br. J. Sports Med., 43, 2009, 409). The model presents clinical, histological and imaging evidence for the progression of tendon pathology as a three-stage continuum: reactive tendinopathy, tendon disrepair and degenerative tendinopathy. It also provides clinical information to assist in identifying the stage of pathology, in addition to proposed treatment approaches for each stage. The usefulness of such a model is determined by its ability to incorporate and inform new and emerging research. This review examines the degree to which recent research supports or refutes the continuum model and proposes future directions for clinical and research application of the model.


Structure of zirconocene complexes relevant for olefin catalysis: infrared fingerprint of the Zr(C(5)H(5))(2)(OH)(CH(3)CN)(+) cation in the gas phase.
Cationic zirconocene complexes are active species in Ziegler-Natta catalysis for olefin polymerization. Their structure and metal-ligand bond strength strongly influence their activity. In the present work, the infrared multiphoton dissociation (IRMPD) spectrum of mass selected Zr(C(5)H(5))(2)(OH)(CH(3)CN)(+) cations was obtained in the 300-1500 cm(-1) fingerprint range by coupling a Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometer equipped with an electrospray ionization (ESI) source and the infrared free electron laser (IR-FEL) at the Centre Laser Infrarouge d'Orsay (CLIO). The experimental efforts are complemented by quantum chemical calculations at the MP2 and B3LYP levels using the 6-311G* basis set. Vibrational assignments of transitions observed in the IRMPD spectra to modes of the Zr-O-H, C(5)H(5), and CH(3)CN moieties are based on comparison to calculated linear absorption spectra. Both the experimental data and the calculations provide unprecedented information about structure, metal-ligand bonding, charge distribution, and binding energy of the complex.


Childhood obesity: a systems medicine approach.
Childhood obesity and its sequelae are a major public health problem in both the USA and globally. This review will focus on a systems medicine approach to obesity. Systems medicine is an integrative approach utilizing the vast amount of data garnered from "omics" technology and integrating these data with conventional pathophysiology as well as diverse environmental factors such as diet, exercise, community dynamics and the intestinal microbiome. Omics technology includes genomics, epigenomics, metagenomics, metabolomics and proteomics. In addition to unraveling etiology, the goals of a systems medicine approach are to provide actionable and evidenced-based clinical approaches. In the case of childhood obesity, an additional goal is characterizing measureable risk factors/biomarkers for obesity at the earliest possible age and devising age-appropriate optimal intervention strategies. It is also important to establish the age at which interventions could be critical. As discussed below, it is possible that some of the pathophysiological and epigenetic changes resulting from childhood obesity could become more irreversible the longer the obesity remains untreated.


Long noncoding RNA: a new player of heart failure?
One the most important discoveries of the post-genomic era is that a large fraction of the genome transcribes a heterogeneous population of noncoding RNAs (ncRNA). ncRNAs shorter than 200 nucleotides are usually identified as short/small ncRNAs--examples include PIWI-interacting RNAs, small interfering RNAs, and microRNAs (miRNAs)--whereas those longer than 200 nucleotides are classified as long ncRNAs (lncRNAs). These molecules are emerging as important regulators of cellular process, such as development, differentiation, and metabolism. Not surprisingly, ncRNAs are involved also in human diseases, such as cancer and metabolic and neuronal disorders. Although the role of miRNAs is being largely investigated in cardiovascular biology, little is known about other classes of ncRNA in this field. However, recent reports have started to reveal the importance of lncRNA in heart development and suggest also an involvement in heart failure. Here, we will discuss these reports and the therapeutic potential of lncRNA for heart failure.


Microbiological assessment of the implant-abutment interface in different connections: cross-sectional study after 5 years of functional loading.
To evaluate the bacterial microflora present inside the implant connection and in the peri-implant sulcus fluid of healthy implants, and to analyze the relationships between these harboring sites for four different implant systems after at least 5 years of functional loading. A cross-sectional study was performed involving 40 patients treated with metal-ceramic cemented bridges supported by at least two healthy implants functionally loaded for 5 years. Four different implant-abutment connections were studied: external hexagon (control group), double internal hexagon (test group 1), internal hexagon with external collar (test group 2), and conical connection (test group 3). Samples for microbiological analysis were obtained from three types of sites: peri-implant sulci, connections' inside and abutments surface and, as control, gingival sulci of neighboring teeth. Quantitative real-time PCR was carried out for Total Bacterial Count and for 10 microorganisms: Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Campylobacter rectus, Eikenella corrodens, and Candida albicans. The response variables were percentage of positive sites and absolute bacterial load. The relations of the response variables with the type of connection and of sampling site were assessed using generalized estimating equations. Regarding the analysis of positivity to bacteria in the peri-implant sulcus no significant differences were observed. Analyzing the connection's inside, none of the connection designs had the capacity to prevent microbiological leakage through the implant/abutment microgap. Test group 3 presented the lowest mean values for red complex bacteria and control group the highest, although differences were non-significant. Statistical significance was only reached for Treponema denticola in the bacterial load analysis inside the connection. Test groups 1 and 2 yielded lower values for orange complex bacteria but only for Peptostreptococos micros the differences resulted significant. Test groups 2 and 3 had significantly lower total bacterial counts in the peri-implant sulcus and inside the connection. Outcomes suggested that all the analyzed connections resulted contaminated after 5 years of functional loading. However, the connection design might influence bacterial activity levels qualitatively and quantitatively, especially inside the implant connection.


Digital fluoroscopy as a valuable adjunct to open vascular operations.
The increasing availability of and vascular surgeons' familiarity with digital cine-fluoroscopy in the operating room has been facilitated by the advent and growing popularity of endovascular aortoiliac aneurysm repair and other endovascular techniques that are being incorporated into vascular surgical practice. Digital cine-fluoroscopy can also be used as a valuable adjunct to standard open vascular procedures in several ways including: performance of completion angiography, fluoroscopically-assisted thromboembolectomy, intraoperative planning angiography, fluoroscopically-guided pressure gradient measurements, achieving vascular control of proximal arteries, intraoperative thrombolysis of compromised outflow tracts, and angioplasty and stenting of lesions detected intraoperatively. These techniques can improve the outcome of standard vascular procedures by permitting the identification of inflow, outflow, conduit, and anastomotic defects intraoperatively and guiding their repair. Additionally, in many cases they can reduce the amount of exposure required, reduce intraoperative blood loss, and minimize trauma to vessels during thrombectomy. Fluoroscopic guidance can facilitate and improve these and other aspects of standard open vascular procedures. Conversely, the ability to perform open interventions can facilitate the performance of many endovascular interventions. It is becoming increasingly important to be facile with both open and E fluoroscopically guided techniques in order to fully treat the spectrum of vascular disease in an optimum fashion.


Finding paradise: cues directing the migration of the waterfall climbing Hawaiian gobioid Sicyopterus stimpsoni.
A series of waterfall-climbing trials were conducted to identify cues that direct the climbing of juvenile Sicyopterus stimpsoni. In the first experiment, whether climbing juveniles preferentially ascend water sources with conspecifics or whether the presence of just stream water is sufficient to attract fish to ascend a climbing path were assessed. In the second experiment, whether climbing juveniles create a trail of mucus that facilitates the ability of conspecifics to follow their lead was determined. The results indicate that juvenile S. stimpsoni are less likely to climb in waters devoid of organic cues but are strongly attracted to stream water with or without the odour of conspecifics. Once climbing, performance did not differ for juveniles climbing in differing water choices, suggesting an all-or-nothing commitment once climbing commences. Climbing S. stimpsoni did produce a mucous trail while climbing that was associated with a mucous gland that dramatically increases in size just prior to juveniles gaining the ability to climb. The trail was not followed closely by subsequent juveniles traversing the same channel, however, suggesting only weak trail-following in waterfall climbing S. stimpsoni. Previous genetic studies suggest that juvenile S. stimpsoni do not home to natal streams in the face of strong near-shore oceanic currents. Instead, these fish appear primarily to rely on cues that suggest the presence of organic growth in streams, a factor that may indicate suitable habitat in an ever-changing stream environment but which may also be vulnerable to interference through human activity.


Current treatment for liver metastases from colorectal cancer.
The liver is the commonest site of distant metastasis of colorectal cancer and nearly half of the patients with colorectal cancer ultimately develop liver involved during the course of their diseases. Surgery is the only therapy that offers the possibility of cure for patients with hepatic metastatic diseases. Five-year survival rates after resection of all detectable liver metastases can be up to 40 %. Unfortunately, only 25 % of patients with colorectal liver metastases are candidates for liver resection, while the others are not amenable to surgical resection. Regional therapies such as radiofrequency ablation and cryotherapy may be offered to patients with isolated unresectable metastases but no extrahepatic diseases. Hepatic artery catheter chemotherapy and chemoembolization and portal vein embolization are often used for the patients with extensive liver metastases but without extrahepatic diseases, which are not suitable for regional ablation. For the patients with metastatic colorectal cancer beyond the liver, systemic chemotherapy is a more appropriate choice. Immunotherapy is also a good option when other therapies are used in combination to enhance the efficacy. Selective internal radiation therapy is a new radiation method which can be used in patients given other routine therapies without effects.


Living with high putrescine: expression of ornithine and arginine biosynthetic pathway genes in high and low putrescine producing poplar cells.
Arginine (Arg) and ornithine (Orn), both derived from glutamate (Glu), are the primary substrates for polyamine (PA) biosynthesis, and also play important roles as substrates and intermediates of overall N metabolism in plants. Their cellular homeostasis is subject to multiple levels of regulation. Using reverse transcription quantitative PCR (RT-qPCR), we studied changes in the expression of all genes of the Orn/Arg biosynthetic pathway in response to up-regulation [via transgenic expression of mouse Orn decarboxylase (mODC)] of PA biosynthesis in poplar (Populus nigra × maximowiczii) cells grown in culture. Cloning and sequencing of poplar genes involved in the Orn/Arg biosynthetic pathway showed that they have high homology with similar genes in other plants. The expression of the genes of Orn, Arg and PA biosynthetic pathway fell into two hierarchical clusters; expression of one did not change in response to high putrescine, while members of the other cluster showed a shift in expression pattern during the 7-day culture cycle. Gene expression of branch point enzymes (N-acetyl-Glu synthase, Orn aminotransferase, Arg decarboxylase, and spermidine synthase) in the sub-pathways, constituted a separate cluster from those involved in intermediary reactions of the pathway (N-acetyl-Glu kinase, N-acetyl-Glu-5-P reductase, N-acetyl-Orn aminotransferase, N (2)-acetylOrn:N-acetyl-Glu acetyltransferase, N (2)-acetyl-Orn deacetylase, Orn transcarbamylase, argininosuccinate synthase, carbamoylphosphate synthetase, argininosuccinate lyase, S-adenosylmethionine decarboxylase, spermine synthase). We postulate that expression of all genes of the Glu-Orn-Arg pathway is constitutively coordinated and is not influenced by the increase in flux rate through this pathway in response to increased utilization of Orn by mODC; thus the pathway involves mostly biochemical regulation rather than changes in gene expression. We further suggest that Orn itself plays a major role in the regulation of this pathway.