The significance of circulating and cell-bound antibodies in experimental allergic encephalomyelitis.
Conjugates of horseradish peroxidase with myelin basic protein (BP) of guinea pig or Lewis rat were used to identify antibody-containing cells in draining lymph nodes during experimental allergic encephalomyelitis (EAE). Peroxidase activity was revealed for light and electron microscopic preparations with the diaminobenzidine reaction of Graham and Karnovsky. Basic proteins (BP) were also iodinated with (125)I for determination of circulating antibody against BP by radio-immunoassay of (125)I BP using coprecipitation with antirat IgG or with antirat serum proteins. Encephalitogenicity was lost after conjugation of guinea pig BP or Lewis rat BP with peroxidase, whereas iodination did not affect the encephalitogenicity of guinea pig or Lewis rat BPs. EAE was induced in Lewis rats with guinea pig or Lewis rat spinal cord BPs in complete Freund's adjuvant. Draining lymph nodes were studied by light and electron microscopy during the course of the immune reaction, and cells with specific antibody against BP were identified with the use of BP-horseradish peroxidase conjugates. Lymph node sections from animals immunized with high antigen doses (500 mug) showed numerous plasma cells with intracellular antibody against BP in medullary cords 10 days after immunization and 4 days prior to histologic appearance of EAE. Numbers of positive cells correlated with levels of circulating antibody against BP. Immunization with a low antigen dose (5 mug) resulted in EAE, few or no antibody-containing cells, and significantly lower levels of circulating antibody. Brown Norwegian rats, a strain resistant to EAE, immunized with 500 mug of BP had positive cells in draining lymph nodes and high levels of circulating antibody against BP in the absence of histologic evidence of EAE. Lewis rats injected with Lewis rat small BP failed to develop EAE. Nevertheless, these animals showed levels of circulating antibody and antibody-containing cells similar to those of animals which developed EAE after injection of the mixture of Lewis rat large and small BP. It is concluded that although the BP-peroxidase labeling method reveals cells with specific anti-BP antibody, these cells are probably unrelated to EAE. The lack of correlation between EAE induced by low antigen doses and levels of circulating anti-BP antibody (determined with the use of highly encephalitogenic (125)I-BP) suggests that effector cells can be stimulated at low antigen doses, but higher antigen doses are required to induce the production of levels of circulating antibody detectable by the method of immune coprecipitation.


Characterization of Klebsiella pneumoniae type 1 fimbriae by detection of phase variation during colonization and infection and impact on virulence.
Klebsiella pneumoniae is recognized as an important gram-negative opportunistic pathogen. The ability of bacteria to adhere to host structures is considered essential for the development of infections; however, few studies have examined the influence of adhesion factors on K. pneumoniae virulence. In this study, we cloned and characterized the type 1 fimbria gene cluster of a clinical K. pneumoniae isolate. Although this cluster was not identical to the Escherichia coli type 1 fimbria gene cluster, an overall high degree of structural resemblance was demonstrated. Unique to the K. pneumoniae fim gene cluster is the fimK gene, whose product contains an EAL domain, suggesting that it has a role in regulation of fimbrial expression. Like expression of type 1 fimbriae in E. coli, expression of type 1 fimbriae in K. pneumoniae was found to be phase variable, and an invertible DNA element (fim switch) was characterized. An isogenic type 1 fimbria mutant was constructed and used to evaluate the influence of type 1 fimbriae in different infection models. Type 1 fimbriae did not influence the ability of K. pneumoniae to colonize the intestine or infect the lungs, but they were determined to be a significant virulence factor in K. pneumoniae urinary tract infection. By use of a PCR-based assay, the orientation of the fim switch during colonization and infection was investigated and was found to be all "off" in the intestine and lungs but all "on" in the urinary tract. Our results suggest that during colonization and infection, there is pronounced selective pressure in different host environments for selection of either the type 1 fimbriated or nonfimbriated phenotype of K. pneumoniae.


A systematic and transparent approach for assessing the methodological quality of intervention effectiveness research: the Study Design and Implementation Assessment Device (Study DIAD).
Assessments of studies meant to evaluate the effectiveness of interventions, programs, and policies can serve an important role in the interpretation of research results. However, evidence suggests that available quality assessment tools have poor measurement characteristics and can lead to opposing conclusions when applied to the same body of studies. These tools tend to (a) be insufficiently operational, (b) rely on arbitrary post-hoc decision rules, and (c) result in a single number to represent a multidimensional construct. In response to these limitations, a multilevel and hierarchical instrument was developed in consultation with a wide range of methodological and statistical experts. The instrument focuses on the operational details of studies and results in a profile of scores instead of a single score to represent study quality. A pilot test suggested that satisfactory between-judge agreement can be obtained using well-trained raters working in naturalistic conditions. Limitations of the instrument are discussed, but these are inherent in making decisions about study quality given incomplete reporting and in the absence of strong, contextually based information about the effects of design flaws on study outcomes.


Empyema: analysis of treatment techniques.
Empyema is associated with a high mortality. To study the factors responsible for the failure of treatment, the authors reviewed 90 cases of nontuberculous thoracic empyema seen at the University of Western Ontario in London, between 1970 and 1980. The most common causes of empyema were bronchopulmonary infections (63%), complications of pulmonary surgery (14%) and secondary infections of hydrothoraces or hemothoraces (13%). In 51 patients (57%) the condition was acquired in hospital or was related to previous medical therapy. Nine cases were recognized only at autopsy. The treatment of empyema was assessed in 81 patients, who received an average of 2.2 antibiotics during the course of treatment. Five patients received antibiotics as the only therapy; one died. Seventy-two patients were treated by surgical drainage; 35 (49%) were cured and 18 (25%) subsequently died. Twelve of 18 decortication procedures were successful including 4 performed as a primary procedure and 8 as a secondary procedure. Five of seven patients who underwent thoracoplasty were cured. The overall mortality in the series was 23%. Prevention and early recognition of empyema may reduce the mortality. Patients who do not improve promptly with surgical drainage may benefit from early decortication.


[Obesity: concept, classification and diagnosis].
Obesity is a chronic disease that is characterised by an increase of fat mass and as a result by an increase in weight. There is therefore an increase in the energy reserves of the organism in the form of fat. The term chronic is applied due to its forming part of the group of diseases that we are unable to cure with the therapeutic arsenal that is now available. From an anthropometric point of view, which is habitually used in the clinic, a person is considered to be obese with a Body Mass Index equal to or higher than 30 kg/m2. To be able to evaluate obesity account must be taken not only of the anthropometric aspects but also of the possible genetic factors; the causes of the disease must be studied and the possible existence of associated complications and diseases must be checked. Treatment must always be personalised and adapted to the characteristics and comorbidities presented by the patient. The dominant criteria favourable to therapeutic intervention in obesity are especially based on the demonstration that with a moderate loss of body weight (5-10 %) a notable improvement can be obtained in the comorbidity associated with obesity and in the quality of life of the obese patient.


Defined bacterial populations in the rumens of gnotobiotic lambs.
Five gnotobiotic lambs were fed on sterile diets until they were killed at 13 to 21 weeks of age. They were dosed orally with different combinations of 11 species of rumen bacteria. The biochemical reactions of each of the bacteria inoculated had been determined in pure culture in vitro, and they were chosen to perform the main reactions known to be associated with digestion in the normal mature rumen. Two of the bacteria could not be reisolated, but the remainder had established readily in the rumen, forming stable, mixed, defined populations. The total numbers of bacteria in the rumen, and the viable counts of most of the individual species were comparable to those of normal sheep. The concentration of volatile fatty acids was lower, however, and in four of the lambs there was a higher proportion of butyric acid and a lower proportion of propionic acid than in normal sheep. Cellulolytic, ureolytic, and methanogenic activities appeared to be taking place and lactate-utilizing bacteria appeared to reverse the accumulation of lactate which resulted from the activity of lactate-producing bacteria. Some of the bacteria also established at high levels in the caecum.


Method of induction could affect emotional outcomes: comment on markey, chin, vanepps, and loewenstein (2014 ).
Markey, Chin, Vanepps, and Loewenstein (2014) demonstrated six methods for the induction of boredom. However, a clear and testable definition of boredom should be established prior to experimental manipulation of the construct. Defining boredom from a functional emotion perspective is one approach that affords a definition separable from the outcomes associated with boredom and insight into which manipulations may effectively target the construct.


Interactions of Plasmodium berghei sporozoites and murine Kupffer cells in vitro.
Malaria sporozoites must leave the bloodstream and cross a layer of sinusoidal lining cells in order to infect hepatocytes and undergo exoerythrocytic schizogony. To determine whether Kupffer cells (KC) derived from this layer interact with sporozoites, murine KC were isolated from perfused livers of BALB/cJ mice and incubated in vitro with Plasmodium berghei sporozoites. Isolated KC had characteristic macrophage surface Ag and were phagocytic, ingesting both latex particles and Leishmania major amastigotes. In the absence of immune serum, sporozoites associated with fewer than 10% of these KC. By 30 min, 10% of the cell-associated sporozoites were completely ingested, 30% were in the process of being ingested, and 60% were attached to the surface of the cells. Opsonization of sporozoites with monoclonal or polyclonal antibodies directed against P. berghei circumsporozoite protein markedly enhanced sporozoite association with KC. Up to 40% of cells exposed to opsonized sporozoites had parasites inside or attached to their surfaces. Sporozoites attached to or ingested by KC were uniformly destroyed within 240 min in all cultures; there was no evidence of conversion of sporozoites to the exoerythrocytic stage within KC by light microscopy, and there was no evidence of residual sporozoites, either inside or outside of cells, by either light or electron microscopy. These data suggest that under nonimmune conditions, KC play a minor role in resistance to infection by malaria sporozoites. However, when sporozoites are opsonized by circumsporozoite antibodies, phagocytosis by KC may be an important immune mechanism that prevents parasitization of hepatocytes.


Giant cell tumour of the hamate. Case report.
We report the case of a 31-year-old woman who had had the distal end of the ulna resected for a giant cell tumour five years before presenting with a further one in the hamate. The medullary cavity of the hamate was curetted and cancellous bone grafted from the distal radius. Her symptoms resolved, and 26 months postoperatively the cortex of the hamate was fully restored.


Effects of secretagogues on [32P]phosphatidylinositol 4,5-bisphosphate metabolism in the exocrine pancreas.
Experiments were carried out to assess the effects of secretagogues on the polyphosphoinositides phosphatidylinositol 4-phosphate (PtdIns4P) and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] on preparations of exocrine pancreas in vitro. Carbachol and caerulein provoked a rapid (less than 1 min) breakdown of 15-20% of [32P]PtdIns(4,5)P2 in isolated pancreatic acini, but did not affect [32P]PtdIns4P. In contrast, the Ca2+ ionophore ionomycin had no immediate effect on the levels of either inositide but caused a parallel fall in both lipids after 5-10 min. A similar decrease in [32P]PtdIns(4,5)P2 due to carbachol was obtained with isolated acini and isolated cells, despite the fact that the secretory response of isolated cells was considerably less than that of isolated acini. Loss of [32P]PtdIns(4,5)P2 elicited by carbachol or caerulein was unaffected either by the addition of EGTA in excess of extracellular Ca2+ or when a protocol was employed that eliminated caerulein-induced intracellular Ca2+-release. These results suggest that agonist-induced PtdIns(4,5)P2 breakdown in the exocrine pancreas may be an early step in the stimulus-response coupling pathway and also suggest that this breakdown is not dependent on Ca2+-mobilization.


Interplay Between Angiotensin II Type 1 Receptor and Thrombin Receptor Revealed by Bioluminescence Resonance Energy Transfer Assay.
The key hormone of the renin-angiotensin system (RAS), angiotensin II (AngII), and thrombin are known to play major roles in the vascular system and its related disorders. Previous studies reported connections between AngII and thrombin in both physiological and pathophysiological models. However, the molecular mechanisms controlling such interplay at the level of their receptors belonging to the family of G protein-coupled receptors (GPCRs) are not fully understood. In this study, we investigated the functional interaction between the AngII type 1 receptor (AT1R) and the thrombin receptor [or protease-activated receptor 1 (PAR1)] in human embryonic kidney 293 (HEK293) cells. For this, we used various bioluminescence resonance energy transfer (BRET) proximity-based assays to profile the coupling to the heterotrimeric Gαq protein, β-arrestin recruitment, and receptor internalization and trafficking in intact cells. The overall dose-response and real-time kinetic BRET data demonstrated the specific molecular proximity between AT1R and PAR1 resulting in their functional interaction. This was characterized by thrombin inducing BRET increase within AT1R/Gαq and AT1R/β-arrestin pairs and synergistic effects observed upon the concomitant activation of both receptors suggesting a positive allosteric interaction. The BRET data corroborated with the data on the downstream Gαq/inositol phosphate pathway. Moreover, the selective pharmacological blockade of the receptors revealed the implication of both AT1R and PAR1 protomers in such a synergistic interaction and the possible transactivation of AT1R by PAR1. Interestingly, the positive action of PAR1 on AT1R activation was contrasted with its apparent inhibition of AT1R internalization and its endosomal trafficking. Finally, BRET saturation and co-immunoprecipitation assays supported the physical AT1-PAR1 interaction in HEK293 cells. Our study reveals for the first time the functional interaction between AT1R and PAR1 in vitro characterized by a transactivation and positive allosteric modulation of AT1R and inhibition of its desensitization and internalization. This finding may constitute the molecular basis of the well-known interplay between RAS and thrombin. Thus, our data should lead to revising some findings on the implication of RAS and thrombin in vascular physiology and pathophysiology revealing the importance to consider the functional and pharmacological interaction between AT1R and thrombin receptors.


Use of lambda gt11 to isolate genes for two pseudorabies virus glycoproteins with homology to herpes simplex virus and varicella-zoster virus glycoproteins.
A library of pseudorabies virus (PRV) DNA fragments was constructed in the expression cloning vector lambda gt11. The library was screened with antisera which reacted with mixtures of PRV proteins to isolate recombinant bacteriophages expressing PRV proteins. By the nature of the lambda gt11 vector, the cloned proteins were expressed in Escherichia coli as beta-galactosidase fusion proteins. The fusion proteins from 35 of these phages were purified and injected into mice to raise antisera. The antisera were screened by several different assays, including immunoprecipitation of [14C]glucosamine-labeled PRV proteins. This method identified phages expressing three different PRV glycoproteins: the secreted glycoprotein, gX; gI; and a glycoprotein that had not been previously identified, which we designate gp63. The gp63 and gI genes map adjacent to each other in the small unique region of the PRV genome. The DNA sequence was determined for the region of the genome encoding gp63 and gI. It was found that gp63 has a region of homology with a herpes simplex virus type 1 (HSV-1) protein, encoded by US7, and also with varicella-zoster virus (VZV) gpIV. The gI protein sequence has a region of homology with HSV-1 gE and VZV gpI. It is concluded that PRV, HSV, and VZV all have a cluster of homologous glycoprotein genes in the small unique components of their genomes and that the organization of these genes is conserved.


2020 Roadmap on Carbon Materials for Energy Storage and Conversion.
Carbon is a simple, stable and popular element with many allotropes. The carbon family members include carbon dots, carbon nanotubes, carbon fibers, graphene, graphite, graphdiyne and hard carbon, etc. They can be divided into different dimensions, and their structures can be open and porous. Moreover, it is very interesting to dope them with other elements (metal or non-metal) or hybridize them with other materials to form composites. The elemental and structural characteristics offer us to explore their applications in energy, environment, bioscience, medicine, electronics and others. Among them, energy storage and conversion are extremely attractive, as advances in this area may improve our life quality and environment. Some energy devices will be included herein, such as lithium-ion batteries, lithium sulfur batteries, sodium-ion batteries, potassium-ion batteries, dual ion batteries, electrochemical capacitors, and others. Additionally, carbon-based electrocatalysts are also studied in hydrogen evolution reaction and carbon dioxide reduction reaction. However, there are still many challenges in the design and preparation of electrode and electrocatalytic materials. The research related to carbon materials for energy storage and conversion is extremely active, and this has motivated us to contribute with a roadmap on 'Carbon Materials in Energy Storage and Conversion'.


Genetic pharmacology: progresses in siRNA delivery and therapeutic applications.
In the RNA interference process, the catalytic degradation of an endogenous mRNA results from the Watson-Crick complementary recognition by either a small silencing synthetic double-stranded ribonucleotide (siRNA) or by a small hairpin RNA (shRNA) produced in the cell by transcription from a DNA template. This interference process ideally results in an exquisitely specific mRNA suppression. The present review is dedicated to siRNAs. It describes the mechanism of RNA silencing and the main siRNA delivery techniques, with a focus on siRNA self-complexing to cationic lipids to form nanoparticles, which are called lipoplexes. The addition to lipoplexes of an anionic polymer leads to the ternary formulation APIRL (Anionic-Polymer-Interfering-RNA-Lipoplexes) with increased in vivo stability and biological efficacy. In terms of clinical development, the review focuses on therapeutic applications by intravenous delivery to the liver and inflammatory joints, and to localized siRNA delivery to the ocular sphere.


[Effect of zhuchun pill on immunity and endocrine function of elderly with kidney-yang deficiency].
After randomly dividing 45 elderly males with Kidney-Yang Deficiency into Zhuchun Pill (ZCP) group and placebo (starch capsule) group by single blind method, and treating them for 3 months, we found that ZCP could greatly improve the main symptoms of Kidney-Yang Deficiency, P < 0.01 by comparison with control group: the 3H-TdR lymphocyte transformation (3H-TdR LCT) rate, level of serum C3, IgA, IgD, IgG & IgM went up (P < 0.01); contents of plasma adrenocortical hormone (ACTH), testosterone (T) and cyclic adenosine monophosphate (cAMP) increased (P < 0.01-0.001): the level of superoxide dismutase (SOD) in serum also increased while lipid peroxide (LPO) in plasma decreased (P < 0.01). But for control group, there were no remarkable changes after treatment, so it revealed that the improvement of Kidney-Yang Deficiency by ZCP is realized through regulating the organism's immunity and endocrine function as well as through scavenging the free radical agents.


Population-based study of medulloblastoma: outcomes in Alberta from 1975 to 1996.
The purpose of this study was to determine incidence, survival rate, and prognostic factors as well as the frequency of Collins' Law Violators (CLVs) in an unselected population of medulloblastoma patients. Collins' Law dictates that 'cure' of a child with a tumor occurs after a period that includes the child's age at diagnosis plus 9 months. Using the Alberta Cancer Registry a population-based review identified 49 patients with medulloblastoma (19 adults, 30 children) diagnosed from 1975-96. Pathology was reviewed in all cases. All patients had surgical resection, followed by radiotherapy in 47 patients and chemotherapy in 17. The overall 5-year survival was 50%. There was a trend for the extent of resection to be associated with a longer survival (Long rank test, p < 0.06) but this was not significant. Tumor recurrence occurred a median of 22.4 months (range, 6.4-192.3) after diagnosis and median survival after recurrence was 9.3 months (range, 0.4-64.9). The survival curve did not appear to plateau but was affected by tumor-related deaths in 3 (21.4%) of the 21 long-term survivors diagnosed in childhood. These three patients had recurrences a mean of 11.7 years after diagnosis and are designated as CLVs. The survival rate in an unselected population of patients with medulloblastoma is poor. Aggressive resection of the tumors prolongs survival. The Collins' Law Violators were relatively common and we suggest this concept be abandoned in medulloblastoma.


ME-3407, a new antiulcer agent, inhibits acid secretion by interfering with redistribution of H(+)-K(+)-ATPase.
ME-3407 is a newly developed antiulcer drug that markedly promoted the healing of acetic acid-induced chronic ulcers in rats presumably due to potent inhibition of acid secretion. ME-3407 and its metabolites, the sulfoxide of which was preserved, produced dosedependent inhibition of aminopyrine accumulation by rabbit gastric glands stimulated by any agonist, suggesting that the site of their action was downstream from the production of second messengers. Although one of the metabolites, EF-4025, showed some inhibitory effects on functional activities of H(+)-K(+)-ATPase, ME-3407 itself was not a proton pump inhibitor. ME-3407, but not omeprazole, inhibited the stimulation-associated redistribution of H(+)-K(+)-ATPase from microsomes into the apical membranes in addition to delocalizing ezrin, a putative F-actin-membrane linker, from apical plasma membrane. ME-3407 and EF-4025 inhibited myosin light chain kinase (MLCK) and protein kinase A activities. Because another MLCK inhibitor, wortmannin, showed the same properties as ME-3407, i.e., inhibition of aminopyrine accumulation, inhibition of stimulation-associated redistribution of H(+)-K(+)-ATPase, and abnormal distribution of ezrin, we hypothesize that MLCK is one of the potential targets for the drug. We conclude that ME-3407 is a promising drug for treating peptic ulcers, as well as a useful tool for studying mechanisms of parietal cell activation, especially related to the recruitment and recycling of the proton pump.


Trainee Therapists' Perceptions of a Blended Intervention to Promote Resilience after a Natural Disaster: A Qualitative Case Study.
Natural disasters happen in an increased frequency, and telemental health interventions could offer easily accessible help to reduce mental health symptoms experienced by survivors. However, there are very few programs offered to natural disaster survivors, and no research exists on therapists' experiences with providing blended interventions for natural disaster survivors. Our qualitative case study aims to describe psychologists' experiences with an online, therapist-assisted blended intervention for survivors of the Fort McMurray wildfires in Alberta, Canada. The RESILIENT intervention was developed in the frames of a randomized controlled trial to promote resilience after the Fort McMurray wildfires by providing survivors free access to a 12-module, therapist-assisted intervention, aiming to improve post-traumatic stress, insomnia, and depression symptoms. A focus group design was used to collect data from the therapists, and emerging common themes were identified by thematic analysis. Therapists felt they could build strong alliances and communicate emotions and empathy effectively, although the lack of nonverbal cues posed some challenges. The intervention, according to participating therapists, was less suitable for participants in high-stress situations and in case of discrepancy between client expectations and the intervention content. Moreover, the therapists perceived specific interventions as easy-to-use or as more challenging based on their complexity and on the therapist support needed for executing them. Client engagement in the program emerged as an underlying theme that had fundamental impact on alliance, communication, and ultimately, treatment efficiency. Therapist training and supervision was perceived as crucial for the success of the program delivery. Our findings provided several implications for the optimalization of blended interventions for natural disaster survivors from our therapists' perspective.


Appearance of claudin-5+ leukocyte subtypes in the blood and CNS during progression of EAE.
Tight junctions (TJs) are membrane specializations characteristic of barrier-forming membranes, which function to seal the aqueous pathway between endothelial cells or epithelial cells and, thereby, obstruct intercellular solute and cellular movement. However, previous work from our laboratory found that claudin-5 (CLN-5), a TJ protein prominent at the blood-brain barrier (BBB), was also detected, ectopically, on leukocytes (CLN-5+) in the blood and central nervous system (CNS) of mice with experimental autoimmune encephalomyelitis (EAE), a neuroinflammatory, demyelinating disease that is a model for multiple sclerosis. CLN-5 was further shown to be transferred from endothelial cells to circulating leukocytes during disease, prompting consideration this action is coupled to leukocyte transendothelial migration (TEM) into the CNS by fostering transient interactions between corresponding leukocyte and endothelial junctional proteins at the BBB. To begin clarifying the significance of CLN-5+ leukocytes, flow cytometry was used to determine their appearance in the blood and CNS during EAE. Flow cytometric analysis revealed CLN-5+ populations among CD4 and CD8 T cells, B cells, monocytes and neutrophils, and these appeared with varying kinetics and to different extents in both blood and CNS. CLN-5 levels on circulating T cells further correlated highly with activation state. And, the percentage of CLN-5+ cells among each of the subtypes analyzed was considerably higher in CNS tissue than in blood, consistent with the interpretation that CLN-5+ leukocytes gain preferred access to the CNS. Several leukocyte subtypes variably acquire CLN-5 in blood before they enter the CNS, an event that may represent a novel mechanism to guide leukocytes to sites for paracellular diapedesis across the BBB.


Markovian timed Petri nets for performance analysis of semiconductor manufacturing systems.
A subclass of generalized stochastic Petri nets (GSPNs) with priorities, called Markovian timed Petri nets, are proposed to model semiconductor manufacturing systems that consider process priorities, routing priorities, resource re-entrance, and nonpreemptive operations. Uniformization technique is used to establish both lower and upper bounds of the performance of interest. These bounds are computable using linear programming. Numerical experiments have been conducted to evaluate the accuracy of the bounds using models adapted from real-world systems. The experiments show that the upper bounds are very close to the simulation results. Thus, performance measures can be accurately estimated using these bounds.


An Ethylene-Protected Achilles' Heel of Etiolated Seedlings for Arthropod Deterrence.
A small family of Kunitz protease inhibitors exists in Arabidopsis thaliana, a member of which (encoded by At1g72290) accomplishes highly specific roles during plant development. Arabidopsis Kunitz-protease inhibitor 1 (Kunitz-PI;1), as we dubbed this protein here, is operative as cysteine PI. Activity measurements revealed that despite the presence of the conserved Kunitz-motif the bacterially expressed Kunitz-PI;1 was unable to inhibit serine proteases such as trypsin and chymotrypsin, but very efficiently inhibited the cysteine protease RESPONSIVE TO DESICCATION 21. Western blotting and cytolocalization studies using mono-specific antibodies recalled Kunitz-PI;1 protein expression in flowers, young siliques and etiolated seedlings. In dark-grown seedlings, maximum Kunitz-PI;1 promoter activity was detected in the apical hook region and apical parts of the hypocotyls. Immunolocalization confirmed Kunitz-PI;1 expression in these organs and tissues. No transmitting tract (NTT) and HECATE 1 (HEC1), two transcription factors previously implicated in the formation of the female reproductive tract in flowers of Arabidopsis, were identified to regulate Kunitz-PI;1 expression in the dark and during greening, with NTT acting negatively and HEC1 acting positively. Laboratory feeding experiments with isopod crustaceans such as Porcellio scaber (woodlouse) and Armadillidium vulgare (pillbug) pinpointed the apical hook as ethylene-protected Achilles' heel of etiolated seedlings. Because exogenous application of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and mechanical stress (wounding) strongly up-regulated HEC1-dependent Kunitz-PI;1 gene expression, our results identify a new circuit controlling herbivore deterrence of etiolated plants in which Kunitz-PI;1 is involved.


Multi-marker metabarcoding of coral skeletons reveals a rich microbiome and diverse evolutionary origins of endolithic algae.
Bacteria, fungi and green algae are common inhabitants of coral skeletons. Their diversity is poorly characterized because they are difficult to identify with microscopy or environmental sequencing, as common metabarcoding markers have low phylogenetic resolution and miss a large portion of the biodiversity. We used a cost-effective protocol and a combination of markers (tufA, 16S rDNA, 18S rDNA and 23S rDNA) to characterize the microbiome of 132 coral skeleton samples. We identified a wide range of prokaryotic and eukaryotic organisms, many never reported in corals before. We additionally investigated the phylogenetic diversity of the green algae-the most abundant eukaryotic member of this community, for which previous literature recognizes only a handful of endolithic species. We found more than 120 taxonomic units (near species level), including six family-level lineages mostly new to science. The results suggest that the existence of lineages with an endolithic lifestyle predates the existence of modern scleractinian corals by ca. 250my, and that this particular niche was independently invaded by over 20 lineages in green algae evolution. These results highlight the potential of the multi-marker approach to assist in species discovery and, when combined with a phylogenetic framework, clarify the evolutionary origins of host-microbiota associations.


Computed tomography fractional flow reserve discrepancies with invasive angiography and physiology - Case series.
Computed tomography with fractional flow reserve (CT-FFR) is increasingly common in assessing coronary artery disease. We report five cases of discrepancies that led to changes in treatment. This report highlights discordant findings between modalities, which should be considered during the diagnostic assessment of chest pain.


Numbers and organization of RNA polymerases, nascent transcripts, and transcription units in HeLa nuclei.
Using HeLa cells, we have developed methods to determine 1) the number of RNA polymerases that are active at any moment, 2) the number of transcription sites, and 3) the number of polymerases associated with one transcription unit. To count engaged polymerases, cells were encapsulated in agarose, permeabilized, treated with ribonuclease, and the now-truncated transcripts extended in [32P]uridine triphosphate; then, the number of growing transcripts was calculated from the total number of nucleotides incorporated and the average increment in length of the transcripts. Approximately 15, 000 transcripts were elongated by polymerase I, and approximately 75,000 were elongated by polymerases II and III. Transcription sites were detected after the cells were grown in bromouridine for <2.5 min, after which the resulting bromo-RNA was labeled with gold particles; electron microscopy showed that most extranucleolar transcripts were concentrated in approximately 2400 sites with diameters of approximately 80 nm. The number of polymerases associated with a transcription unit was counted after templates were spread over a large area; most extranucleolar units were associated with one elongating complex. These results suggest that many templates are attached in a "cloud" of loops around a site; each site, or transcription "factory," would contain approximately 30 active polymerases and associated transcripts.


Custom-Made Poly(urethane) Coatings Improve the Mechanical Properties of Bioactive Glass Scaffolds Designed for Bone Tissue Engineering.
The replication method is a widely used technique to produce bioactive glass (BG) scaffolds mimicking trabecular bone. However, these scaffolds usually exhibit poor mechanical reliability and fast degradation, which can be improved by coating them with a polymer. In this work, we proposed the use of custom-made poly(urethane)s (PURs) as coating materials for 45S5 Bioglass®-based scaffolds. In detail, BG scaffolds were dip-coated with two PURs differing in their soft segment (poly(ε-caprolactone) or poly(ε-caprolactone)/poly(ethylene glycol) 70/30 w/w) (PCL-PUR and PCL/PEG-PUR) or PCL (control). PUR-coated scaffolds exhibited biocompatibility, high porosity (ca. 91%), and improved mechanical properties compared to BG scaffolds (2-3 fold higher compressive strength). Interestingly, in the case of PCL-PUR, compressive strength significantly increased by coating BG scaffolds with an amount of polymer approx. 40% lower compared to PCL/PEG-PUR- and PCL-coated scaffolds. On the other hand, PEG presence within PCL/PEG-PUR resulted in a fast decrease in mechanical reliability in an aqueous environment. PURs represent promising coating materials for BG scaffolds, with the additional pros of being ad-hoc customized in their physico-chemical properties. Moreover, PUR-based coatings exhibited high adherence to the BG surface, probably because of the formation of hydrogen bonds between PUR N-H groups and BG surface functionalities, which were not formed when PCL was used.


Right ventricular outflow tract growth in infants with palliated tetralogy of fallot.
In symptomatic neonates with tetralogy of Fallot with pulmonary stenosis (TOF/PS), limited contemporary data exist regarding the effect of a modified Blalock-Taussig shunt (mBTS) on pulmonary valve (PV) and pulmonary artery (PA) growth and on the incidence of PV preservation at the time of complete repair. We retrospectively studied patients who underwent repair of TOF/PS from 2000 to 2012 at our center. In neonates with TOF/PS palliated with an mBTS, we assessed PV annulus, main PA (MPA), and branch PAs (left PA [LPA] and right PA [RPA]) size by echocardiography before mBTS and surgical repair. Of 172 patients with TOF/PS, 40 (23%) were palliated with an mBTS at a median age of 23 days, and 31 met criteria for echocardiographic analysis. Palliated patients had hypoplastic PV, MPA, RPA, and LPA at baseline. All structures had significant absolute growth before surgical repair (p < 0.001). PV and MPA Z scores were unchanged, whereas branch PAs showed significant improvement (RPA, p = 0.03; LPA, p = 0.008). A PV-sparing repair was performed in 116 of 132 primary repairs (88%) and in 10 of 31 palliated patients (32%). At a median follow-up of 4.0 years (range, 0.7 to 12.6 years), no palliated patients required reintervention for right ventricular outflow tract obstruction. Significant absolute growth of the PV, MPA, RPA, and LPA occurs after an mBTS in neonates with TOF/PS. Branch PA size normalizes before complete repair. In one-third of patients with hypoplastic PVs, absolute growth after an mBTS may have facilitated valve-sparing repair.


Cloning and characterization of rhesus cytomegalovirus glycoprotein B.
Rhesus cytomegalovirus (RhCMV) infection of rhesus macaques is an important model to investigate critical issues of cytomegalovirus biology. To better understand host immunological responses to viral glycoproteins, the glycoprotein B (gB) gene of RhCMV was molecularly cloned, sequenced and characterized. Transcription analysis revealed that RhCMV gB was transcribed as a late gene. The RhCMV gB gene encoded a predicted protein of 854 amino acids that was 60% identical/75% similar to the human CMV (HCMV) gB protein. The region of HCMV gB proposed to be responsible for virus binding to host cells, fusion and cell-to-cell spread was the most highly conserved region with RhCMV gB (74% identity/85% similarity). Conserved elements included 11 of 12 cysteine residues, 14 of 16 potential N-linked glycosylation sites and cross-reactive epitopes. Metabolic labelling experiments demonstrated that RhCMV gB was proteolytically processed similarly to HCMV gB. These results are critical for investigating virus-host relationships in CMV-infected primates.


Predicting post-discharge complications in cardiothoracic surgery: A clinical decision support system to optimize remote patient monitoring resources.
Cardiac surgery patients are highly prone to severe complications post-discharge. Close follow-up through remote patient monitoring can help detect adverse outcomes earlier or prevent them, closing the gap between hospital and home care. However, equipment is limited due to economic and human resource constraints. This issue raises the need for efficient risk estimation to provide clinicians with insights into the potential benefit of remote monitoring for each patient. Standard models, such as the EuroSCORE, predict the mortality risk before the surgery. While these are used and validated in real settings, the models lack information collected during or following the surgery, determinant to predict adverse outcomes occurring further in the future. This paper proposes a Clinical Decision Support System based on Machine Learning to estimate the risk of severe complications within 90 days following cardiothoracic surgery discharge, an innovative objective underexplored in the literature. Health records from a cardiothoracic surgery department regarding 5 045 patients (60.8% male) collected throughout ten years were used to train predictive models. Clinicians' insights contributed to improving data preparation and extending traditional pipeline optimization techniques, addressing medical Artificial Intelligence requirements. Two separate test sets were used to evaluate the generalizability, one derived from a patient-grouped 70/30 split and another including all surgeries from the last available year. The achieved Area Under the Receiver Operating Characteristic curve on these test sets was 69.5% and 65.3%, respectively. Also, additional testing was implemented to simulate a real-world use case considering the weekly distribution of remote patient monitoring resources post-discharge. Compared to the random resource allocation, the selection of patients with respect to the outputs of the proposed model was proven beneficial, as it led to a higher number of high-risk patients receiving remote monitoring equipment.


Intensive diabetes therapy and glomerular filtration rate in type 1 diabetes.
An impaired glomerular filtration rate (GFR) leads to end-stage renal disease and increases the risks of cardiovascular disease and death. Persons with type 1 diabetes are at high risk for kidney disease, but there are no interventions that have been proved to prevent impairment of the GFR in this population. In the Diabetes Control and Complications Trial (DCCT), 1441 persons with type 1 diabetes were randomly assigned to 6.5 years of intensive diabetes therapy aimed at achieving near-normal glucose concentrations or to conventional diabetes therapy aimed at preventing hyperglycemic symptoms. Subsequently, 1375 participants were followed in the observational Epidemiology of Diabetes Interventions and Complications (EDIC) study. Serum creatinine levels were measured annually throughout the course of the two studies. The GFR was estimated with the use of the Chronic Kidney Disease Epidemiology Collaboration formula. We analyzed data from the two studies to determine the long-term effects of intensive diabetes therapy on the risk of impairment of the GFR, which was defined as an incident estimated GFR of less than 60 ml per minute per 1.73 m(2) of body-surface area at two consecutive study visits. Over a median follow-up period of 22 years in the combined studies, impairment of the GFR developed in 24 participants assigned to intensive therapy and in 46 assigned to conventional therapy (risk reduction with intensive therapy, 50%; 95% confidence interval, 18 to 69; P=0.006). Among these participants, end-stage renal disease developed in 8 participants in the intensive-therapy group and in 16 in the conventional-therapy group. As compared with conventional therapy, intensive therapy was associated with a reduction in the mean estimated GFR of 1.7 ml per minute per 1.73 m(2) during the DCCT study but during the EDIC study was associated with a slower rate of reduction in the GFR and an increase in the mean estimated GFR of 2.5 ml per minute per 1.73 m(2) (P<0.001 for both comparisons). The beneficial effect of intensive therapy on the risk of an impaired GFR was fully attenuated after adjustment for glycated hemoglobin levels or albumin excretion rates. The long-term risk of an impaired GFR was significantly lower among persons treated early in the course of type 1 diabetes with intensive diabetes therapy than among those treated with conventional diabetes therapy. (Funded by the National Institute of Diabetes and Digestive and Kidney Diseases and others; DCCT/EDIC ClinicalTrials.gov numbers, NCT00360815 and NCT00360893.).


Sex differences in fitness and cardiac function during exercise in adolescents with chronic fatigue.
Females demonstrate less robust Frank-Starling mechanism with respect to cardiac preload than males at rest. We asked whether this phenomenon would also affect cardiac performance during exercise. We hypothesized that stroke volume (SV) response to exercise would be more limited in deconditioned females such that cardiac output would be mainly rate dependent, compared with males. We conducted a chart audit of clinical exercise tests performed by adolescents with chronic fatigue. Oxygen uptake (V˙O2) was measured breath-by-breath at rest and during cycle ergometry, while cardiac output was measured by acetylene rebreathing at rest plus 2-3 subthreshold workloads. SV response was analyzed in two ways: after normalization for body surface area (SV index, SVI) and as percentage change from resting values. Among 304 adolescents (78% females) with chronic fatigue, 189 (80%) of 236 females and 52 (76%) of 68 males were deconditioned (peakV˙O2 <90% predicted). Heart rate trajectory during exercise was steeper for unfit than fit females, 70 vs 61 beat·min-1 per L·min-1 V˙O2, (P=.003); but not for males, 47 vs 42 beat·min-1 per L·min-1 V˙O2 (P=.23). The highest measured SVI did not differ between unfit vs fit females (42.8 vs 41.5 mL·m-2 , P=.39) while fit males showed larger SV during exercise than their unfit peers (highest SVI 55.9 vs 48.0 mL·m-2 , P=.014). Both qualitative and quantitative sex differences exist in SV responses to exercise among chronically fatigued adolescents, suggesting volume loading may be more efficacious in girls.


Differentiation of Tr1 cells by immature dendritic cells requires IL-10 but not CD25+CD4+ Tr cells.
Dendritic cells (DCs) are specialized antigen-presenting cells that monitor the antigenic environment and activate naive T cells. The role of DCs is not only to sense danger but also to tolerize the immune system to antigens encountered in the absence of maturation/inflammatory stimuli. Indeed, if a naive T cell encounters its antigen on immature DCs (iDCs), it may differentiate into a T-regulatory (Tr) rather than a T-effector cell. However, little is known about the mechanisms by which iDCs differentiate Tr cells. We developed a standardized and highly reproducible protocol to differentiate Tr cells by repetitive exposure of naive peripheral blood CD4(+) T cells to allogeneic iDCs. The resultant Tr cells are phenotypically and functionally identical to type 1 Tr (Tr1) cells because their generation requires production of IL-10 by iDCs, and they suppress T-cell responses through an interleukin-10 (IL-10)- and a transforming growth factor beta (TGF-beta)-dependent mechanism. In addition, Tr1 cells induced by iDCs do not require the presence of CD4(+)CD25(+) Tr cells for their generation, nor do they express high constitutive levels of CD25 or the transcription factor FoxP3. Thus, iDCs can drive the differentiation of Tr1 cells and can be used to generate large numbers of alloantigen-specific Tr1 cells for clinical use as a cellular therapy to restore peripheral tolerance.


The effect of sugars on (pro)insulin biosynthesis.
Rates of incorporation of [4,5-(3)H]leucine into insulin plus proinsulin, designated ;(pro)insulin', and total protein in rat pancreatic islets were measured. Glucose stimulates rates of total protein and (pro)insulin biosynthesis, but (pro)insulin biosynthesis is stimulated preferentially. Mannose and N-acetylglucosamine also stimulate (pro)insulin and total protein biosynthesis; inosine and dihydroxyacetone stimulate (pro)insulin biosynthesis specifically. Fructose does not stimulate (pro)insulin biosynthesis when tested alone, but does so in the presence of low concentrations of glucose, mannose or N-acetylglucosamine. Many glucose analogues do not stimulate (pro)insulin biosynthesis. Mannoheptulose inhibits synthesis of (pro)insulin and total protein stimulated by glucose or mannose but not by dihydroxyacetone, inosine or N-acetylglucosamine; phloretin (9mum) inhibits N-acetylglucosamine-stimulated (pro)insulin biosynthesis preferentially. The data are in agreement with the view that the same glucose-sensor mechanism may control both insulin release and biosynthesis, and ;substrate-site' model is suggested. The threshold for stimulation of biosynthesis of (pro)insulin and total protein is lower than that found for glucose-stimulated insulin release; moreover the biosynthetic response to an elevation of glucose concentration is slower than that found for insulin release. The physiological implication of these findings is discussed. Caffeine and isobutylmethylxanthine, at concentrations known to increase islet 3':5'-cyclic AMP and potentiate glucose-induced insulin release, were without effect on rates of glucose-stimulated (pro)insulin biosynthesis.


The prognostic value of preoperative inflammation-based prognostic scores and nutritional status for overall survival in resected patients with nonmetastatic Siewert type II/III adenocarcinoma of esophagogastric junction.
Immune and nutritional status of patients have been reported to predict postoperative complications, recurrence, and prognosis of patients with cancer. Therefore, this retrospective study aimed to explore the prognostic value of preoperative inflammation-based prognostic scores [neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR)] and nutritional status [prognostic nutritional index (PNI), body mass index (BMI), hemoglobin, albumin, and prealbumin] for overall survival (OS) in adenocarcinoma of esophagogastric junction (AEG) patients. A total of 355 patients diagnosed with Siewert type II/III AEG and underwent surgery between October 2010 and December 2011 were followed up until October 2016. Receiver operating characteristic (ROC) curve analysis was used to determine the cutoff values of NLR, PLR, and PNI. Kaplan-Meier curves and Cox regression analyses were used to calculate the OS characteristics. The ideal cutoff values for predicting OS were 3.5 for NLR, 171 for PLR, and 51.3 for PNI according to the ROC curve. The patients with hemoglobin <120 g/L (P = .001), prealbumin <180 mg/L (P = .000), PNI <51.3 (P = .010), NLR >3.5 (P = .000), PLR >171 (P = .006), and low BMI group (P = .000) had shorter OS. And multivariate survival analysis using the Cox proportional hazards model showed that the tumor-node-metastasis stage, BMI, NLR, and prealbumin levels were independent risk factors for the OS. Our study demonstrated that preoperative prealbumin, BMI, and NLR were independent prognostic factors of AEG patients.


Diagnostic accuracy of ultrasound in evaluation of obstructive jaundice with MRCP as gold standard.
To evaluate the diagnostic accuracy of ultrasound in obstructive jaundice taking MRCP as gold standard. This cross-sectional study was conducted at Dow Institute of Radiology (DIR), Dow University of Health Sciences (DUHS), Karachi from 2nd May 2018 till 2nd November 2018. Both male and female patients aged 30 to 80 years with suspected obstructive jaundice were included. Patients already diagnosed with obstructive jaundice were excluded. MRCP and ultrasound were performed in suspected patients. Diagnostic accuracy including sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of obstructive jaundice were calculated using contingency tables using MRCP findings as gold standard. Mean age of the patients was 54.73 ± 12.65 years. In causes of obstruction, choledocholothiasis was responsible for 85 (35.1%), stricture 61 (25.2%), carcinoma of head of pancreas 39 (16.1%), periampullary carcinoma 21 (8.7%), cholangiocarcinoma 10 (4.1%) and gallbladder carcinoma 26 (10.7%) of the cases. Diagnostic accuracy of ultrasound in obstructive jaundice taking MRCP findings as gold standard showed sensitivity, specificity, positive predicted value (PPV), negative predicted value (NPV) and overall diagnostic accuracy as 84.57%, 79.10%, 91.36%, 66.25% and 83.06%. Ultrasound has a high sensitivity, moderate specificity, and high diagnostic accuracy in diagnosis of obstructive jaundice.


Bioassay battery interlaboratory investigation of emerging contaminants in spiked water extracts - Towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring.
Bioassays are particularly useful tools to link the chemical and ecological assessments in water quality monitoring. Different methods cover a broad range of toxicity mechanisms in diverse organisms, and account for risks posed by non-target compounds and mixtures. Many tests are already applied in chemical and waste assessments, and stakeholders from the science-police interface have recommended their integration in regulatory water quality monitoring. Still, there is a need to address bioassay suitability to evaluate water samples containing emerging pollutants, which are a current priority in water quality monitoring. The presented interlaboratory study (ILS) verified whether a battery of miniaturized bioassays, conducted in 11 different laboratories following their own protocols, would produce comparable results when applied to evaluate blinded samples consisting of a pristine water extract spiked with four emerging pollutants as single chemicals or mixtures, i.e. triclosan, acridine, 17α-ethinylestradiol (EE2) and 3-nitrobenzanthrone (3-NBA). Assays evaluated effects on aquatic organisms from three different trophic levels (algae, daphnids, zebrafish embryos) and mechanism-specific effects using in vitro estrogenicity (ER-Luc, YES) and mutagenicity (Ames fluctuation) assays. The test battery presented complementary sensitivity and specificity to evaluate the different blinded water extract spikes. Aquatic organisms differed in terms of sensitivity to triclosan (algae > daphnids > fish) and acridine (fish > daphnids > algae) spikes, confirming the complementary role of the three taxa for water quality assessment. Estrogenicity and mutagenicity assays identified with high precision the respective mechanism-specific effects of spikes even when non-specific toxicity occurred in mixture. For estrogenicity, although differences were observed between assays and models, EE2 spike relative induction EC50 values were comparable to the literature, and E2/EE2 equivalency factors reliably reflected the sample content. In the Ames, strong revertant induction occurred following 3-NBA spike incubation with the TA98 strain, which was of lower magnitude after metabolic transformation and when compared to TA100. Differences in experimental protocols, model organisms, and data analysis can be sources of variation, indicating that respective harmonized standard procedures should be followed when implementing bioassays in water monitoring. Together with other ongoing activities for the validation of a basic bioassay battery, the present study is an important step towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring.


Foot skin ischaemia in atherosclerotic peripheral vascular disease.
Hyperbaric oxygen and the vasodilating effect of tolazoline hydrochloride were used to investigate atherosclerotic ischaemia of the skin of the foot. Ischaemic feet were divided into two subgroups each with a foot blood flow significantly higher than normal and significantly different from each other. The high blood flow in the ischaemic feet appears to have been an attempt to meet a tissue oxygen need. In some instances this need seems to have been satisfied but without obvious benefit to the ischaemic or anoxic skin. It is suggested that a local rather than a regional blood flow insufficiency is the cause of skin lesions in peripheral vascular disease.


Effects of variance in mini amplitude on stimulus-evoked release: a comparison of two models.
The strength of synaptic connections between two neurons is characterized by the number of release sites (N) on the presynaptic cell, the probability (p) of transmitter release at those sites in response to a stimulus, and the average size (A) of the postsynaptic response from each site. Quantal analysis can determine N, p, and A, but the large variance in the amplitudes of minis at central synapses is predicted to obscure quantal peaks and render quantal analysis unusable. Recently it has been suggested that the variance in mini amplitude is generated by differences between release sites, rather than by quantum-to-quantum fluctuations at identical sites, and that this form of variance in mini amplitude reduces the amount of variance expected in quantal peaks. Using simulations, we examine the possibility of resolving quantal peaks assuming either form of variance in mini amplitude. We find that individual quantal peaks are resolvable in neither case, provided that the uniquantal distribution is similar to the mini distribution. Because this lack of resolution compromises the utility of quantal analysis, we develop a general description that can solve N and p, given the statistical parameters of the mini distribution and the evoked distribution. We find that this description is relatively insensitive to the source of variance in mini amplitude.


Implementation of ward-based clinical pharmacy services in Belgium--description of the impact on a geriatric unit.
Patient-centered clinical pharmacy services are still poorly developed in Europe, despite their demonstrated advantages in North America and the UK. Reporting European pilot experiences is, therefore, important to assess the usefulness of clinical pharmacy services in this specific context. To report the results of the first implementation of Belgian clinical pharmacy services targeting patients at high risk of drug-related problems. An intervention study was conducted by a trained clinical pharmacist providing pharmaceutical care to 101 patients (mean age 82.2 y; mean +/- SD number of prescribed drugs 7.8 +/- 3.5) admitted to an acute geriatric unit, over a 7 month period. All interventions to optimize prescribing, and their acceptance, were recorded. An external panel (2 geriatricians, 1 clinical pharmacist) assessed the interventions' clinical significance. Persistence of interventions after discharge was assessed through telephone calls. A total of 1066 interventions were made over the 7 month period. The most frequent drug-related problems underlying interventions were: underuse (15.9%), wrong dose (11.9%), inappropriate duration of therapy (9.7%), and inappropriate choice of medicine (9.6%). The most prevalent consequences were to discontinue a drug (24.5%), add a drug (18.6%), and change dosage (13.7%). Acceptance rate by physicians was 87.8%. Among interventions with clinical impact, 68.3% and 28.6% had moderate and major clinical significance, respectively. Persistence of chronic treatment changes 3 months after discharge was 84%. Involving a trained clinical pharmacist in a geriatric team led to clinically relevant and well-accepted optimization of medicine use. This initiative may be a springboard for further development of clinical pharmacy services.


Characterization of insulin-like growth factor-I and its receptor and binding proteins in transected nerves and cultured Schwann cells.
The insulin-like growth factors (IGFs) are trophic factors whose growth-promoting actions are mediated via the IGF-I receptor and modulated by six IGF binding proteins (IGFBPs). In this study, we observed increased transcripts of both IGF-I and IGF-I receptor after rat sciatic nerve transection. Schwann cells (SCs) were the main source of IGF-I and IGFBP-5 immunoreactivity until 7 days after nerve transection, when invading macrophages in the distal nerve stumps were strongly IGF-I positive. In vitro, IGF-I promoted SC mitogenesis. Northern analysis revealed that SCs expressed IGF-I receptor and IGFBP-5. IGF-I treatment increased the intensity of IGFBP-5 without affecting gene expression. Des(1-3)IGF-I, an IGF-I analogue with low affinity for IGFBP, had no such effect. Incubation of recombinant human IGFBP-5 with SC conditioned media revealed IGF-I protection of IGFBP-5 from proteolysis, implying the presence of an IGFBP-5 protease in SC conditioned media. Collectively, these data support the concept that, in response to nerve injury, invading macrophages produce IGF-I and SC express the IGF-I receptor, to facilitate regeneration. This regenerative process may be augmented further by the ability of SC to secrete IGFBPs, which in turn may increase local IGF-I bioavailability.


Effects of 1 year of daily teriparatide treatment on iliacal bone mineralization density distribution (BMDD) in postmenopausal osteoporotic women previously treated with alendronate or risedronate.
Anabolic treatment with teriparatide of postmenopausal osteoporotic patients previously treated with bisphosphonates is a new therapeutic approach. However, its effects on the bone mineralization density distribution (BMDD) are unknown. We studied paired transiliac bone biopsy samples taken before and after 1 year of treatment with recombinant human parathyroid hormone peptide 1-34 (teriparatide) from 16 osteoporotic women treated with either alendronate (priorALN) or risedronate (priorRIS) for at least 2 years and subsequently treated for 12 months with teriparatide. Cancellous (Cn.) and cortical (Ct.) BMDD values were measured using quantitative backscattered electron imaging. At baseline, BMDD values of priorALN and priorRIS women were similar and within the normal range. One year of teriparatide treatment caused significant effects on the BMDD. Analyzing changes from baseline for each bisphosphonate group separately, priorALN patients revealed increases in the portion of low mineralized bone areas (Cn.Ca(Low) +25.9%, Ct.Ca(Low) +62.0%, both p < .05) and Ct. heterogeneity of mineralization (Ct.Ca(Width) +22.8%, p < .001). PriorRIS patients showed increased mineralization heterogeneity (Cn.Ca(Width) +14.8%, p < .05, and Ct.Ca(Width) +15.8%, p < .001). Analysis of the influence of the prior bisphosphonate treatment showed that the BMDD response to 1 year of teriparatide treatment did not depend on the type of prior bisphosphonate. In consequence, priorALN and priorRIS groups were combined. The pooled groups revealed increased Cn.Ca(Width) and Ct.Ca(Width) (+10.7%, p < .01, and +19.6%, p < .001, respectively) as well as increased Cn.Ca(Low) and Ct.Ca(Low) (+18.2%, p < .05, and +36.6%, p < .01, respectively). In summary, our findings indicate a significant effect of teriparatide on BMDD when administered subsequent to a bisphosphonate in agreement with teriparatide's anabolic action.


Relationship of IL-6, IL-8, TNF and sICAM-1 levels to PROM, pPROM, and the risk of early-onset neonatal sepsis.
Intraamniotic infections negatively affect the mortality and morbidity in parturients and newborns. The prognosis of the disease is associated with a timely diagnosis of these conditions. One of approaches to providing timely information on the risk of the initiation of intra-amniotic infection and early-onset neonatal sepsis is the examination of cytokine levels. The purpose of the work was to evaluate the importance of the cytokines, IL-6, IL-8, and TNF-alpha, and the adhesive molecule, sICAM-1, as risk factors for early-onset neonatal sepsis and intra-amniotic infections. In a group of 152 women we sampled the blood from the umbilical cord vein immediately after delivery for the determination of the cytokines IL-6, IL-8 and TNF-alpha, and the adhesive molecule, sICAM-1, in newborns. The sensitivity and specificity results are as follows, respectively: IL-6, 0.800 and 0.972; TNF-alpha, 0.364 and 0.943; IL-8, 0.875 and 0.965; and sICAM-1, 0.833 and 0.952. For screening purposes, it is suitable to determine levels of IL-8, IL-6, and sICAM-1. For the screening examination, one of the cytokines mentioned is sufficient, i.e., IL-8 or IL-6, or the level of the adhesive molecule, sICAM-1. It is unnecessary to combine these markers.


[Metabolism of vitamin D3 administered in liposomes in rat liver].
Incorporation of [3H]-cholecalciferol into the liver and the rate of its conversion into 25-OH-D are significantly larger after intravenous administration of this vitamin into liposomes than its dissolution in ethanol. The latter may be due to a different distribution of vitamin D3 in the cells of the liver. Incorporation of [3H]-vitamin D3 in reticulocytes suggests that its cells are a locus for storage of vitamin D, from which the latter is transported into hepatocytes where its 25-hydroxylation proceeds.


Urinary biopterin and neopterin excretion and pituitary-adrenal activity in psychiatric patients.
The urinary excretion of biopterin and neopterin, pterin compounds related to tetrahydrobiopterin, the cofactor for the initial steps in monoamine synthesis, was serially measured in a heterogeneous group of psychiatric patients and compared to excretion in control subjects, to state of illness, and to the results of the dexamethasone suppression test. Patients with major depression had increased excretion of biopterin compared to normal subjects. There was no relationship between biopterin or neopterin excretion and postdexamethasone cortisol levels. Pterin excretion did not significantly change with improvement in mood or with conversion from nonsuppressor to suppressor status. The meaning of increased urinary biopterin is presently unclear, although its relation to hormonal state and sympathoadrenal tone deserves further study.


Durable bonds at the adhesive/dentin interface: an impossible mission or simply a moving target?
Composite restorations have higher failure rates, more recurrent caries and increased frequency of replacement as compared to dental amalgam. Penetration of bacterial enzymes, oral fluids, and bacteria into the crevices between the tooth and composite undermines the restoration and leads to recurrent decay and failure. The gingival margin of composite restora tions is particularly vulnerable to decay and at this margin, the adhesive and its seal to dentin provides the primary barrier between the prepared tooth and the environment. The intent of this article is to examine physico-chemical factors that affect the integrity and durability of the adhesive/dentin interfacial bond; and to explore how these factors act synergistically with mechanical forces to undermine the composite restoration. The article will examine the various avenues that have been pursued to address these problems and it will explore how alterations in material chemistry could address the detrimental impact of physico-chemical stresses on the bond formed at the adhesive/dentin interface.


Increased Glycolysis and Higher Lactate Production in Hyperglycemic Myotubes.
Previous studies have shown that chronic hyperglycemia impairs glucose and fatty acid oxidation in cultured human myotubes. To further study the hyperglycemia-induced suppression of oxidation, lactate oxidation, mitochondrial function and glycolytic rate were evaluated. Further, we examined the intracellular content of reactive oxygen species (ROS), production of lactate and conducted pathway-ANOVA analysis on microarray data. In addition, the roles of the pentose phosphate pathway (PPP) and the hexosamine pathway were evaluated. Lactic acid oxidation was suppressed in hyperglycemic versus normoglycaemic myotubes. No changes in mitochondrial function or ROS concentration were observed. Pathway-ANOVA analysis indicated several upregulated pathways in hyperglycemic cells, including glycolysis and PPP. Functional studies showed that glycolysis and lactate production were higher in hyperglycemic than normoglycaemic cells. However, there were no indications of involvement of PPP or the hexosamine pathway. In conclusion, hyperglycemia reduced substrate oxidation while increasing glycolysis and lactate production in cultured human myotubes.


Have drug combinations supplanted stem cell transplantation in myeloma?
The introduction of proteasome inhibitor and immunomodulatory drugs has considerably changed the treatment paradigm of multiple myeloma. Autologous stem cell transplantation (ASCT) is superior to conventional chemotherapy and is considered the standard of care for patients younger than 65 years. Nevertheless, the favorable results shown by multidrug inductions, consolidations, and long-term maintenance approaches have challenged the role of ASCT. This article provides an overview of recent and ongoing clinical trials and aims to define the role of ASCT in the era of novel agents.


Breast cancer risk associated with benign breast disease: systematic review and meta-analysis.
Benign breast disease (BBD) is a broad category of diagnoses reported to convey a variable degree of increased risk of developing breast cancer. A meta-analysis of the existing literature was performed to quantify the risk estimate associated with BBD. Pubmed, Google Scholar, and EMBASE databases were searched in January 2011. English retrospective and prospective observational studies published from 1972 to 2010 evaluating BBD and the risk of breast cancer were included with data acquisition reported from 1930 to 2007. Eligibility was performed independently following a standardized protocol for full-text publication review by a single reviewer and reviewed by a second author. Of the 3,409 articles retrieved from the literature search, 32 studies met the selection criteria. Reported risk estimates, including relative risk, odds ratio, standardized incidence ratios, rate ratio, hazards ratio, and incidence rate ratio, were the primary outcomes extracted. The most commonly reported pathologies were decided prior to extraction and organized into the following categories for analysis of the extracted risk estimate: non-proliferative disease (NPD), proliferative disease without atypia, benign breast disease not otherwise specified (BBD), and atypical hyperplasia not otherwise specified (AHNOS). The mean age at benign breast biopsy was 46.1 years and the mean age of developing breast cancer was 55.9 years. The mean follow-up length was 12.8 years (range 3.3-20.6). The summary risk estimate of developing breast cancer for NPD was 1.17 (N = 8; 95% CI 0.94-1.47). Proliferative disease without atypia was associated with significantly increased risk of future breast cancer, summary relative risk 1.76 (N = 15; 95% CI 1.58-1.95). The summary risk estimate for AHNOS was 3.93 (N = 13; 95% CI 3.24-4.76). This meta-analysis demonstrates that proliferative benign breast disease with or without atypia is associated with a significant increase in risk of developing breast cancer. These data support management strategies for women with benign breast disease such as additional screening methods or chemoprevention.


Oral health conditions and cognitive functioning in middle and later adulthood.
The purpose of the present study was to examine the impact of oral health conditions on cognitive functioning on basis of data samples from several European countries. Secondary analyses were conducted of data from wave 2 of the Survey of Health, Ageing, and Retirement in Europe (SHARE) which includes 14 European countries and is intended to be representative of each country's middle and later adulthood population. Information on word recall, verbal fluency, and numeracy as well as information on chewing ability and denture wearing status was available for a total of 28,693 persons aged 50+. Multivariate regression analysis was used to detect influences of oral health parameters on cognitive functioning (p < 0.05). Persons with good chewing ability or without dentures had significantly better word recall, verbal fluency, and numeracy skills than persons with chewing impairment or with dentures. The observed patterns of parameter estimates imply differential oral health impacts on numeracy compared to word recall and verbal fluency. The present study provides novel large-scale epidemiological evidence supportive of an association between oral health and cognitive functioning. Future research should intend to verify the precise causal links between oral health conditions, various cognitive dimensions, and their neural correlates.


Assay formats involving acridinium-ester-labeled DNA probes.
We describe the development of several hybridization assay formats involving acridinium-ester-labeled DNA probes. The simplest of these is a homogeneous assay procedure that requires only three steps to complete, including a 5-s detection step. Using this format, we have detected target sequences in the 10(-16) to 10(-17) mol range; when rRNA is the target, this translates to 3000 to 300 bacterial organisms. The entire assay can be carried out in less than 30 min. This is the first homogeneous DNA probe assay to be of practical use in the clinical laboratory, and it represents a major simplification of hybridization formats. We also demonstrate the use of this homogeneous assay format to discriminate single-base differences between two closely related target sequences and to detect DNA as well as RNA target molecules. By combining homogeneous hybrid discrimination with solid-phase separation, we have been able to decrease background readings from unhybridized probe to only a few parts per million. This enhances assay sensitivity about 10-fold, to a range of 10(-17) to 10(-18) mol of target. We are in the process of further improving the performance of these assays.


Renal replacement therapy for rare diseases affecting the kidney: an analysis of the ERA-EDTA Registry.
In recent years, increased efforts have been undertaken to address the needs of patients with rare diseases by international initiatives and consortia devoted to rare disease research and management. However, information on the overall prevalence of rare diseases within the end-stage renal disease (ESRD) population is limited. The aims of this study were (i) to identify those rare diseases within the ERA-EDTA Registry for which renal replacement therapy (RRT) is being provided and (ii) to determine the prevalence and incidence of RRT for ESRD due to rare diseases, both overall and separately for children and adults. The Orphanet classification of rare disease was searched for rare diseases potentially causing ESRD, and these diagnosis codes were mapped to the corresponding ERA-EDTA primary renal disease codes. Thirty-one diagnoses were defined as rare diseases causing ESRD. From 1 January 2007 to 31 December 2011, 7194 patients started RRT for a rare disease (10.6% children). While some diseases were exclusively found in adults (e.g. Fabry disease), primary oxalosis, cystinosis, congenital anomalies of the kidney and urinary tract (CAKUT) and medullary cystic kidney disease affected young patients in up to 46%. On 31 December 2011, 20 595 patients (12.4% of the total RRT population) were on RRT for ESRD caused by a rare disease. The point prevalence was 32.5 per million age-related population in children and 152.0 in adults. Only 5.8% of these patients were younger than 20 years; however, 57.7% of all children on RRT had a rare disease, compared with only 11.9% in adults. CAKUT and focal segmental glomerulosclerosis were the most prevalent rare disease entities among patients on RRT. More than half of all children and one of nine adults on RRT in the ERA-EDTA Registry suffer from kidney failure due to a rare disease, potentially with a large number of additional undiagnosed or miscoded cases. Comprehensive diagnostic assessment and the application of accurate disease classification systems are essential for improving the identification and management of patients with rare kidney diseases.


The cachectic mediator proteolysis inducing factor activates NF-kappaB and STAT3 in human Kupffer cells and monocytes.
A novel proteoglycan, proteolysis inducing factor (PIF), is capable of inducing muscle proteolysis during the process of cancer cachexia, and of inducing an acute phase response in human hepatocytes. We investigated whether PIF is able to activate pro-inflammatory pathways in human Kupffer cells, the resident macrophages of the liver, and in monocytes, resulting in the production of pro-inflammatory cytokines. Normal liver tissue was obtained from patients undergoing partial hepatectomy and Kupffer cells were isolated. Monocytes were isolated from peripheral blood. Following exposure to native PIF, pro-inflammatory cytokine production from Kupffer cells and monocytes was measured and the NF-kappaB and STAT3 transcriptional pathways were investigated using electrophoretic mobility shift assays. We demonstrate that PIF is able to activate the transcription factor NF-kappaB and NF-kappaB-inducible genes in human Kupffer cells, and in monocytes, resulting in the production of pro-inflammatory cytokines such as TNF-alpha, IL-8 and IL-6. PIF enhances the expression of the cell surface molecules LFA-1 and CD14 on macrophages. PIF also activates the transcription factor STAT3 in Kupffer cells. The pro-inflammatory effects of PIF, mediated via NF-kappaB and STAT3, are important in macrophage behaviour and may contribute to the inflammatory pro-cachectic process in the liver.


Delayed feedback as a means of control of noise-induced motion.
Time-delayed feedback is exploited for controlling noise-induced motion in coherence resonance oscillators. Namely, under the proper choice of time delay, one can either increase or decrease the regularity of motion. It is shown that in an excitable system, delayed feedback can stabilize the frequency of oscillations against variation of noise strength. Also, for fixed noise intensity, the phenomenon of entrainment of the basic oscillation period by the delayed feedback occurs. This allows one to steer the time scales of noise-induced motion by changing the time delay.


Biliary metal stents and air embolism: a note of caution.
Metal stents are a valuable treatment modality for patients with biliary obstruction. However, we present here two patients whose cases may serve as a warning about an unusual complication associated with these stents. We encountered this complication after endoscopic retrograde cholangiography for obstructed metal biliary stents. The first patient, an 87-year-old man with a benign biliary stricture, failed to regain consciousness after clearing of his stent using a Dormia basket and balloon catheter. Cerebral air embolism was diagnosed on cerebral computed tomography, and transesophageal echocardiography revealed a patent foramen ovale as a precipitating factor for paradoxical air embolism. He survived and was discharged with a residual hemiparesis. In the second patient, a 54-year-old man who had a history of a Billroth II operation and chronic pancreatitis and who had a portal cavernoma with biliary obstruction due to collateral veins, electromechanical dissociation complicated the balloon-catheter stent revision. Echocardiography performed during cardiopulmonary resuscitation showed major air embolism to the right heart. The patient died. These cases demonstrate that air may gain access to the venous system during therapeutic endoscopic procedures of this type. It is likely that the large diameter of metal stents and the potential for these stents to lacerate venous structures facilitate the entry of air into the venous circulation, an event which may have life-threatening consequences.


Characteristics of the desensitization and resensitization of the cyclic AMP-independent glycogenolytic response in rat liver cells.
Vasopressin and alpha-adrenergic agonists were shown previously [Bréant, Keppens & De Wulf (1981) Biochem. J. 200, 509-514] to induce a heterologous, dose-dependent and receptor-mediated desensitization of the cyclic AMP-independent glycogenolytic response in isolated hepatocytes. The desensitized state of the hepatocytes can be preserved as long as the agonist is bound to its receptor. Conversely, washing the cells with a hormone-free buffer or displacement of the agonist from its receptor by a specific antagonist restores the responsiveness. The desensitization and its reversibility (i.e. resensitization) are obtained within minutes. The desensitization can be clearly elicited at temperatures as low as 5 degrees C, whereas the glycogenolytic response and the enhancement of the 45Ca flux are only obtained above 15 degrees C; the resensitization requires even higher temperatures. A tentative model is proposed to account for the observed effects.


The influence of freshwater inflows on spawning success and early growth of an estuarine resident fish species, Acanthopagrus butcheri.
The influence of freshwater inflows and salinity on spawning success of black bream Acanthopagrus butcheri (Sparidae) was investigated over 2 years in a small estuary on the east coast of Tasmania, Australia. The individual spawning seasons experienced quite different freshwater inflows; 2004-2005 was characterized by low flows throughout the season whereas during 2005-2006 there were three relatively large discharge events in the first part of the season. Macroscopic gonad staging of adults was used to define the spawning season and daily increment analysis of otoliths from recently settled recruits was used to backcalculate spawning dates. Gonad staging indicated that adults were in spawning condition over a 3 to 4 month period during spring and summer. The timing and duration of successful spawning, however, differed markedly between years and was linked to the timing of freshwater inflows and salinity conditions, with successful spawning occurring during periods of low freshwater discharge and when salinities in the upper estuary were above c. 15. Growth rates of the recently settled recruits did not differ between years, nor did the timing of spawning within the season influence growth rates. While the latter finding was unexpected, especially given within season temperature variability, these results imply that by the onset of winter earlier spawned fish would be larger than later spawned individuals, potentially conferring advantages for survival and competition for food. Climate change predictions for eastern Tasmania indicate a decrease in river flows in spring and an increase during summer, potentially increasing environmental variability between and within years, with implications for spawning success and subsequent recruitment.


Rare copy number variants in tourette syndrome disrupt genes in histaminergic pathways and overlap with autism.
Studies of copy number variation (CNV) have characterized loci and molecular pathways in a range of neuropsychiatric conditions. We analyzed rare CNVs in Tourette syndrome (TS) to identify novel risk regions and relevant pathways, to evaluate burden of structural variation in cases versus controls, and to assess overlap of identified variations with those in other neuropsychiatric syndromes. We conducted a case-control study of 460 individuals with TS, including 148 parent-child trios and 1131 controls. CNV analysis was undertaken using 370 K to 1 M probe arrays, and genotyping data were used to match cases and controls for ancestry. CNVs present in < 1% of the population were evaluated. While there was no significant increase in the number of de novo or transmitted rare CNVs in cases versus controls, pathway analysis using multiple algorithms showed enrichment of genes within histamine receptor (subtypes 1 and 2) signaling pathways (p = 5.8 × 10(-4) - 1.6 × 10(-2)), as well as axon guidance, cell adhesion, nervous system development, and synaptic structure and function processes. Genes mapping within rare CNVs in TS showed significant overlap with those previously identified in autism spectrum disorders but not intellectual disability or schizophrenia. Three large, likely pathogenic, de novo events were identified, including one disrupting multiple gamma-aminobutyric acid receptor genes. We identify further evidence supporting recent findings regarding the involvement of histaminergic and gamma-aminobutyric acidergic mechanisms in the etiology of TS and show an overlap of rare CNVs in TS and autism spectrum disorders.


Automatic reactor for solid-phase synthesis of molecularly imprinted polymeric nanoparticles (MIP NPs) in water.
We report the development of an automated chemical reactor for solid-phase synthesis of MIP NPs in water. Operational parameters are under computer control, requiring minimal operator intervention. In this study, "ready for use" MIP NPs with sub-nanomolar affinity are prepared against pepsin A, trypsin and α-amylase in only 4 hours.


Phylogenic analysis of Chinese Leishmania isolates based on small subunit ribosomal RNA (SSU rRNA) and 7 spliced leader RNA (7SL RNA).
The leishmaniases are zoonotic diseases caused by protozoan parasites of the genus Leishmania. Leishmaniases are still endemic in China, especially in the west and northwest froniter regions. To revalue the preliminary phylogenetic results of Chinese Leishmania isolates, we amplified partial fragment of small subunit ribosomal RNA (SSU rRNA) and 7 spliced leader RNA (7SL RNA), then tested the phylogenetic relationships among Chinese Leishmania isolates and their relatives by analyzing SSU rRNA gene sequences and 7SL RNA gene sequences. 19 SSU RNA sequences and 9 7SL RNA sequences were obtained in our study, then analyzed with 42 SSU RNA sequences and 32 7SL RNA sequences retrieved from Genbank, respectively. In the Bayesian analysis of the SSU RNA gene, the isolate MHOM/CN/93/GS7 and the isolate IPHL/CN/77/XJ771 are members of Leishmania donovani complex, while the isolate MHOM/CN/84/JS1 clustered with Leishmania tropica. The other 11 Chinese Leishmania isolates (MHOM/CN/90/WC, MCAN/CN/90/SC11, MHOM/CN/80/XJ801, MHOM/CN/85/GS4, MHOM/CN/84/SD1, MCAN/CN/86/SC7, MHOM/CN/54/#3, MHOM/CN/83/GS2, MHOM/CN/90/SC10H2, MHOM/CN/89/GS6 and MHOM/CN/ 89/GS5) form an unclassified group, defined as Leishmania sp., and the most relative species to this group is L. tarentolae. In the Bayesian analysis of the 7SL RNA gene, 9 Chinese Leishmania isolates also formed an unclassified group with L. tarentolae, including canine isolate 10, MHOM/CN/85/GS4, MHOM/CN/84/SD1, MCAN/CN/86/SC7, MHOM/CN/54/#3, MHOM/ CN/83/GS2, MHOM/CN/90/SC10H2, MHOM/CN/89/GS6 and MHOM/CN/89/GS5. We concluded that: (1) Chinese Leishmania isolates are non-monophyly group; (2) an unclassified group may exist in China, and the most relative species to this group is L. tarentolae; (3) MHOM/CN/84/JS1, which was previously assigned as L. donovani, was most genetically related to L. tropica strain MHOM/SU/74/K27.


Complete mitochondrial genome of the mulberry white caterpillar Rondotia menciana (Lepidoptera: Bombycidae).
The mulberry white caterpillar, Rondotia menciana, belongs to the lepidopteran family Bombycidae, in which the domestic silkworm, Bombyx mori is included. In this study, we describe the complete mitochondrial genome of R. menciana in terms of general genomic features and characteristic features found in the A+T-rich region. The 15,364 bp long genome consisted of a typical set of genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and 1 major non-coding A+T-rich region, with the typical arrangement found in Lepidoptera. Twelve of the 13 PCGs started with typical ATN codons, except for the COI, which began with CGA and twelve of 13 PCGs had complete stop codons, except for the COII, which ended with a single T. The 360 bp long A+T-rich region harbored the conserved sequence blocks typically found in lepidopteran insects. Additionally, the A+T-rich region of R. menciana contained one tRNA(Met)-like structure, which had a proper anticodon and secondary structure.


Environment and human exposure to persistent organic pollutants (POPs) in India: a systematic review of recent and historical data.
Persistent organic pollutants (POPs) have been used in a wide range of agricultural and industrial commodities, resulting in vigorous deterioration of environment and human health. A number of studies on the occurrence of POPs confirm their presence in various environmental compartments and human body. In order to deal with this global concern, India has recently prepared the National Implementation Plan (NIP) of the Stockholm Convention. Common beliefs point at India as a hot spot of POP contamination and human exposure; however no systematic analysis was ever performed so far considering all available past data on POP occurrence. This review aims to examine the distribution pattern of POPs in multicompartment environment and human samples, meta-analysis of time trends in exposure levels to environment and humans, and cross country comparison of POP contamination with China. Based on this review, it can be concluded that the Indian environment and human population are highly contaminated by DDTs and HCHs; however scarcity of data on other POPs makes it challenging to assess their nationwide human and environmental exposure. No evidence of a general decline in DDT and HCH residues in the environment and human body come out from the meta-analysis of time trend. While comparing contamination levels between India and China, tendency towards decline in POP contamination is visible in China, unlike India.


Changes in the nutritional status of school children and adolescents in Abeokuta, Nigeria between 1983 and 2006.
The prevalence of overweight and obesity among children is increasing worldwide with significant health and social consequences. The aim of this study was to determine the current nutritional status and its changes between 1983 and 2006 among school children and adolescents in a south western Nigeria town. five hundred and seventy children aged 5 to 19 years from randomly selected primary and secondary schools in Abeokuta, Nigeria had their weights and height measured using standard techniques. The data obtained were compared with data obtained from a similar cross sectional survey carried out in 1983. The National Centre for Health Statistics/World Health Organisation guidelines and cut-off points were used to determine the degree of underweight and stunting. Obesity prevalence was assessed using weight for age greater than 120 % of expected. The mean age of the participants was 12.2 + 3.41 years, 296 (51.5 %) were males. The mean Weight-for-Age Z -score and Height-for-Age Z-score were -1.2773 + 1.14 and -0.8569 + 1.19 respectively. The prevalence rates of underweight and stunting were 24.7 % and 17.4 % respectively. Using weight greater than 120 % of expected weight for age, five (1.7 %) male and 7 (2.6 %) female children were obese compared to 3.3 % male and 5.1 % female children who were obese in 1983. The prevalence of overweight and obesity has remained relatively unchanged between 1983 and 2006. However,under-nutrition remains a major nutritional problem among school children and adolescents in Abeokuta, Nigeria.


AT(2) receptor and tissue injury: therapeutic implications.
The renin-angiotensin system (RAS) plays an important role in the initiation and progression of tissue injuries in the cardiovascular and nervous systems. The detrimental actions of the AT1 receptor (AT1R) in hypertension and vascular injury, myocardial infarction and brain ischemia are well established. In the past twenty years, protective actions of the RAS, not only in the cardiovascular, but also in the nervous system, have been demonstrated. The so-called protective arm of the RAS includes AT2-receptors and Mas receptors (AT2R and MasR) and is characterized by effects different from and often opposing those of the AT1R. These include anti-inflammation, anti-fibrosis, anti-apoptosis and neuroregeneration that can counterbalance pathological processes and enable recovery from disease. The recent development of novel, small-molecule AT2R agonists offers a therapeutic potential in humans with a variety of clinical indications.


Epilepsy-associated levels of perceived stigma, their associations with treatment, and related factors: A cross-sectional study in urban and rural areas in Ecuador.
Epilepsy is felt to be a stigmatizing condition. Stigma has been considered one of the major factors contributing to the burden of epilepsy and to the treatment gap. Stigma has a negative effect on the management of people with epilepsy (PWE). Furthermore, lack of information and inappropriate beliefs are still the factors that most contribute to stigma and discrimination. In this study, we assessed the level of perceived stigma in urban and rural areas and we report their association with in antiepileptic drug (AED) use, effects on seeking medical care, and stigma-associated factors. A cross-sectional study in urban and rural areas in Ecuador from January 2015 until May 2016. People with a confirmed diagnosis of epilepsy were included using three sources of information. The survey was implemented through a questionnaire to determine perceived stigma and evaluate the factors associated. The perceived stigma was measured using the revised Jacoby's stigma scale to detect differences in levels of stigmatization. Access to treatment was evaluated through self-report of AED use, and attainment of medical care and stigma-associated factors were assessed. Furthermore, a multivariate analysis adjusted for possible confounders was performed using stigma as the outcome variable. A total of 243 PWE were interviewed, 65.8% reported feeling stigmatized and 39.1% reported a high stigmatized level. We found a significant difference in high stigma perception in the urban area compared to the rural area. However, the lack of use of AEDs was significantly higher in the rural areas. No significant correlation was found between use of AEDs and the levels of perceived stigma. PWE who did not talk about their condition and those who did not feel well informed about their epilepsy had significantly higher perceived stigma levels. Additionally, the multivariate analysis demonstrated that area, educational level, type of seizure, talk about epilepsy, and information were associated with perceived stigma. The stigma perception was relevant in all PWE. We found a higher stigma level perception in the urban compared to rural area. Moreover, the lack of treatment was a serious problem mainly in rural areas. Even though we did not find that perceived stigma was associated with AED use, our study pointed out the influence of educational level and information related to stigmatization. Consequently, a coordinated effort to reduce stigma should include strategies focused on PWE education and information about their condition.


Use of direct near-infrared fluorescent lymphography for thoracoscopic thoracic duct identification in 15 dogs with chylothorax.
To describe and assess techniques of intraoperative near-infrared fluorescence lymphography (NIRFL) using indocyanine green (ICG) for lymphatic duct identification during thoracoscopic thoracic duct (TD) ligation in dogs. Retrospective case series. Dogs (n = 15) with naturally occurring chylothorax that underwent TD ligation. Medical records of dogs treated with thoracoscopic TD ligation in which NIRFL was utilized for intraoperative TD identification were reviewed. Data retrieved included CT lymphography (CTL) and surgical data, fluorophore dose and injection site, and timing and quality of operative TD identification. Preoperative CTL was successful in 13/15 dogs and operative NIRFL was successful in 15/15 dogs. Popliteal lymph node injection achieved successful NIRFL within ≤10 minutes in 7/11 in which it was attempted. TDs identified by NIRFL imaging correlated with TDs identified by preoperative CTL in 12/13 cases in which CTL was achieved. In 1/13 cases, NIRFL identified small lymphatics not visible on CTL. In 5/9 cases in which methylene blue (MB) and ICG were combined for operative lymphography, no ducts or fewer ducts were recorded as identifiable by visible MB than by NIRFL or CTL. The median dose of ICG to achieve NIRFL imaging was 0.05 mg/kg. Thoracoscopic TD ligation was successfully achieved in all patients. NIRFL is a viable technique to aid in TD identification during thoracoscopy. High contrast NIRF illumination of the TD can be achieved with successful intraparenchymal injection of ICG into the popliteal lymph node.


Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids.
Polyneuropathy, hearing loss, ataxia, retinitis pigmentosa, and cataract (PHARC) is a rare genetic human neurological disorder caused by null mutations to the Abhd12 gene, which encodes the integral membrane serine hydrolase enzyme ABHD12. Although the role that ABHD12 plays in PHARC is understood, the thorough biochemical characterization of ABHD12 is lacking. Here, we report the facile synthesis of mono-1-(fatty)acyl-glycerol lipids of varying chain lengths and unsaturation and use this lipid substrate library to biochemically characterize recombinant mammalian ABHD12. The substrate profiling study for ABHD12 suggested that this enzyme requires glycosylation for optimal activity and that it has a strong preference for very-long-chain lipid substrates. We further validated this substrate profile against brain membrane lysates generated from WT and ABHD12 knockout mice. Finally, using cellular organelle fractionation and immunofluorescence assays, we show that mammalian ABHD12 is enriched on the endoplasmic reticulum membrane, where most of the very-long-chain fatty acids are biosynthesized in cells. Taken together, our findings provide a biochemical explanation for why very-long-chain lipids (such as lysophosphatidylserine lipids) accumulate in the brains of ABHD12 knockout mice, which is a murine model of PHARC.