Vulnerability of rabbit atrium to reentry by hypoxia. Role of inhomogeneity in conduction and wavelength.
In isolated superfused left atria of the rabbit, the inducibility of tachyarrhythmias by single early premature stimuli was highly increased by hypoxia. High-resolution mapping showed that these arrhythmias were caused by circus movement around a functional arc of conduction block (leading circle reentry). To determine the electrophysiological changes by hypoxia responsible for the higher vulnerability to intra-atrial reentry, the wavelength of the atrial impulse and spatial inhomogeneities in refractory periods and local conduction delays were measured. Hypoxia caused a transient increase in refractory periods during the first 10-15 min of hypoxia. After this period, refractory periods shortened again to values slightly lower than during control. During the whole period of hypoxia, local differences in refractory periods were enlarged. Conduction velocity was significantly depressed by hypoxia. As a result, the wavelength of the atrial impulse gradually shortened during hypoxia to approximately 80% of control. Inhomogeneity in conduction was quantified by phase maps in which the maximal local delays in conduction are plotted. Hypoxia caused a marked increase in inhomogeneity in conduction both during slow rhythm (inhomogeneity index increased from 2.3 to 3.4) and premature activation (from 3.1 to 4.7). We conclude that the higher vulnerability of the atrium for reentrant arrhythmias by hypoxia is based on a combination of a moderate shortening of the wavelength and an increase in inhomogeneity in conduction of premature wavefronts.


Reactivity of monoclonal antibody OKM5 with sebaceous carcinoma.
Two cases of sebaceous carcinoma (SC), as well as epithelial tumours, melanoma, and lymphoma, were examined using immunoperoxidase and a panel of monoclonal antibodies on cryostat sections. The results showed that, whereas all SC cells in both cases reacted strongly with monoclonal antibody OKM5, other tumour cells (except juvenile xanthogranuloma cells) did not. The pagetoid cells within the epidermis of SC also reacted with OKM5 antibody. Although the nature of the phenomenon merits further study, this reactivity, or cross-reactivity, might possibly aid diagnosis of SC.


Accuracy of angiographic determination of left main coronary arterial narrowing. Angiographic--histologic correlative analysis in 28 patients.
To evaluate the accuracy of coronary angiography in identifying severe narrowing of the left main coronary artery (LMCA), the degree of narrowing observed by angiography was compared to that observed at necropsy in 28 patients with symptomatic coronary heart disease in whom angiography had been performed within 40 days of death. The angiograms were evaluated independently by three experienced angiographers. In 20 of the 28 patients (71%), the degree of narrowing of the LMCA was either underestimated (13 patients) or overestimated (10 patients) by two or three of three angiographers; of 84 angiographic judgments made by the three angiographers in the 28 patients, 54 (64%) were underestimates (33 judgments, 39%) or overestimates (21 judgments, 25%) of the degree of LMCA narrowing. Of 12 LMCAs narrowed 76-100% in cross-sectional area at necropsy, six were underestimated at preoperative angiography by two or three of three angiographers; of 12 LMCAs narrowed 51-75% in cross-sectional area at necropsy, all 12 were either under- or overestimated angiographically by two or three of three angiographers; of four LMCAs narrowed 26-50% in cross-section at necropsy, two were overestimated by two of three angiographers. Thus, angiographic determination of degrees of narrowing of the LMCA during life is subject to considerable error. The angiographic errors appear to have resulted primarily from the presence of atherosclerotic plaque in the LMCA and an insufficient number of angiographic projections.


Geriatric rehabilitation.
Elderly people experience motor, sensory and cognitive losses that affect their mobility and their ability to perform activities of daily living. There is often gradual loss of function, which requires recognition and correction. Sudden events, such as stroke, amputation or trauma, may occur in an environment of diminished function and reserve. Rehabilitation measures must be directed toward prevention of dysmobility, correction of functional losses and provision for continuing aftercare.


Child Language Data Exchange System Tools for Clinical Analysis.
The Child Language Data Exchange System Project has developed methods for analyzing many aspects of child language development, including grammar, lexicon, discourse, gesture, phonology, and fluency. This article will describe the methods available for each of these six fields, and how they can be used for assessment in the clinical setting.


Joint aspiration and injection and synovial fluid analysis.
Joint aspiration/injection and synovial fluid (SF) analysis are both invaluable procedures for the diagnosis and treatment of joint disease. This chapter addresses: (1) the indications, the technical principles and the expected benefits and risks of aspiration and injection of intra-articular corticosteroid; and (2) practical aspects relating to SF analysis, especially in relation to crystal identification. Intra-articular injection of long-acting insoluble corticosteroids is a well-established procedure that produces rapid pain relief and resolution of inflammation in most injected joints. The knee is the most common site to require aspiration, although any non-axial joint is accessible for obtaining SF. The technique requires a knowledge of basic anatomy and should not be unduly painful for the patient. Provided sterile equipment and a sensible, aseptic approach are used, it is very safe. Analysis of aspirated SF is helpful in the differential diagnosis of arthritis and is the definitive method for diagnosis of septic arthritis and crystal arthritis. The gross appearance of SF can provide useful diagnostic information in terms of the degree of joint inflammation and presence of haemarthrosis. Microbiological studies of SF are the key to the confirmation of infectious conditions. Increasing joint inflammation is associated with increased SF volume, reduced viscosity, increasing turbidity and cell count, and increasing ratio of polymorphonuclear: mononuclear cells, but such changes are non-specific and must be interpreted in the clinical setting. However, detection of SF monosodium urate and calcium pyrophosphate dihydrate crystals, even from un-inflamed joints during intercritical periods, allow a precise diagnosis of gout and of calcium pyrophosphate crystal-related arthritis.


Severity Detection for the Coronavirus Disease 2019 (COVID-19) Patients Using a Machine Learning Model Based on the Blood and Urine Tests.
The recent outbreak of the coronavirus disease-2019 (COVID-19) caused serious challenges to the human society in China and across the world. COVID-19 induced pneumonia in human hosts and carried a highly inter-person contagiousness. The COVID-19 patients may carry severe symptoms, and some of them may even die of major organ failures. This study utilized the machine learning algorithms to build the COVID-19 severeness detection model. Support vector machine (SVM) demonstrated a promising detection accuracy after 32 features were detected to be significantly associated with the COVID-19 severeness. These 32 features were further screened for inter-feature redundancies. The final SVM model was trained using 28 features and achieved the overall accuracy 0.8148. This work may facilitate the risk estimation of whether the COVID-19 patients would develop the severe symptoms. The 28 COVID-19 severeness associated biomarkers may also be investigated for their underlining mechanisms how they were involved in the COVID-19 infections.


Profiles of depressive symptoms in older adults diagnosed with major depression: latent cluster analysis.
To explore the underlying structure of symptom presentation in older adults with major depression by identifying homogeneous clusters of individuals based on symptom profiles. Secondary data analysis using latent class cluster analysis. Clinical Research Center for the Study of Depression in Later Life conducted at Duke University. Three hundred sixty-six patients age 60+ who met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria for major depression and were enrolled in a longitudinal naturalistic treatment study. Responses to the 10 items of the Montgomery-Asberg Depression Rating Scale at the time of study enrollment. The authors identified four latent clusters of older adults with major depression. Patients in Cluster 1 (47.2%) had mean scores of average severity for reported and apparent sadness and lassitude and low mean scores for reduced appetite. Patients in Cluster 2 (27.1%) had higher mean scores compared with Cluster 1 for all items, and particularly for apparent sadness. Patients in Cluster 3 (18.9%) had the lowest mean scores for both apparent and reported sadness, but a similar profile compared with Cluster 1 for inner tension, reduced sleep, reduced appetite, and concentration difficulties. Cluster 4 (6.8%) had the highest mean scores for each item. Both apparent and reported sadness accounted for a large amount of variance among the four clusters. Patients in Cluster 4 were more likely to have 12 or less years of education and/or one or more functional limitations. The heterogeneity in symptom presentation among older adults diagnosed with major depression can potentially inform the development of DSM-V.


2,4-Dichlorophenoxyacetic Acid Poisoning Mimicking as Organophosphorus Poisoning.
Different compounds such as organochlorines, pyrethroids, fungicides, 2,4-dichlorophenoxy (2,4-D) herbicides, mushrooms, opioids, cartap compounds, and amitraz compounds can mimic organophosphorus (OP) poisoning. Muscle fasciculation, pulmonary edema, convulsions, bradycardia, hypotension, and smell caused by pyrethroids, as well as neurological signs, seizures, pulmonary edema, and smell caused by organochlorines can mimic OP poisoning. Miosis, vomiting, coma, and hypotension caused by opioids; miosis, bradycardia, altered sensorium, respiratory depression, and hypotension caused by amitraz compounds; and vomiting, breathlessness, altered sensorium, hypotension, and seizures caused by cartap compounds can also mimic OP poisoning. Mushroom poisoning and few fungicide compounds are also known to mimic features of OP poisoning. Hyperglycemia and glycosuria are the key hallmarks of amitraz poisoning. 2,4-D compounds can also mimic most of the features of OP poisoning; however, rhabdomyolysis, coma, and hyper/hypotonia are key differentiating features. Allergic manifestation and greenish discoloration of the contacted skin are the differentiating features of cartap poisoning. Treating all agriculture-related poisoning with atropine without confirming the compound can lead to a therapeutic misadventure. Here, we discuss the case of a patient who was referred to our Emergency Department (ED) with an alleged history of an unknown poison ingestion which was managed with atropinization for suspected OP poisoning in an outside hospital. On probing the history, the actual compound was found to be a 2,4-D herbicide. Very few documented case reports of 2,4-D poisoning are available in the literature. Hyper/hypotonia, coma, and skeletal muscle damage are the key differentiating features of 2,4-D poisoning. Our patient had skeletal muscle damage (rhabdomyolysis), evidenced by raised creatine kinase-total and creatine kinase-muscle/brain. As there is no specific antidote, we treated the patient with urinary alkalinization and supportive care. The patient had a favorable outcome in the ED.


Children's Microsystems and Their Relationship to Stress and Executive Functioning.
Microsystems are described as contexts formed by a subject, their roles, their interactions, and a specific physical space and time, such as housing and the school environment. Although several studies suggest the importance of studying this type of environment and its repercussion on children's development, in a Latin American context, few studies integrate the interaction of two primary settings in the development of executive functioning. The present study explores the effects of the quality of housing and school environments on the perception of stress, decision making, and planning among children. A total of 114 children (43% girls and 57% boys, M age = 10.57) from a primary school located in a community classified as poor participated in the study. The following was measured: the environmental quality of classrooms, housing, stress, and executive functioning of children. The results reveal a model linking environmental quality levels in children's homes and schools and executive functioning. We also obtain a mediating role of stress between microsystems and performance, finding a deficit in executive performance when children experienced higher levels of stress as a result of poor environmental quality both in their homes and in their schools.


An Italian-Australian comparison of quality of life among people with intellectual disability living in the community.
This paper reports the measurement of both objective and subjective quality of life in samples of people either with intellectual disability or from the general adult population, drawn from Australia and Italy. Measures were made using the Comprehensive Quality of Life Scale. Generally, it was found that the scores for all groups were comparable. This finding is consistent with the provision of generally decent objective living conditions in both countries and subjective well-being homeostasis, wherein subjective quality of life is held within a predictable range. These results are discussed in the context of measurement difficulties imposed by the need for simple Likert scales and subjective data which are strongly negatively skewed.


[Recovery of a complete cervical spinal cord injury with tetraplegia (author's transl)].
The prognosis of a complete cervical spinal cord injury is based on an accurate initial diagnosis and on immediate treatment in order to release the medullary compression and stabilise the spine. The diagnosis of a complete traumatic quadriplegia depends upon a thorough neurological examination, particularly noting sensation and motor power of the sacral segments, looking for a sacral sparing which is a sign of a better prognosis. The authors analysed a well documented case of progressive recovery obtained after nearly immediate open reduction and synthesis of a C1 C2 fracture-dislocation with complete quadriplegia.


A National Implementation Project to Prevent Healthcare-Associated Infections in Intensive Care Units: A Collaborative Initiative Using the Breakthrough Series Model.
Although there are simple and low-cost measures to prevent healthcare-associated infections (HAIs), they remain a major public health problem. Quality issues and a lack of knowledge about HAI control among healthcare professionals may contribute to this scenario. In this study, our aim is to present the implementation of a project to prevent HAIs in intensive care units (ICUs) using the quality improvement (QI) collaborative model Breakthrough Series (BTS). A QI report was conducted to assess the results of a national project in Brazil between January 2018 and February 2020. A 1-year preintervention analysis was conducted to determine the incidence density baseline of the 3 main HAIs: central line-associated bloodstream infections (CLABSIs), ventilation-associated pneumonia (VAP), and catheter-associated urinary tract infections (CA-UTIs). The BTS methodology was applied during the intervention period to coach and empower healthcare professionals providing evidence-based, structured, systematic, and auditable methodologies and QI tools to improve patients' care outcomes. A total of 116 ICUs were included in this study. The 3 HAIs showed a significant decrease of 43.5%, 52.1%, and 65.8% for CLABSI, VAP, and CA-UTI, respectively. A total of 5140 infections were prevented. Adherence to bundles inversely correlated with the HAI incidence densities: CLABSI insertion and maintenance bundle (R = -0.50, P = .010 and R = -0.85, P < .001, respectively), VAP prevention bundle (R = -0.69, P < .001), and CA-UTI insertion and maintenance bundle (R = -0.82, P < .001 and R = -0.54, P = .004, respectively). Descriptive data from the evaluation of this project show that the BTS methodology is a feasible and promising approach to preventing HAIs in critical care settings.


Label-free quantitative proteomic analysis of alfalfa in response to microRNA156 under high temperature.
Abiotic stress, including heat, is one of the major factors that affect alfalfa growth and forage yield. The small RNA, microRNA156 (miR156), regulates multiple traits in alfalfa during abiotic stress. The aim of this study was to explore the role of miR156 in regulating heat response in alfalfa at the protein level. In this study, we compared an empty vector control and miR156 overexpressing (miR156OE) alfalfa plants after exposing them to heat stress (40 °C) for 24 h. We measured physiological parameters of control and miR156OE plants under heat stress, and collected leaf samples for protein analysis. A higher proline and antioxidant contents were detected in miR156OE plants than in controls under heat stress. Protein samples were analyzed by label-free quantification proteomics. Across all samples, a total of 1878 protein groups were detected. Under heat stress, 45 protein groups in the empty vector plants were significantly altered (P < 0.05; |log2FC| > 2). Conversely, 105 protein groups were significantly altered when miR156OE alfalfa was subjected to heat stress, of which 91 were unique to miR156OE plants. The identified protein groups unique to miR156OE plants were related to diverse functions including metabolism, photosynthesis, stress-response and plant defenses. Furthermore, we identified transcription factors in miR156OE plants, which belonged to squamosa promoter binding-like protein, MYB, ethylene responsive factors, AP2 domain, ABA response element binding factor and bZIP families of transcription factors. These results suggest a positive role for miR156 in heat stress response in alfalfa. They reveal a miR156-regulated network of mechanisms at the protein level to modulate heat responses in alfalfa.


Method for localization of cloned DNA fragments on the Escherichia coli chromosome.
In exponentially growing cultures of Escherichia coli strains carrying the dnaC28 mutation, DNA replication can be synchronized by temperature changes (R. L. Rodriguez, M. S. Dalbey, and C. I. Davern, J. Mol. Biol. 74:599-604, 1973). We used this synchronization procedure and DNA-DNA hybridization to develop a technique for the localization of cloned chromosomal fragments on the genetic map. Because of the bidirectional nature of replication in E. coli, our method gave two possible positions (one on each replication arm). However because of the precision obtained for each position (+/- 1 map unit), the final mapping with various genetic techniques was greatly facilitated. Using this technique and a simple chromosomal mobilization test, we located at 93.2 +/- 1 min a cloned DNA fragment carrying an extragenic suppressor of dnaA46, a thermosensitive mutation in the dnaA initiation gene. Further analysis showed that the groES (mopA) and groEL (mopB) genes, both located at 94.2 min on the standard map, were indeed carried by the cloned suppressor fragment.


Descemet's Stripping-automated Endothelial Keratoplasty for symptomatic thioridazine deposits in the cornea.
The aim of this study was to use Descemet's Stripping-automated Endothelial Keratoplasty (DSAEK) as a novel treatment to remove thioridazine corneal deposits. A 53-year-old female presented with a 3-month history of visual loss and glare. She had been taking thioridazine (100 mg/day for 1 year) for a psychiatric disorder. Dense yellowish-brown deposits had developed in the posterior stroma. Thioridazine was discontinued, and she was switched to fluoxetine. One year after discontinuation of thioridazine, her symptoms and signs did not resolve. Standard DSAEK was performed on her left eye. Two weeks after DSAEK, an anterior subcapsular cataract was detected in the same eye. Phacoemulsification and intraocular lens implantation were performed after 3 months. The left cornea became completely clear after DSAEK, and the patient's best-corrected visual acuity improved from 20/40 to 20/20 at 1 month after cataract surgery. All-visual symptoms such as glare and halos improved postoperatively. We suggest that DSAEK can be used as a novel treatment to reduce vision problems caused by thioridazine-induced corneal deposits.


Molecular and immunohistochemical analyses of uveal melanoma patient cohort.
Uveal melanoma is a rare form of melanoma and the most frequent primary eye malignancy in adults. The major molecular alterations underlying uveal melanoma pathogenesis affect mainly the GNAQ, GNA11, SF3B1, and BAP1 genes. In this study, we somatically genotyped 31 Brazilian uveal melanomas for BRAF, GNA11, GNAQ, SF3B1, and BAP1 gene mutations and assessed BRCA2 and p53 protein expression. GNAQ and GNA11 mutations were detected in 60%, and SF3B1 mutation rate was 30%. p53 Immunostaining was markedly positive in 5/31, and 3/31 samples showed negative BRCA2 expression. This study supports the importance of these key genes in uveal melanoma tumorigenesis; p53 and BRCA pathways seem to play a role in a subset of patients, possibly heralding unfavorable prognosis.


Juvenile chronic myelogenous leukemia: characterization of the disease using cell cultures.
To characterize juvenile chronic myelogenous leukemia (JCML), the proliferative properties of bone marrow (BM) and peripheral blood (PB) cells from nine patients were studied using assays for CFU-C and CFU-GEMM and liquid cultures. All specimens showed two reproducible abnormalities: impaired growth of normal hematopoietic progenitors and excessive proliferation of monocyte-macrophage colonies in the absence of exogenous colony-stimulating activity (CSA). Cytogenetic studies in one patient indicated that the CFU-C were malignant because BM cells at diagnosis and monocyte-macrophage colonies showed an abnormal karyotype, whereas PB lymphocytes did not. In contrast to JCML, PB from six adults with Philadelphia (Ph1) chromosome-positive chronic myelogenous leukemia (Ph1 + CML) yielded CSA-dependent CFU-C colonies which were composed of granulocytes, macrophages, or both, as well as exuberant growth of BFU-E colonies. Co-cultures of JCML BM adherent or nonadherent cells with normal BM resulted in suppression of normal hematopoietic colony formation. Supernatant from JCML adherent cells in liquid culture or plasma from newly diagnosed untreated JCML patients also suppressed control BM colony growth in a dose-dependent manner. These findings confirm that JCML is a malignant disorder of monocytic lineage and suggest that the mechanism of hematopoietic failure in JCML is mediated by an inhibitory monokine secreted by malignant JCML cells.


A controlled clinical treatment trial of interpersonal psychotherapy for depressed pregnant women at 3 New York City sites.
While treatment decisions for antepartum depression must be personalized to each woman and her illness, guidelines from the American Psychiatric Association and the American College of Obstetrics and Gynecology include the recommendation of psychotherapy for mild-to-moderate depression in pregnant women. Although we previously demonstrated the efficacy of interpersonal psychotherapy for antepartum depression in a sample of Hispanic women, this study provides a larger, more diverse sample of African American, Hispanic, and white pregnant women from 3 New York City sites in order to provide greater generalizability. A 12-week bilingual, parallel-design, controlled clinical treatment trial compared interpersonal psychotherapy for antepartum depression to a parenting education program control group. An outpatient sample of 142 women who met DSM-IV criteria for major depressive disorder was randomly assigned to interpersonal psychotherapy or the parenting education program from September 2005 to May 2011. The 17-item Hamilton Depression Rating Scale (HDRS-17) was the primary outcome measure of mood. Other outcome scales included the Edinburgh Postnatal Depression Scale (EPDS) and the Clinical Global Impressions scale (CGI). The Maternal Fetal Attachment Scale (MFAS) assessed mother's interaction with the fetus. Although this study replicated previous findings that interpersonal psychotherapy is a beneficial treatment for antepartum depression, the parenting education program control condition showed equal benefit as measured by the HDRS-17, EPDS, CGI, and MFAS. This study supports the recommendation for the use of interpersonal psychotherapy for mild-to-moderate major depressive disorder in pregnancy. The parenting education program may be an alternative treatment that requires further study. ClinicalTrials.gov identifier: NCT00251043


MicroRNA-320 suppresses colorectal cancer by targeting SOX4, FOXM1, and FOXQ1.
Colorectal cancer (CRC) is the third most common cancer causing high mortality rates world-wide. Delineating the molecular mechanisms leading to CRC development and progression, including the role of microRNAs (miRNAs), are currently being unravelled at a rapid rate. Here, we report frequent downregulation of the microRNA miR-320 family in primary CRC tissues and cell lines. Lentiviral-mediated re-expression of miR-320c (representative member of the miR-320 family) inhibited HCT116 CRC growth and migration in vitro, sensitized CRC cells to 5-Fluorouracil (5-FU), and inhibited tumor formation in SCID mice. Global gene expression analysis in CRC cells over-expressing miR-320c, combined with in silico prediction identified 84 clinically-relevant potential gene targets for miR-320 in CRC. Using a series of biochemical assays and functional validation, SOX4, FOXM1, and FOXQ1 were validated as novel gene targets for the miR-320 family. Inverse correlation between the expression of miR-320 members with SOX4, FOXM1, and FOXQ1 was observed in primary CRC patients' specimens, suggesting that these genes are likely bona fide targets for the miR-320 family. Interestingly, interrogation of the expression levels of this gene panel (SOX4, FOXM1, and FOXQ1) in The Cancer Genome Atlas (TCGA) colorectal cancer data set (319 patients) revealed significantly poor disease-free survival in patients with elevated expression of this gene panel (P-Value: 0.0058). Collectively, our data revealed a novel role for the miR-320/SOX4/FOXM1/FOXQ1 axes in promoting CRC development and progression and suggest targeting those networks as potential therapeutic strategy for CRC.


Effects of Mg-ATP on the flavonoid production in the Scutellaria baicalensis Georgii suspension cultures.
This work focused on the cultivation of S. baicalensis Georgii in vitro cultures and on the possibilities of increasing the production of secondary metabolites in these cultures. The aim of the Sstudy was to determine whether the baicalin transport through vacuolar membrane is dependent on the presence of Mg-ATP. Our results showed that Mg-ATP had a significant effect on the ratio of baicalin and baicalein content and on the transport speed of these flavonoids. Therefore, the transport mechanism for baicalin are probably some of the MRP proteins which are the subfamily of the ABC transporte


The development of the effect of peer monitoring on generosity differs among elementary school-age boys and girls.
The purpose of this study was to examine the effect of peer monitoring on generosity in boys and girls aged 6-12 years. A total of 120 elementary school students played a one-shot dictator game (DG) with and without peer monitoring by classmates. Children decided how to divide 10 chocolates between themselves and a classmate either in a condition in which their allocations were visible to their peers, or in private. While the effect of peer monitoring on the allocation amount in the DG was clearly present in boys, it was not observed in girls. Furthermore, the effect of peer monitoring in boys appeared at the age of 9 years. These results suggest that the motivation to draw peers' attention plays a stronger role for older boys than for girls or younger boys. The potential roles of higher-order theory of mind, social roles, and emergence of secondary sex characteristics on the influence of peer monitoring on generosity shown by boys are discussed.


Amended structure of side chains in a cell wall mannan from Candida albicans serotype A strain grown in yeast extract-Sabouraud liquid medium under acidic conditions: detection of the branched side chains corresponding to antigenic factor 4.
In a previous study, we reported the excess production of alpha-1,3-linked mannose residues with the complete disappearance of beta-1,2-linked mannose residues in cell wall mannans of Candida albicans serotype A strain cells, which were grown in yeast extract-Sabouraud liquid medium at pH 2.0. In the present study, we examined the immunochemical reactivity of the same mannan of NIH A-207 with an enzyme-linked immunosorbent assay (ELISA) using several antisera to antigenic factors of the genus Candida (FAbs) and the structure of the mannan by two-dimensional homonuclear Hartmann-Hahn analysis. The ELISA showed that the mannan reacts to FAb 4 but not to FAbs 13b and 34, which are reported to be antibody factors against linear side chains containing an alpha-1,3-linked mannose residue. In the Hartmann-Hahn analysis, we found two branched side chains, Man alpha 1-2Man alpha 1-3[Man alpha 1-6]Man alpha 1-(2Man alpha 1-)(2)2Man and Man alpha 1-3[Man alpha 1-6]Man alpha 1-(2Man alpha 1-)(2)2Man, instead of the previously reported linear side chains. The branched side chains are oversynthesized under acidic conditions.


Influence of electrochemical polishing on the mechanical behaviour of nickel-titanium rotary files.
The purpose of this study was to evaluate the influence of electrochemical polishing on the cyclic fatigue, cutting efficiency and surface topography of Miltex nickel-titanium rotary files. A group of files were submitted to electrochemical polishing. New files and polished files were tested for flexural fatigue and for cutting efficiency. Scanning electron microscopy observations of all files were carried out to compare the surface morphologies of polished and new files. The t-test was used to compare the groups for cutting efficiency. Scanning electron microscopy analysis indicated that the polishing resulted in more regular surfaces in relation to the new files. Electropolished files attained a significantly higher number of cycles to fracture than new unpolished files. No statistical difference exists between the groups in relation to the cutting efficiency (t-test, P < 0.05). The polished files keep their integrity for a significantly higher number of cycles to fracture than the new files, without compromising cutting efficiency.


Philippines and Indonesia: on the way to a migration transition.
"This paper, in a comparative analysis of the Philippines and Indonesia, examines first under what conditions can migration favorably contribute to the process of economic development and then to what extent can economic growth impact upon reducing emigration pressures in these labor surplus economies. The paper also argues that there is still considerable scope for putting in place [an] agreed set of rules and policies to ensure better protection for the more vulnerable migrants."


Assessing the Risk of Omalizumab Add-on Therapy for Chronic Idiopathic Urticaria during the COVID-19 Pandemic.
Amid the current COVID-19 pandemic, there is concern for increased risk of infection while on immunomodulatory therapy. Omalizumab, a monoclonal antibody, is an add-on therapy for the treatment of chronic idiopathic urticaria (CIU) when first line therapy alone fails to achieve appropriate response. Current understanding of the response to COVID-19 infection is largely varied and actively under investigation. In the context of a pandemic, it is important to consider the safety profile of omalizumab as it modulates the immune system. We reviewed data from pivotal Phase III clinical trials investigating omalizumab use in CIU patients with a focus on reported respiratory-related adverse events (AEs) to assess these risks. Results from three phase III trials show that omalizumab adjunct therapy does not significantly increase infection risk and severity.


The dazzle reflex: electrophysiological signals from ocular muscles reveal strong binocular summation effects.
Under dark adapted or dim conditions the mammalian visual system is carefully programmed to respond rapidly to the sudden onset of bright lights. This response, called the dazzle reflex, is controlled from sub-cortical structures of the brain. It is known anecdotally that exposure to a bright light when dark adapted induces an instinctive closure of one eye to reduce the pain associated with dazzle. This binocular summation of the dazzle response has not previously been reported. The dazzle reflex can be measured in human subjects by recording the electrical activity from surface electrodes located near the muscles around the eye. In this paper we report an investigation of the apparent binocular summation of the dazzle reflex using this technique. The data reveal a clear difference between monocular and binocular stimulation, with the binocular response being much larger than the monocular response. Furthermore this monocular/binocular difference arises only if the stimulus duration is longer than approximately 1 s. These observations are interpreted in terms of the known physiology of blink mechanisms.


Crocetin: A Systematic Review.
Crocetin is an aglycone of crocin naturally occurring in saffron and produced in biological systems by hydrolysis of crocin as a bioactive metabolite. It is known to exist in several medicinal plants, the desiccative ripe fruit of the cape jasmine belonging to the Rubiaceae family, and stigmas of the saffron plant of the Iridaceae family. According to modern pharmacological investigations, crocetin possesses cardioprotective, hepatoprotective, neuroprotective, antidepressant, antiviral, anticancer, atherosclerotic, antidiabetic, and memory-enhancing properties. Although poor bioavailability hinders therapeutic applications, derivatization and formulation preparation technologies have broadened the application prospects for crocetin. To promote the research and development of crocetin, we summarized the distribution, preparation and production, total synthesis and derivatization technology, pharmacological activity, pharmacokinetics, drug safety, drug formulations, and preparation of crocetin.


Strain-induced differentiation of fetal type II epithelial cells is mediated via the integrin α6β1-ADAM17/tumor necrosis factor-α-converting enzyme (TACE) signaling pathway.
Mechanical forces are critical for normal fetal lung development. However, the mechanisms regulating this process are not well-characterized. We hypothesized that strain-induced release of HB-EGF and TGF-α is mediated via integrin-ADAM17/TACE interactions. Employing an in vitro system to simulate mechanical forces in fetal lung development, we showed that mechanical strain of fetal epithelial cells actives TACE, releases HB-EGF and TGF-α, and promotes differentiation. In contrast, in samples incubated with the TACE inhibitor IC-3 or in cells isolated from TACE knock-out mice, mechanical strain did not release ligands or promote cell differentiation, which were both rescued after transfection of ADAM17. Cell adhesion assay and co-immunoprecipitation experiments in wild-type and TACE knock-out cells using several TACE constructs demonstrated not only that integrins α6 and β1 bind to TACE via the disintegrin domain but also that mechanical strain enhances these interactions. Furthermore, force applied to these integrin receptors by magnetic beads activated TACE and shed HB-EGF and TGF-α. The contribution of integrins α6 and β1 to differentiation of fetal epithelial cells by strain was demonstrated by blocking their binding site with specific antibodies and by culturing the cells on membranes coated with anti-integrin α6 and β1 antibodies. In conclusion, mechanical strain releases HB-EGF and TGF-α and promotes fetal type II cell differentiation via α6β1 integrin-ADAM17/TACE signaling pathway. These investigations provide novel mechanistic information on how mechanical forces promote fetal lung development and specifically differentiation of epithelial cells. This information could be also relevant to other tissues exposed to mechanical forces.


Menstrual cycle alters sympathetic neural responses to orthostatic stress in young, eumenorrheic women.
Sympathetic baroreflex sensitivity (BRS) and muscle sympathetic nerve activity (MSNA) responses during early follicular (EF) and midluteal (ML) phases of the menstrual cycle are controversial. We hypothesize an augmented sympathetic BRS and MSNA response to orthostatic stress during the ML phase of the menstrual cycle. MSNA, mean arterial pressure (MAP), and heart rate (HR) were recorded during progressive lower body negative pressure (LBNP) (-5, -10, -15, -20, -30, and -40 mmHg; 3 min/stage) in 13 healthy, eumenorrheic women (age 21 +/- 1 yr). Sympathetic BRS was assessed by examining relations between spontaneous fluctuations of diastolic arterial pressure and MSNA at rest and during progressive LBNP. Plasma estradiol (42 +/- 6 vs. 112 +/- 12 pg/ml; P < 0.01) and progesterone (2 +/- 0 vs. 10 +/- 2 ng/ml; P < 0.04) were elevated during the ML phase. Resting MSNA (8 +/- 1 vs. 11 +/- 1 bursts/min), MAP (79 +/- 2 vs. 78 +/- 2 mmHg), and HR (58 +/- 2 vs. 60 +/- 2 beats/min) were not different during EF and ML phases. MSNA and HR increased during progressive LBNP (P < 0.001), and the increases in MSNA burst frequency (bursts/min) and HR were similar during both phases. In contrast, increases in total MSNA (arbitrary units) during progressive LBNP were augmented during the ML phase (P < 0.04), but this response does not appear to be linked to differences in sympathetic BRS. Progressive LBNP did not change MAP during either phase. Our results demonstrate an augmentation of the MSNA response to progressive LBNP during the ML phase of the menstrual cycle. These findings suggest that hormonal fluctuations of eumenorrheic women may influence sympathoexcitation during an orthostatic challenge, but not through sympathetic baroreflex-mediated pathways.


K-band high-pressure cell for ENDOR studies.
This article describes a versatile high-pressure K-band EPR-ENDOR system, designed for the examination of the effects of hydrostatic pressure on F-center hyperfine interactions. Dielectric filling is used to minimize the inner dimensions of the pressure cell. Cavity mode and coil geometry permit the same system to be used either for high-frequency field modulation EPR or for ENDOR by rotating the magnetic field. The pressure seal is also part of a rotatable microwave joint permitting the microwave coupling to be varied by rotating the cavity-pressure cell assembly. The beryllium-copper high-pressure vessel is separated from the pressure generating device by high-pressure transmitting tubing. It has been tested to 18 kilobars.


Differential Histone Posttranslational Modifications Induced by DNA Hypomethylating Agents.
The prototype DNA hypomethylating agents 5-azacytidine (5AC) and decitabine (DAC) are currently FDA-approved for treatment of blood and bone marrow disorders like myelodysplastic syndrome. 5AC and DAC are considered similar drugs and were shown to induce histone modifications that modulate gene expression. The aim of this study is to compare the effect of both drugs on histone acetylation and methylation at multiple histone amino acids residues. Mass spectrometry was used to compare the effect of both drugs on 95 different histone posttranslational modifications (PTMs) in leukemia cells. ChIP-Seq analysis was used to compare the impact of both drugs on the genome-wide acetylation of the H3K9 mark using primary leukemia cells from six de-identified AML patients. Both DAC and 5AC induced histone PTMs in different histone isoforms like H1.4, H2A, H3, H3.1, and H4. Changes in both histone methylation and acetylation were observed with both drugs; however, there were distinct differences in the histone modifications induced by the two drugs. Since both drugs were shown to increase the activity of the HDAC SIRT6 previously, we tested the effect of 5AC on the acetylation of H3K9, the physiological substrate SIRT6, using ChIP-Seq analysis and compared it to the previously published DAC-induced changes. Significant H3K9 acetylation changes (P< .05) were detected at 925 genes after 5AC treatment vs only 182 genes after DAC treatment. Nevertheless, the gene set modified by 5AC was different from that modified by DAC with only ten similar genes modulated by both drugs. Despite similarity in chemical structure and DNA hypomethylating activity, 5AC and DAC induced widely different histone PTMs and considering them interchangeable should be carefully evaluated. The mechanism of these histone PTM changes is not clear and may involve modulation of the activity or the expression of the enzymes inducing histone PTMs.


Isolation and nitrogen-fixing activity of Frankia sp. strain CpI1 vesicles.
Under N2-fixing conditions in aerobic culture and in symbiosis, frankiae produce spherical, multicellular structures that have been called vesicles. The vesicles have been proposed as the site of nitrogen fixation. We isolated vesicles by using density centrifugation in a single-step sucrose gradient. Vesicles migrated out of 50% (wt/vol) sucrose and banded at the 40 to 50% sucrose interface; they were intact, as assessed by transmission electron microscopy, and were free of hyphal contamination. Specific activities of nitrogenase in vesicles prepared anaerobically were up to 100-fold greater than the specific activity of the largely hyphal pellet, depending on the recovery of vesicles. All of the activity in the pellet could be accounted for by the number of vesicles present in the pellet. Glutamine synthetase activity in crude extracts of vesicles was extremely low.


Potentiation of 5-HT3 receptor-mediated responses by protein kinase C activation.
The effect of agents that activate or inhibit protein kinase C (PKC) on the function of recombinant 5-HT3 receptors expressed in Xenopus oocytes was studied. The PKC activator phorbol 12-myristate 13-acetate (PMA) induced a long-lasting increase in the amplitude of 5-HT-activated ion current. The potentiation was maximal at 20 min and had a duration of approximately 60 min. The inactive phorbol ester, 4 alpha-PMA, had no effect on 5-HT3 receptor-mediated current. The PMA-induced potentiation was concentration-dependent over the concentration range 0.1-300 nM. The percentage potentiation by PMA was maximal at low 5-HT concentrations and decreased with increasing concentrations of 5-HT. For current activated by 0.1 microM 5-HT, maximal potentiation (Emax) was 667% of control, the EC50 was 15 nM and the apparent Hill coefficient was 0.99. The PKC inhibitor, staurosporin, antagonized the PMA potentiation; whereas, inhibitors of protein kinase A (PKA) or tyrosine kinase had no effect on this potentiation. The observations show that PMA can potentiate 5-HT3 receptor-mediated responses and suggest that this potentiation is mediated by activation of PKC.


First-degree relatives of early-onset gastric cancer patients show a high risk for gastric cancer: phenotype and genotype profile.
First-degree relatives (FDR) of early-onset gastric cancer (EOGC) is presumed to be a population with a distinct molecular and phenotypic profile, regarding the prevalence of gastric premalignant conditions and the association with Helicobacter pylori infection and host proinflammatory gene polymorphisms. A case-control study was conducted with FDR of EOGC patients (n = 103) and age and gender matched controls (n = 101; ranging from spouses to neighbors and dyspeptics). Upper endoscopy was performed, Operative Link on Gastritis Assessment (OLGA) system used for staging and H. pylori (cagA and vacA) and host IL1B-511, IL1RN intron2 VNTR and IFNGR1-56 genotyping. Seventy percent of cases showed atrophy, while 19 % presented with high-stage gastritis (OLGA stage III or IV) (p < 0.001); gastric dysplasia was present in seven cases (vs none in controls) (p = 0.007). In cases, H. pylori was present in 82 % (vs 62 % in controls; p = 0.004) with vacA s1 and vacA m1 + strains significantly associated with the presence of atrophy; individuals homozygous for IL1B-511*T present a significantly higher risk for dysplasia. An increased global prevalence of IFNGR1-56*T/*T polymorphism (37 % in cases vs 24 % in controls; p = 0.03) was observed with no association with atrophic changes or dysplasia. All trends observed were kept when comparing FDR of EOGC with spouses, neighbors, or dyspeptic controls. We demonstrated that FDR of EOGC patients have an increased prevalence of high-risk OLGA stages and dysplasia that seem to be associated with high virulence H. pylori strains and pro-inflammatory host genotypes, including a possible population-specific risk marker. FDR of EOGC patients may merit specific management through endoscopic and histopathological adequate assessment of gastric mucosa and surveillance.


NMR solution structure and dynamics of the peptidyl-prolyl cis-trans isomerase domain of the trigger factor from Mycoplasma genitalium compared to FK506-binding protein.
We have solved the solution structure of the peptidyl-prolyl cis-trans isomerase (PPIase) domain of the trigger factor from Mycoplasma genitalium by homo- and heteronuclear NMR spectroscopy. Our results lead to a well-defined structure with a backbone rmsd of 0.23 A. As predicted, the PPIase domain of the trigger factor adopts the FK506 binding protein (FKBP) fold. Furthermore, our NMR relaxation data indicate that the dynamic behavior of the trigger factor PPIase domain and of FKBP are similar. Structural variations when compared to FKBP exist in the flap region and within the bulges of strand 5 of the beta sheet. Although the active-site crevice is similar to that of FKBP, subtle steric variations in this region can explain why FK506 does not bind to the trigger factor. Sequence variability (27% identity) between trigger factor and FKBP results in significant differences in surface charge distribution and the absence of the first strand of the central beta sheet. Our data indicate, however, that this strand may be partially structured as "nascent" beta strand. This makes the trigger factor PPIase domain the most minimal representative of the FKBP like protein family of PPIases.


[Effects of intercropping Chinese onion cultivars of different allelopathic potential on cucumber growth and soil micro-environment].
A pot experiment was conducted to study the effects of intercropping various Chinese onion cultivars of different allelopathic potential on the cucumber growth and rhizospheric soil environment. When intercropped with high allelopathic Chinese onion cultivars, the EC value and peroxidase activity of cucumber rhizospheric soil decreased, while the pH value, invertase and catalase activities, and bacterial community diversity increased. The cloning and sequencing results indicated that most DGGE bands amplified from cucumber rhizospheric soil samples showed a high homology to uncultured bacterial species. The common bands were affiliated with Actinobacteria and Proteobacteria, and the differential bacteria bands were affiliated with Proteobacteria and Anaerolineaceae. Rhodospirillales and Acidobacteria were only found in the cucumber rhizospheric soil intercropped with low allelopathic Chinese onion cultivars. Correlation analysis showed that there were significant positive correlations between rhizospheric soil urease activity and cucumber seedlings height, total dry biomass, leaf area, and DGGE band number. It was suggested that intercropping high allelopathic Chinese onion cultivars could establish a good rhizospheric soil micro-environment for cucumber growth, and promote the growth of cucumber seedlings markedly.


Light trapping efficiency of organic solar cells with large period photonic crystals.
We study the optical properties of a 2D Photonic Crystal (PC) inserted in the upper ITO electrode of a classical P3HT:PCBM solar architecture with an ultra-thin active layer. First, we analyze the optical response of the system when only the active layer is supposed to absorb light. This allows us to observe clear photonic crystal resonances in the absorption spectrum, which increase the cell efficiency even if the period of the PC is higher than the wavelength. This is in apparent contradiction with the common belief that PC should work in subwavelength regime. Then, by turning to a real system (with optical losses in all the layers), an optimized PC design is proposed, where the maximum of efficiency is obtained for a PC period of 1200 nm, much larger than visible wavelength.


Construction and characterization of mutant iso-2-cytochromes c with replacement of conserved prolines.
Oligonucleotide-directed mutagenesis has been used to construct two mutant forms of iso-2-cytochrome c. In one, Pro-30 is replaced by threonine; in the other, Pro-76 is replaced by glycine. Both prolines are fully conserved among mitochondrial cytochromes c and play important structural and functional roles. Yeast with either the Pro-30 or the Gly-76 mutation has appreciable levels of mutant protein in vivo and grows on media containing nonfermentable carbon sources. Thus, neither mutation blocks protein targeting to mitochondria, uptake by mitochondria, covalent attachment of heme, or in vivo function. As judged by ultraviolet-visible spectrophotometry and proton nuclear magnetic resonance spectroscopy, the nativelike conformation of purified Gly-76 iso-2 at pH 6 is almost indistinguishable from that of the normal protein at pH 6. Ultraviolet second-derivative spectrophotometry, however, suggests an increase in the average number of exposed tyrosine side chains, with 2.25 out of 5 residues exposed for the mutant compared to 1.95 for normal iso-2. Above neutral pH, the protein folds to a mutant conformation possibly related to alkaline cytochrome c. Nuclear Overhauser difference spectroscopy of the reduced nativelike conformation allows assignment of several proton resonances and comparison of side-chain conformations of the heme ligand Met-80 in the mutant and the normal proteins. The proton chemical shifts for the assigned resonances are the same within errors for Gly-76 iso-2 and normal iso-2 at pD 6, 20 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)


Cost-effectiveness of nonpharmacologic, nonsurgical interventions for hip and/or knee osteoarthritis: systematic review.
To investigate the cost-effectiveness of nonpharmacological, nonsurgical interventions for the treatment of hip and/or knee osteoarthritis. We identified economic evaluations or cost studies associated with randomized or quasi-randomized controlled trials that assessed nonpharmacologic, nonsurgical interventions for the treatment of hip and/or knee osteoarthritis. Medline, Embase, PubMed, National Health Service Economic Evaluation Database, CENTRAL, EconLit, and OpenSIGLE were searched up to October 1, 2010. Study characteristics extracted include study population, health outcomes, and economic analysis elements. Economic analyses were assessed by using the Quality of Health Economic Studies instrument, and the methodological quality of the randomized controlled trials was graded by using an internal validity checklist. All costs were converted to 2008 US dollars. Ten economic evaluations and one randomized controlled trial reporting health-care costs met our inclusion criteria. Interventions included exercise programs, acupuncture, rehabilitation programs, and lifestyle interventions. Six of the 11 studies exhibited high risks of bias for the cost and/or effect components of their cost-effectiveness estimate. Six studies used comparators of unknown cost-effectiveness. Four studies reported cost-effectiveness estimates lower than $50,000 per quality-adjusted life-year. All studies evaluating exercise interventions found the programs to be cost saving. There is only limited evidence for the cost-effectiveness of conservative treatments for the management of hip and/or knee osteoarthritis. More high-quality economic evaluations of conservative interventions are needed to further inform practice.


[Smoking and alimentary diseases].
Tobacco smoking directly or hematogenously acts on digestive organ and induces benign and malignant alimentary diseases. Nicotine, nitrosamines, and polycyclic aromatic hydrocarbons included in tobacco smoke are major causative agents of tobacco smoking-related alimentary diseases. These agents have harmful effects on digestive organ through compromised blood circulation, impaired neural regulation, and cell dysfunction. These malfunctions cause tissue destruction, such as chronic inflammation and ulcer formation. Besides forming DNA adducts, they also act on endogenous signal transduction, leading to gene mutation and abnormal cell death and growth. Interestingly, in inflammatory diseases, smoking improves symptoms of ulcerative colitis, while it worsens the condition of Crohn diseases. Smoking is also involved in lately increasing Barrette's esophagus. Future studies are needed.


Abdominal regional fat distribution on MRI correlates with cholecystolithiasis.
To determine whether abdominal regional fat distribution pattern on MRI is correlated with cholecystolithiasis. Magnetic resonance imaging (MRI) of 163 patients with cholecystolithiasis and 163 non-cholecystolithiasis control subjects admitted to our institution between March 2011 and September 2013 were included in this cross-sectional evaluation. There were 98 women and 65 men in cholecystolithiasis group with an average age of 57±16 years (range 25-86 years). There were 87 women and 76 men in the control group with an average age of 41±16 years (range 14-77 years). Visceral adipose tissue (VAT), abdominal subcutaneous adipose tissue (SAT) and total abdominal adipose tissue (TAT) of all the subjects at navel level were measured on abdominal MRI. According to the visceral adipose area (cut-off point VAT = 100 cm2), study subjects were divided into 1) increased accumulation of intra-abdominal fat and 2) normal distribution of intra-abdominal fat. Logistic regression was used to assess the association of fat with the presence of cholecystolithiasis, adjusted for age and sex. The incidence of increased intra-abdominal fat accumulation in the cholecystolithiasis group was significantly higher than that of the control group (P = 0.000). After adjusting for age and sex, cholecystolithiasis was associated with a one standard deviation increment in the waist circumference (WC) (OR = 1.44; 95%CI: 1.01,1.93; p = 0.00), VAT (OR = 4.26; 95%CI: 1.85,5.29; p = 0.00), VAT/SAT (OR = 8.66; 95%CI: 1.60,12.63; p = 0.00), and VAT/TAT (OR = 6.73; 95%CI: 4.24,12.18; p = 0.00), but not with fat content in the abdominal subcutaneous fat (p = 0.19). The visceral adipose tissue and distribution proportion of abdominal adipose tissue are correlates of cholecystolithiasis.


Introduction of the American/Asian genotype of dengue 2 virus into the Yucatan State of Mexico.
A dengue (DEN) outbreak occurred in the Yucatan State of Mexico in 2002. Three isolates were obtained from patients presenting with DEN-like symptoms, and examined by partial nucleotide sequencing and phylogenetic analysis. The isolates were identified as DEN-2 viruses of the American-Asian genotype; this is the first report of this genotype in the Yucatan State. The DEN-2 viruses of the American-Asian genotype have been associated with more severe disease outcomes. Thus, its introduction into the Yucatan State presents a serious problem to public health authorities. During this outbreak, DEN virus infection was confirmed in 18% (282 of 1,560) of the patients who presented with DEN-like symptoms. Of these, 87 (31%) patients met the World Health Organization criteria for dengue hemorrhagic fever, including two patients who died. The majority (77%) of the patients experienced secondary infections in this epidemic.


[Evaluation of the Immulite 2000 Toxoplasma quantitative IgG et Toxoplasma IgM for the diagnosis of human toxoplasmosis].
Four hundred and ninety five human sera with clinical and biological data were tested for the evaluation of Immulite 2000 Toxoplasma Quantitative IgG and Immulite 2000 Toxoplasma IgM produced by Diagnostic Products Corporation (Los Angeles, USA) for the diagnosis of human toxoplasmosis. The results of these kits were compared to those of the University Hospital of Nancy where the reference assays were Enzygnost Toxoplasmosis IgG and Enzygnost Toxoplasmosis IgM (Berhing-Dade, Germany), Toxoscreen (bioMérieux, France), ISAgA Plus (IgM et IgA) (bioMérieux, France). The sensitivity and the specificity of IgG detection by Immulite 2000 Toxoplasma Quantitative IgG were 98% and 100%, respectively. The high sensitivity of IgM detection by Immulite 2000 Toxoplasma IgM was adapted to the early diagnosis of toxoplasmic primo-infection and to the pediatric diagnosis or follow-up of congenital toxoplasmosis but could reveal IgM a long time after primary infection.


[Hemorrhoidal disease and the concomitant changes in the rectosigmoid].
Hemorrhoids is the most common disease in coloproctology, and rectoromanoscopy is a valuable tool of diagnosis. Of a total of 1614 patients examined at the proctologic consulting room of the Department of Gastroenterology at the Medical Academy in Sofia, hemorrhoids had 609 (37.8 per cent). External were 2 per cent of the hemorrhoids, internal 81.1 per cent and external + internal 15.9 per cent. Characteristic finding in internal hemorrhoids were the swelling in the anal canal, localized along 3, 7 and 11 hours of the clock dial in knee-elbow position of the patient. Endoscopic symptoms of hemorrhoid malignization were ulceration, thickening and infiltration of the mucosa at the bottom of the erosion. Hemorrhoids were frequently associated with accompanying diseases (52.6 per cent) which aggravated the patients complaints. They may occasionally be the cause of symptomatic hemorrhoids.


DNA Adsorption Studies of Poly(4,4'-Cychlohexylidene Bisphenol Oxalate)/Silica Nanocomposites.
The present study deals with the synthesis, characterization, and DNA extraction of poly(4,4'-cyclohexylidene bisphenol oxalate)/silica (Si) nanocomposites (NCs). The effects of varying the monomer/Si (3.7%, 7%, and 13%) ratio towards the size and morphology of the resulting NC and its DNA extraction capabilities have also been studied. For the NC synthesis, two different methods were followed, including the direct mixing of poly(4,4'-cyclohexylidene bisphenol oxalate) with fumed Si, and in situ polymerization of the 4,4'-cyclohexylidene bisphenol monomer in the presence of fumed silica (11 nm). The formed NCs were thoroughly investigated by using different techniques such as scanning electron microscopy (SEM), fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), powdered X-ray diffraction (XRD), and Brunauer-Emmett-Teller (BET) analysis where the results supported that there was the successful formation of poly(4,4'-cyclohexylidene bisphenol oxalate)/Si NC. Within the three different NC samples, the one with 13% Si was found to maintain a very high surface area of 12.237 m²/g, as compared to the other two samples consisting of 7% Si (3.362 m²/g) and 3.7% Si (1.788 m²/g). Further, the solid phase DNA extraction studies indicated that the efficiency is strongly influenced by the amount of polymer (0.2 g > 0.1 g > 0.02 g) and the type of binding buffer. Among the three binding buffers tested, the guanidine hydrochloride/EtOH buffer produced the most satisfactory results in terms of yield (1,348,000 ng) and extraction efficiency (3370 ng/mL) as compared to the other two buffers of NaCl (2 M) and phosphate buffered silane. Based on our results, it can be indicated that the developed poly(4,4'-cyclohexylidene bisphenol oxalate)/Si NC can serve as one of the suitable candidates for the extraction of DNA in high amounts as compared to other traditional solid phase approaches.


In vivo imaging of tagged mRNA in plant tissues using the bacterial transcriptional antiterminator BglG.
RNA transport and localization represent important post-transcriptional mechanisms to determine the subcellular localization of protein synthesis. Plants have the capacity to transport messenger (m)RNA molecules beyond the cell boundaries through plasmodesmata and over long distances in the phloem. RNA viruses exploit these transport pathways to disseminate their infections and represent important model systems to investigate RNA transport in plants. Here, we present an in vivo plant RNA-labeling system based on the Escherichia coli RNA-binding protein BglG. Using the detection of RNA in mobile RNA particles formed by viral movement protein (MP) as a model, we demonstrate the efficiency and specificity of mRNA detection by the BglG system as compared with MS2 and λN systems. Our observations show that MP mRNA is specifically associated with MP in mobile MP particles but hardly with MP localized at plasmodesmata. MP mRNA is clearly absent from MP accumulating along microtubules. We show that the in vivo BglG labeling of the MP particles depends on the presence of the BglG-binding stem-loop aptamers within the MP mRNA and that the aptamers enhance the coprecipitation of BglG by MP, thus demonstrating the presence of an MP:MP mRNA complex. The BglG system also allowed us to monitor the cell-to-cell transport of the MP mRNA, thus linking the observation of mobile MP mRNA granules with intercellular MP mRNA transport. Given its specificity demonstrated here, the BglG system may be widely applicable for studying mRNA transport and localization in plants.


Diuretic Action of Sodium-Glucose Cotransporter 2 Inhibitors and Its Importance in the Management of Heart Failure.
Primarily, the sodium-glucose cotransporter 2 (SGLT2) inhibitors suppress the cotransport of glucose and sodium from the tubular lumen of the proximal tubules to the blood, and excrete glucose into the urine. Therefore, glucose and caloric balances become negative, reducing both the blood glucose level and insulin secretion. On the other hand, the proximal tubular fluid, constituted with low chloride concentration because of SGLT2 inhibition, is transferred to the loop of Henle. Under low chloride conditions, the reabsorption mechanisms in the loop of Henle do not work, similar to when loop diuretics are given. Subanalysis data on heart failure (HF) from the EMPA-REG OUTCOME trials are discussed, assuming that SGLT2 inhibitors are loop diuretics. Renin-angiotensin system inhibitors and β-blockers contribute to prognostic improvements of HF, independent of SGLT2 inhibitors, and therefore, both regimens are essential for the treatment of HF. On the other hand, the prognostic improvements by SGLT2 inhibitors are not significant under treatment including conventional diuretics such as loop diuretics and aldosterone antagonists, suggesting that the prognostic improvement in HF by SGLT2 inhibitors is mostly through their diuretic action. (Circ J 2016; 80: 2277-2281).


Phase I clinical trial of decitabine (5-aza-2'-deoxycytidine) administered by hepatic arterial infusion in patients with unresectable liver-predominant metastases.
DNA demethylating agents may increase the immunogenicity of malignant tumours and increase the efficacy of subsequent treatment with immune check point inhibitors. We investigated the safety of administrating the demethylating agent decitabine by hepatic arterial infusionin patients with unresectable liver meta stases from solid tumours in a dose escalation phase I clinical trial. A total of nine eligible patients were enrolled and initiated study treatment at three different dose levels (two patients at 10, four at 15 and six at a dose level of 20mg decitabine/m2/day) (per protocol there was no intent to escalate the dose above the median tolerated intravenous dose level). Decitabine was administered as a 1-hour hepatic arterial infusion on five consecutive days every 4 weeks. Intrapatient dose escalation was applied in five patients. Grades 1 and 2 haematological toxicity was the most frequent treatment-related adverse event. None of the patients experienced treatment-limiting adverse events. Expression analysis of 30 cancer test is antigens (CTA) in pretreatment and post-treatment biopsies from patients indicated an increased expression of 21 CTAs after treatment. There were no objective tumour responses on study treatment or during post study exposure to immune checkpoint therapy in four patients with uveal melanoma liver metastases. We conclude that the investigate d hepatic arterial administration regimen for decitabine can be safely applied, and a dose level of 20 mg/m2/day on five consecutive days every 4 weeks can be considered for further investigation in combinatorial immunotherapy regimens. NCT02316028.


Lipoid pneumonia in 53 patients after aspiration of mineral oil: comparison of high-resolution computed tomography findings in adults and children.
To evaluate the high-resolution computed tomography (HRCT) findings in 53 patients with exogenous lipoid pneumonia and to compare the imaging features of adults and children. The study included 35 children and 18 adults. Statistical comparisons of findings in the 2 age groups were performed using either Pearson chi2 or Fisher exact test, as appropriate, at 5% significance level. The main HRCT findings included air-space consolidation, ground glass attenuation, air-space nodules, and crazy-paving pattern. Abnormalities predominated in the posterior regions of the right lung. The right lower lobe was most likely to show severe involvement. The presence of air-space consolidation, the involvement of upper right lobe, and the central and posterior distribution of the lesions were more common in children, whereas the crazy-paving pattern and random localization were significantly more frequent in adults. The other findings were not different between the 2 groups.


Isolation and identification of a protein binding to the localization element of Metallothionein-1 mRNA.
mRNA localization provides a mechanism for localized protein synthesis. mRNAs encoding certain proteins, including c-MYC, c-FOS, MT-1 (Metallothionein-1) and vimentin, are localized around the nuclei of mammalian cells and are associated with the cytoskeleton. Targeting of these mRNAs to the perinuclear cytoplasm is mediated by elements within their 3'-UTRs (3'-untranslated regions), but many of the trans-acting proteins remain unidentified. UV cross-linking assays using radiolabelled transcripts indicated that a protein of approx. 50 kDa (from the Chinese-hamster ovary cell extracts) bound to the MT-1 3'-UTR sequence. Competition experiments using unlabelled mutant 3'-UTR RNAs revealed that the binding of this protein is specific to localization-positive mutants. Isolation of a 50 kDa protein was achieved by an RNA affinity-based method in which biotinylated MT-1 3'-UTR RNA was anchored to paramagnetic beads. Bound proteins were eluted and analysed by SDS/PAGE. The 50 kDa protein was extracted from the gel, subjected to trypsin digestion and identified by matrix-assisted laser-desorption/ionization-time-of-flight mass spectrometry as eukaryote elongation factor 1alpha.


Recurrent Indoor Environmental Pollution and Its Impact on Health and Oxidative Stress of the Textile Workers in Bangladesh.
Perennial indoor environmental pollution in the textile industrial area is a potential health hazard for workers engaged in this line of work, resulting in mental aberration to severe health risks. This study was designed to investigate the indoor environmental quality of textile industries and correlate its effect on the occupational health and well-being of the textile workers by measuring plasma oxidative stress status in textile workers and healthy control subjects. Environmental samples were collected from 15 textile industries located in Dhaka division, and 30 volunteer textile workers and 30 volunteer office workers (control) aged 18 to 57 years participated in the study. The concentration of plasma ascorbic acid (P-ASC), plasma malondialdehyde (P-MDA), and plasma conjugated diene (P-CD) was measured in both groups. The noise level (78.0 ± 0.68 dB) and the formaldehyde level (141.80 ± 4.47 µg/m3) were found to be significantly higher in the indoor environmental area compared with those in the control area (70.17 ± 0.25 dB and 108.0 ± 0.76 µg/m3, respectively). Furthermore, the daily average concentration of suspended particulate matters (PMs), that is, PM2.5 (322.2 ± 13.46 µg/m3) and PM10 (411.0 ± 17.57 µg/m3), was also found to be significantly higher in the indoor environmental air compared with that in the control area (78.59 ± 1.66 and 174.0 ± 2.33 µg/m3, respectively). The levels of P-MDA (0.37 ± 0.03 nmol/L) and P-CD (14.74 ± 0.61 nmol/L) were significantly increased, whereas the level of P-ASC level (0.46 ± 0.04 mg/dL) was markedly decreased in the textile workers compared with the healthy control subjects (0.18 ± 0.01 nmol/L of P-MDA, 10.04 ± 0.44 nmol/L of P-CD, and 1.29 ± 0.06 mg/dL of P-ASC). The textile plants were found to have significantly elevated levels of indoor environmental pollutants compared with those in the control area, and the textile workers were significantly exposed to oxidative stresses compared with the control subjects. The use of noise pads and high-efficiency air filters is perhaps highly instrumental to put an end to this prevailing situation. Moreover, to overcome the oxidative stresses among workers, supplementation of antioxidant vitamins (ie, ascorbic acid and/or vitamin E) may be beneficial. In addition, to prevent serious health-related issues, proper precautions should be taken to protect the occupational health of the textile workers.


High Abundances of Microplastic Pollution in Deep-Sea Sediments: Evidence from Antarctica and the Southern Ocean.
Plastic pollution in Antarctica and the Southern Ocean has been recorded in scientific literature since the 1980s; however, the presence of microplastic particles (<5 mm) is less understood. Here, we aimed to determine whether microplastic accumulation would vary among Antarctic and Southern Ocean regions through studying 30 deep-sea sediment cores. Additionally, we aimed to highlight whether microplastic accumulation was related to sample depth or the sediment characteristics within each core. Sediment cores were digested and separated using a high-density sodium polytungstate solution (SPT) and microplastic particles were identified using micro-Fourier-transform infrared spectroscopy (μFTIR). Microplastic pollution was found in 93% of the sediment cores (28/30). The mean (±SE) microplastics per gram of sediment was 1.30 ± 0.51, 1.09 ± 0.22, and 1.04 ± 0.39 MP/g, for the Antarctic Peninsula, South Sandwich Islands, and South Georgia, respectively. Microplastic fragment accumulation correlated significantly with the percentage of clay within cores, suggesting that microplastics have similar dispersion behavior to low density sediments. Although no difference in microplastic abundance was found among regions, the values were much higher in comparison to less remote ecosystems, suggesting that the Antarctic and Southern Ocean deep-sea accumulates higher numbers of microplastic pollution than previously expected.


[Percutaneous endoscopic gastrostomy (PEG)].
Percutaneous endoscopic gastrostomy (PEG), although a standard method for long-time enteral nutrition, still leaves uncertainty about details in indication, technique and complications. Based on own experiences (approximately 1000 PEG's and 170 EPJ's in the last 10 years) these topics are discussed. As new indications the PEG for decompression in chronic intestinal obstruction and the questionable relative contraindications ascites, peritoneal carcinosis and Morbus Crohn are discussed. The common thread-pull through-technique is recommended in a slightly modified variation, according to direct puncture techniques of small intestine: the cannulas inserted in the intestine are fixed by an endoscopic alligator forceps, thus facilitating precise puncture and raising safety for the patient due to avoidance of dislocation. As complications local infections, implantation metastases and the buried bumper syndrome have to be mentioned and avoided. Alternative methods for PEG and different exchange systems for more comfort for the patients are discussed finally.


No inhibition of interferon gamma release in human lymphocytes by Ciamexone.
In previous experiments Ciamexone, derivative of 2-cyan-aziridine, was able to influence T-cell-mediated regulatory mechanisms but seemed to have no or only little effect on T cell effector mechanisms. On the basis of these observations Ciamexone seems to be a highly selective immunosuppressive agent. In order to evaluate further possible mechanisms of Ciamexone it was the aim of our investigation to study its influence on interferon gamma (IFN gamma) production in phytohaemogglutinin (PHA)-stimulated T lymphocytes of 15 tumour patients and 12 healthy reference subjects. The IFN gamma concentration of the cell supernatant was measured using an enzyme-linked immunosorbent assay. When the cells were stimulated with PHA at 7.5 micrograms/ml the IFN gamma concentration rose to significantly different values in the reference group (1.0 ng/ml) as compared to the tumour patients (0.4 ng/ml) (P less than 0.05). An addition of Ciamexone (at any of the concentrations administered) to PHA stimulation of peripheral blood mononuclear cells (PBMC) showed no influence on the IFN gamma release in either test group. The influence of hydrocortisone on the stimulation of PBMC with PHA resulted in a dose-dependent suppression of IFN gamma production in both test groups, again with significant differences between them. The IFN gamma concentration was 0.95 ng/ml in the reference group and 0.2 ng/ml in the tumour patients when 0.01 micrograms/ml hydrocortisone was added (P less than 0.05). At 10 micrograms/ml hydrocortisone suppressed IFN gamma production completely in both groups. Our results corroborate those investigations that showed no influence of the compound on T cell effector mechanisms. The attenuation of humoral immunophenomena, however, suggest a very specific point of action within the immune system by Ciamexone.


Bovine and human insulin activate CD8+-autoreactive CTL expressing both type 1 and type 2 cytokines in C57BL/6 mice.
CD8+ T cells down-regulate a variety of immune responses. For example, porcine and human insulin do not stimulate Abs in C57BL/6 mice because CD8+ T cells inhibit CD4+ helper T cells. By contrast, bovine insulin induces Ab in C57BL/6 mice, and removal of CD8+ T cells does not alter this response. This raises the question of whether porcine, but not bovine, insulin activates CD8+ T cells or whether both insulins activate CD8+ T cells but CD4+ helper T cells are differentially inhibited by them. In this study, we show that insulin-specific CD8+ CTL can be cultured from C57BL/6 mice primed with either bovine or human insulin in CFA. Thus, exogenous Ags, besides OVA, induce CD8+ CTL when administered in an adjuvant, suggesting this is a typical response. These CTL are H-2Kb restricted and produce IL-5, IL-10, IFN-gamma, and small amounts of IL-4, which is distinct from IFN-gamma and TNF-alpha that are typically secreted by virus-specific CTL. Moreover, the CTL primed with either bovine or human insulin recognize an A-chain peptide that is identical to the mouse insulin sequence. That foreign proteins, which are closely related to self-proteins, activated autoreactive, CD8+ T cells in vivo is a novel finding. It raises the possibility that self-reactive CTL may be activated by cross-reacting Ags and once activated they might participate in autoimmunity. These results also suggest that down-regulation of insulin-specific responses by autoreactive CD8+ T cells is most likely due to the differential sensitivity of bovine and human insulin-specific CD4+ T cells.


Metastatic Colon Cancer in an 18-Year-Old without Predisposing Factors.
While colorectal carcinoma is a common gastrointestinal cancer in adults, it is rare in pediatrics with an incidence of 1 : 1,000,000 and represents a fraction of neoplasms encountered in children. Malignant neoplasms represent a major cause of mortality in the pediatric age group. While presenting with weight loss, iron deficiency, rectal bleeding, abdominal pain, and change in bowel habits, or symptoms similar to acute appendicitis, the working diagnosis may be considered to be anorexia. This case illustrates the importance of considering colon cancer among other disease entities as a cause of unintentional weight loss in adolescents. While this is a rare occurrence in the pediatric population, significant unintentional weight loss with altered bowel habits should prompt a search for underlying malignancy-even in the absence of a positive family history or predisposing cancer syndromes.


Antifertility and profertility effects of the leaves and seeds of fluted pumpkin: Sperm quality, hormonal effects and histomorphological changes in the testes of experimental animal models.
Fluted pumpkin (FP; Telfairia occidentalis) is an edible vegetable, grown in West Africa, that is used in traditional medicine for its regulatory effects on the male gonads. Scientific articles concerning the effects of FP were identified by searching PubMed, PubChem, Scopus, Springer, ResearchGate, Google Scholar and Web of Science; this literature was to better understand the effects of FP seed (FPS) and leaf (FPL) extracts on the testes. Data showed that in experimental animals extracts of FPL and FPS at 1/100 of the lethal dose promoted testis regeneration and improved testosterone concentration and sperm quality, while at higher doses they had antifertility effects. Several extracts of FPS and FPL, including ethanol, aqueous, methanol and hydroethanolic, had protective effects on the testes of study animals at lower doses (≥ 50 mg/kg body weight), but at higher doses (≥ 200 mg/kg body weight) they inhibited hormone synthesis, sperm quality and histomorphological structure, under both normal and disease conditions. The posttreatment effects of FPS on the gonads were reversible in young mature rats and FPS had slight systemic toxic effects. Although, there are inconsistencies in some of the published results, the current evidence suggests that FPS and FPL have both profertility and reversible antifertility effects in experimental animals.


A Systematic Review and Meta-analysis on the Prevalence of HBsAg in Health Care Workers from Eastern Mediterranean and Middle Eastern Countries.
The present study aimed to determine the prevalence of HBsAg in Health Care Workers (HCWs) in Eastern Mediterranean Region Office (EMRO) and Middle Eastern countries from 2000 to 2016. In a meta-analysis study, the databases of PubMed, ISI, Ovid, Scopus, Google Scholar, and Persian databases were searched for relevant articles on the prevalence of HBsAg in HCWs in EMRO and Middle Eastern countries. Homogeneity was assessed based on Cochran's Q-test results. A total of 43 articles (110,179 people) were included. The pooled prevalence of HBsAg in HCWs of EMRO and Middle East countries was found 2.77% (95%CI: 2.64-2.83). The specific prevalence of HBsAg was 2.84% (95% CI: 2.6-3.11) in EMRO and 2.22% (95%CI: 2.13-2.31) in Middle Eastern countries. The highest and lowest prevalence rates of HBsAg among HCWs for countries with more than one study were 6.85% (95% CI: 5.74%-8.16%) in Sudan and 1.00% (95% CI: 0.94%-1.07%) in Turkey, respectively. The trends of HBsAg prevalence among HCWs decreased from 2000 to 2016. Based on the World Health Organization classification of HBV prevalence, intermediate HBsAg prevalence rates were detected in HCWs of EMRO and Middle East countries during 2000-2016.


Soil Landscape Pattern Changes in Response to Rural Anthropogenic Activity across Tiaoxi Watershed, China.
Soil sealing (loss of soil resources due to extensive land covering for the purpose of house building, road construction etc.) and subsequent soil landscape pattern changes constitute typical environmental problems in many places worldwide. Previous studies concentrated on soil sealing in urbanized regions, whereas rural areas have not been given sufficient attention. Accordingly, this paper studies soil landscape pattern dynamics (i.e., landscape pattern changes in response to rural anthropogenic activities) in the Tiaoxi watershed (Zhejiang province, eastern China), in which surface sealing is by far the predominant component of human forcing with respect to environmental change. A novel approach of quantifying the impacts of rural anthropogenic activities on soil resources is presented. Specifically, quantitative relationships were derived between five soil landscape pattern metrics (patch density, edge density, shape index, Shannon's diversity index and aggregation index) and three rural anthropogenic activity indicators (anthropogenic activity intensity, distance to towns, and distance to roads) at two landscape block scales (3 and 5 km) between 1985 and 2010. The results showed that the Tiaoxi watershed experienced extensive rural settlement expansion and high rates of soil sealing. Soil landscapes became more fragmented, more irregular, more isolated, and less diverse. Relationships between soil landscape pattern changes and rural anthropogenic activities differed with the scale (spatial and temporal) and variable considered. In particular, the anthropogenic activity intensity was found to be the most important indicator explaining social development intensity, whereas the other two proximity indicators had a significant impact at certain temporal interval. In combination with scale effects, spatial dependency (correlation) was shown to play a key role that should be carefully taken into consideration in any relevant environmental study. Overall, the findings of this work suggest that soil sealing can be a critical human forcing issue with considerable consequences deserving serious attention by the experts, the public and the government alike.


Olanzapine Attenuates Mechanical Allodynia in a Rat Model of Partial Sciatic Nerve Ligation.
Neuropathic pain is a global clinical problem; nevertheless, nerve injury treatment methods remain limited. Olanzapine has antinociceptive and anti-nueropathic properties; however, its preventive effects have not been assessed in nerve injury models. We prepared a partial sciatic nerve ligation (Seltzer model) or sham-operated model in male Sprague-Dawley rats under isoflurane anesthesia. In a pre-treatment study, we administered olanzapine (10 mg/kg) intraperitoneally 1 h before nerve ligation. In post-treatment and dose-dependent studies, we injected 3 different doses of olanzapine intraperitoneally 1 h after nerve ligation. Mechanical allodynia was measured before and 7 days after surgery. Immunohistochemical analysis using anti-Iba-1 antibody was used to assess the effect of olanzapine at the spinal level. In the pre-treatment study, median withdrawal thresholds of the normal saline groups were significantly lower than those of the sham-operated groups; however, those of the olanzapine (10 mg/kg) and sham-operated groups were not different. In the post-treatment and dose-dependent studies, the median withdrawal thresholds of the olanzapine (2.5 mg/kg) and normal saline groups were not different; however, those of the olanzapine (10 and 50 mg/kg) groups were significantly higher than those of the normal saline groups. Olanzapine did not have a significant effect on the density of Iba-1 staining. Olanzapine attenuated mechanical allodynia dose-dependently in the Seltzer model. This anti-allodynic effect of olanzapine was observed even when injected 1 h after nerve ligation. This effect of olanzapine appeared to be unrelated to microglia activation in the ipsilateral dorsal horn of the lumbar spinal cord.


Efficient initiation of DNA replication in eukaryotes requires Dpb11/TopBP1-GINS interaction.
Dpb11/Cut5/TopBP1 is evolutionarily conserved and is essential for the initiation of DNA replication in eukaryotes. The Dpb11 of the budding yeast Saccharomyces cerevisiae has four BRCT domains (BRCT1 to -4). The N-terminal pair (BRCT1 and -2) and the C-terminal pair (BRCT3 and -4) bind to cyclin-dependent kinase (CDK)-phosphorylated Sld3 and Sld2, respectively. These phosphorylation-dependent interactions trigger the initiation of DNA replication. BRCT1 and -2 and BRCT3 and -4 of Dpb11 are separated by a short stretch of ~100 amino acids. It is unknown whether this inter-BRCT region functions in DNA replication. Here, we showed that the inter-BRCT region is a GINS interaction domain that is essential for cell growth and that mutations in this domain cause replication defects in budding yeast. We found the corresponding region in the vertebrate ortholog, TopBP1, and showed that the corresponding region also interacts with GINS and is required for efficient DNA replication. We propose that the inter-BRCT region of Dpb11 is a functionally conserved GINS interaction domain that is important for the initiation of DNA replication in eukaryotes.


[Aggregation and swelling of brain synaptic vesicles in rats].
Mg-ATPase (1 mM) induces a decrease in the intensity of light scattering (I1) at 620 nm of rat brain synaptic vesicles (SV) suspended in sucrose, with this decrease being indicative of the swelling of the vesicles. The Mg-ATPase-induced swelling appears to be associated with the function of H+-ATPase of SV membranes, since it is completely abolished by the proton pump blocker dicyclohexylcarbodiimide and the protonophore carbonylcyanide m-chlorophenylhydrazone. The Mg-ATPase-induced swelling was enhanced in the presence of the permeable anion Cl- (in the range of 25-50 mM KCl). Ca2+ (and Mg2+) at high concentrations (0.1-1.0 mM) cause aggregation of the SV as measured by changes in the I1. Colchicine and cytochalasin do not affect SV swelling and aggregation whereas Mg-ATP (1 mM) lowers aggregation caused by Ca2+ (1 mM).


Genome-wide association study of bronchopulmonary dysplasia: a potential role for variants near the CRP gene.
Bronchopulmonary dysplasia (BPD), the main consequence of prematurity, has a significant heritability, but little is known about predisposing genes. The aim of this study was to identify gene loci predisposing infants to BPD. The initial genome-wide association study (GWAS) included 174 Finnish preterm infants of gestational age 24-30 weeks. Thereafter, the most promising single-nucleotide polymorphisms (SNPs) associated with BPD were genotyped in both Finnish (n = 555) and non-Finnish (n = 388) replication cohorts. Finally, plasma CRP levels from the first week of life and the risk of BPD were assessed. SNP rs11265269, flanking the CRP gene, showed the strongest signal in GWAS (odds ratio [OR] 3.2, p = 3.4 × 10-6). This association was nominally replicated in Finnish and French African populations. A number of other SNPs in the CRP region, including rs3093059, had nominal associations with BPD. During the first week of life the elevated plasma levels of CRP predicted the risk of BPD (OR 3.4, p = 2.9 × 10-4) and the SNP rs3093059 associated nominally with plasma CRP levels. Finally, SNP rs11265269 was identified as a risk factor of BPD (OR 1.8, p = 5.3 × 10-5), independently of the robust antenatal risk factors. As such, in BPD, a potential role for variants near CRP gene is proposed.


Small-vessel radiography in situ with monochromatic synchrotron radiation.
To evaluate the usefulness of a radiographic system with monochromatic synchrotron radiation to depict small vessels and peripheral secretory ducts. Radiography of various organs was tested in 14 anesthetized dogs and pancreatography was performed in an excised human pancreas by using the following system: monochromatic synchrotron radiation with an energy level just above the k absorption edge of iodine as an x-ray source and a high-definition TV system with a high-light-sensitivity image pick-up tube camera coupled with a fluorescent screen as a detector. This system allowed depiction of small vessels (diameter < 50-100 microns) of the heart (penetrating transmural artery), brain (perforating arteries that arise directly in the circle of Willis), and intestinal organs (vasa recta and their submucosal communications) and of small branches (down to the fifth order) of the pancreatic duct. The synchrotron radiation system may be useful for evaluating microcirculatory disorders and early-stage malignant tumors in various human organs.


Eveningness is associated with higher risk-taking in dangerous driving situations.
Inclination toward eveningness is often associated with risky behavior. But the existing studies are scarce, inconsistent and usually limited to self-reported measures. We sought to investigate in young adults whether morningness-eveningness is associated with risky behavior in dangerous driving situations, with self-reported drunk driving and with alcohol consumption. Results show that, indeed, inclination toward eveningness is associated with these risky behaviors. We also demonstrate a link between morningness-eveningness and sensation seeking. Therefore, young adults with a tendency toward eveningness might be more at risk to face negative consequences of alcohol abuse or to be involved in a road accident.


Direct atomic force microscope imaging of EcoRI endonuclease site specifically bound to plasmid DNA molecules.
Direct imaging with the atomic force microscope has been used to identify specific nucleotide sequences in plasmid DNA molecules. This was accomplished using EcoRI (Gln-111), a mutant of the restriction enzyme that has a 1000-fold greater binding affinity than the wild-type enzyme but with cleavage rate constants reduced by a factor of 10(4). ScaI-linearized plasmids with single (pBS+) and double (pGEM-luc and pSV-beta-galactosidase) EcoRI recognition sites were imaged, and the bound enzyme was localized to a 50- to 100-nt resolution. The high affinity for the EcoRI binding site exhibited by this mutant endonuclease, coupled with an observed low level of nonspecific binding, should prove valuable for physically mapping large DNA clones by direct atomic force microscope imaging.


Validity and Reliability of Willingness-to-Pay Estimates: Evidence from Two Overlapping Discrete-Choice Experiments.
Discrete-choice experiments (DCEs), while becoming increasingly popular, have rarely been tested for validity and reliability. To address the issues of validity and reliability of willingness-to-accept (WTA) values obtained from DCEs. In particular, to examine whether differences in the attribute set describing a hypothetical product have an influence on preferences and willingness-to-pay (WTP) values of respondents. Two DCEs were designed, featuring hypothetical insurance contracts for Swiss healthcare. The contract attributes were pre-selected in expert sessions with representatives of the Swiss healthcare system, and their relevance was checked in a pre-test. Experiment A contained rather radical health system reform options, while experiment B concentrated on more familiar elements such as co-payment and the benefit catalogue. Three attributes were present in both experiments: delayed access to innovation ('innovation'), restricted drug benefit ('generics'), and the change in the monthly premium ('premium'). The issue to be addressed was whether WTA values for the overlapping attributes were similar, even though they were embedded in widely differing choice sets.Two representative telephone surveys with 1000 people aged >25 years were conducted independently in the German and French parts of Switzerland during September 2003. Socioeconomic variables collected included age, sex, education, total household income, place of residence, occupation, and household size. Three models were estimated (a simple linear model, a model allowing interaction of the price attribute with socioeconomic characteristics, and a model with a full set of interaction terms). The socioeconomic characteristics of the two samples were very similar. Theoretical validity tends to receive empirical support in both experiments in all cases where economic theory makes predictions concerning differences between socioeconomic groups. However, a systematic inappropriate influence on measured WTA seems to be present in at least one experiment. This is likely to be experiment A, in which respondents were far less familiar with proposed alternatives than in experiment B. Measuring preferences for major, little-known innovations in a reliable way seems to present particular challenges for experimental research.


Treatment of experimental cryptococcal meningitis with fluconazole: impact of dose and addition of flucytosine on mycologic and pathophysiologic outcome.
Fluconazole is effective in the therapy of cryptococcal meningitis in patients with AIDS. The optimal dosage of fluconazole and the impact of combination with flucytosine are not known. In this study, rabbits with experimental cryptococcal meningitis were given fluconazole at low, intermediate, or high dose or in combination with a low or intermediate dose of flucytosine. Serial cerebrospinal fluid (CSF) examinations showed that all three doses of fluconazole and low-dose fluconazole in combination with intermediate-dose flucytosine were effective in reducing CSF cryptococcal titer, lactate, white blood cell count, and cryptococcal antigen (CRAG) titers. The intermediate and high doses of fluconazole reduced CSF fungal (P < .05) and CRAG (P < .001) titers earlier than low-dose fluconazole alone or in combination with flucytosine. Only the highest dose of fluconazole reduced brain edema after 7 days. In this model of cryptococcal meningitis, there was evidence of a dose response with fluconazole but no in vivo synergism with flucytosine.


Stimulation of intrachromosomal homologous recombination in human cells by electroporation with site-specific endonucleases.
In somatic mammalian cells, homologous recombination is a rare event. To study the effects of chromosomal breaks on frequency of homologous recombination, site-specific endonucleases were introduced into human cells by electroporation. Cell lines with a partial duplication within the HPRT (hypoxanthine phosphoribosyltransferase) gene were created through gene targeting. Homologous intrachromosomal recombination between the repeated regions of the gene can reconstruct a functioning, wild-type gene. Treatment of these cells with the restriction endonuclease Xba I, which has a recognition site within the repeated region of HPRT homology, increased the frequency or homologous recombination bv more than 10-fold. Recombination frequency was similarly increased by treatment with the rare-cutting yeast endonuclease PI-Sce I when a cleavage site was placed within the repeated region of HPRT. In contrast, four restriction enzymes that cut at positions either outside of the repeated regions or between them produced no change in recombination frequency. The results suggest that homologous recombination between intrachromosomal repeats can be specifically initiated by a double-strand break occurring within regions of homology, consistent with the predictions of a model.


[Molecular diagnosis and gene therapy for Gaucher disease].
Gaucher disease is the most prevalent lysosomal storage disease. It is caused by deficient activity of a lysosomal enzyme known as glucocerebrosidase, also called glucosylceramidase, resulting from mutations in the gene encoding the enzyme. The numerous mutations in glucocerebrosidase gene from patients were reported and the correlation of phenotype and genotype were studied. However, a given genotype cannot be uniquely correlated with a specific phenotype. In the therapeutic point of view, two successful treatment were developed based on the correction of the enzyme deficiency in macrophages. These are bone marrow transplantation and enzyme infusion therapy. Although patients have responded to these two therapies, inherent problems limit their application. Thus, these problems makes Gaucher disease an excellent candidate for therapy based on gene transfer to hematopoietic cells. We succeeded in efficient transduction and sustained high expression of glucocerebrosidase gene in mouse hematopoietic stem cells and macrophages from long term reconstituted mice. The results of our study strengthen the rationale for gene therapy as a treatment for Gaucher disease.


The concept of hope in nursing 4: hope and gerontological nursing.
This article is the fourth in a series of six that explores the nature of hope, reviews the existing theoretical and empirical work in several discrete areas of nursing and provides case studies to illustrate the role that hope plays in clinical situations. This article focuses on hope within the specialty of gerontological nursing. To date, most of the hope research, using qualitative and quantitative methodologies, has focused on well and chronically ill young or middle-aged adults; very few studies have examined hope in the older adult and specifically how older adults maintain hope in spite of multiple losses and/or changes. Hope has been studied in several elderly populations, including grieving widow(er)s, community-based older persons, persons living in long term care facilities and elders recruited from senior citizen centres. Research findings provide initial direction for the assessment of hope, and a reference for evaluating the effectiveness of these strategies. There remains, however, a need for continuing research in the area of hope in the older adult that is multifaceted and applicable to both practice and education.