Estrogenic chemicals often leach from BPA-free plastic products that are replacements for BPA-containing polycarbonate products.
Xenobiotic chemicals with estrogenic activity (EA), such as bisphenol A (BPA), have been reported to have potential adverse health effects in mammals, including humans, especially in fetal and infant stages. Concerns about safety have caused many manufacturers to use alternatives to polycarbonate (PC) resins to make hard and clear, reusable, plastic products that do not leach BPA. However, no study has focused on whether such BPA-free PC-replacement products, chosen for their perceived higher safety, especially for babies, also release other chemicals that have EA. We used two, well-established, mammalian cell-based, assays (MCF-7 and BG1Luc) to assess the EA of chemicals that leached into over 1000 saline or ethanol extracts of 50 unstressed or stressed (autoclaving, microwaving, and UV radiation) BPA-free PC-replacement products. An EA antagonist, ICI 182,780, was used to confirm that agonist activity in leachates was due to chemicals that activated the mammalian estrogen receptor. Many unstressed and stressed, PC-replacement-products made from acrylic, polystyrene, polyethersulfone, and Tritan™ resins leached chemicals with EA, including products made for use by babies. Exposure to various forms of UV radiation often increased the leaching of chemicals with EA. In contrast, some BPA-free PC-replacement products made from glycol-modified polyethylene terephthalate or cyclic olefin polymer or co-polymer resins did not release chemicals with detectable EA under any conditions tested. This hazard assessment survey showed that many BPA-free PC- replacement products still leached chemicals having significant levels of EA, as did BPA-containing PC counterparts they were meant to replace. That is, BPA-free did not mean EA-free. However, this study also showed that some PC-replacement products did not leach chemicals having significant levels of EA. That is, EA-free PC-replacement products could be made in commercial quantities at prices that compete with PC-replacement products that were not BPA-free. Since plastic products often have advantages (price, weight, shatter-resistance, etc.) compared to other materials such as steel or glass, it is not necessary to forgo those advantages to avoid release into foodstuffs or the environment of chemicals having EA that may have potential adverse effects on our health or the health of future generations.


Despite high levels of expression in thymic epithelial cells, miR-181a1 and miR-181b1 are not required for thymic development.
MicroRNAs (miRNAs) have been shown to be key modulators of post-transcriptional gene silencing in many cellular processes. In previous studies designed to understand the role of miRNAs in thymic development, we globally deleted miRNA exclusively in thymic epithelial cells (TECs), which are critical in thymic selection. This resulted in the loss of stromal cells that instruct T cell lineage commitment and affect thymocyte positive selection, required for mature T cell development. Since murine miR-181 is expressed in the thymus and miR-181 deficiency disrupts thymocyte development, we first quantified and thereby demonstrated that miR181a1 and miR181b1 are expressed in purified TECs. By generating mice with TEC targeted loss of miR-181a1 and miR-181b1 expression, we observed that neither TEC cellularity nor thymocyte number nor differentiation was adversely affected. Thus, disrupted thymopoiesis in miR-181 deficient mice was not due to miR-181 loss of expression in TECs. Importantly, in mice with restricted TEC deficiency of miR-181a1 and miR-181b1, there were similar numbers of mature T cells in the periphery in regards to frequencies, differentiation, and function as compared to controls. Moreover miR-181a1 and miR-181b1 were not required for maintenance of thymus integrity over time, as thymic involution was not accelerated in gene-targeted mice. Taken together our data indicate that miR-181a1 and miR-181b1 are dispensable for TEC differentiation, their control of thymocyte development and mature T cell export to and homeostasis within the periphery.


Chondrogenic differentiation of mesenchymal stem cells in a hydrogel system based on an enzymatically crosslinked tyramine derivative of hyaluronan.
Hyaluronan-based tissue substitutes are promising materials in cartilage reconstruction surgery. Herein, the chondrogenesis of human mesenchymal stem cells (MSC) in a hydrogel based on a tyramine derivative of hyaluronan crosslinked by hydrogen peroxidase (HA-TA) was evaluated. Human MSC seeded in the scaffold were incubated in standard chondrogenic medium and medium enriched with bone morphogenetic protein-6 (BMP6). Cell viability, the gene expression of selected markers (collagen type II, aggrecan, SOX9, collagen type X, and osteopontin), and the histological characteristics were examined during three weeks of in vitro cultivation. The tissue reaction of both unseeded and MSC seeded HA-TA scaffolds were tested in vivo after subcutaneous application in rats for 12 weeks. The data showed that cells resisted the process of crosslinking and remained viable for the whole time while exhibiting changes in cell organization. Human MSC cultivated in HA-TA hydrogel expressed genes of both chondrogenic and osteogenic differentiation and the addition of BMP6 revealed a tendency to potentiate both processes. Histological analysis of HA-TA in vivo implants did not reveal a chronic inflammatory reaction. In both cases, in vivo HA-TA implants were continuously degraded and MSC-seeded hydrogels tended to form clusters similar to in vitro samples. In conclusion, MSC chondrogenic differentiation may proceed in a HA-TA scaffold that is biocompatible. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3523-3530, 2014.


Combination therapy in hypertension.
The goal of antihypertensive therapy is to abolish the risks associated with blood pressure (BP) elevation without adversely affecting quality of life. Drug selection is based on efficacy in lowering BP and in reducing cardiovascular (CV) end points including stroke, myocardial infarction, and heart failure. Although the choice of initial drug therapy exerts some effect on long-term outcomes, it is evident that BP reduction per se is the primary determinant of CV risk reduction. Available data suggest that at least 75% of patients will require combination therapy to achieve contemporary BP targets, and increasing emphasis is being placed on the practical tasks involved in consistently achieving and maintaining goal BP in clinical practice. It is within this context that the American Society of Hypertension presents this Position Paper on Combination Therapy for Hypertension. It will address the scientific basis of combination therapy, present the pharmacologic rationale for choosing specific drug combinations, and review patient selection criteria for initial and secondary use. The advantages and disadvantages of single pill (fixed) drug combinations, and the implications of recent clinical trials involving specific combination strategies will also be discussed.


Clinical Significance of Expression of Nephroblastoma Overexpressed (NOV) in Patients with Colorectal Cancer.
The nephroblastoma overexpressed (NOV) gene, which belongs to the cysteine-rich, angiogenic inducer 61/connective tissue growth factor/nephroblastoma overexpressed (CCN) family, is located in the 8q24 region and promotes migration and invasiveness in several types of malignancies. We explored the clinical significance of NOV expression in colorectal cancer (CRC). NOV expression in CRC specimens and CRC cell lines were evaluated. The association between the clinicopathlogical factors and NOV mRNA expression of tumor tissues was assessed in 126 patients with CRC. We assessed the relationships between NOV expression and overall survival in public databases. We performed overexpression experiments in vitro. CRC specimens and CRC cell lines showed high NOV expression. High NOV mRNA expression was correlated with poorer overall survival and higher Union for International Cancer Control (UICC) T factor. In public databases, high NOV expression was associated with poorer prognoses. Overexpression of NOV promoted invasiveness of CRC cells. NOV may be an indicator of poor prognosis and a therapeutic target in CRC.


A cardiac mitochondrial cAMP signaling pathway regulates calcium accumulation, permeability transition and cell death.
Although cardiac cytosolic cyclic 3',5'-adenosine monophosphate (cAMP) regulates multiple processes, such as beating, contractility, metabolism and apoptosis, little is known yet on the role of this second messenger within cardiac mitochondria. Using cellular and subcellular approaches, we demonstrate here the local expression of several actors of cAMP signaling within cardiac mitochondria, namely a truncated form of soluble AC (sACt) and the exchange protein directly activated by cAMP 1 (Epac1), and show a protective role for sACt against cell death, apoptosis as well as necrosis in primary cardiomyocytes. Upon stimulation with bicarbonate (HCO3(-)) and Ca(2+), sACt produces cAMP, which in turn stimulates oxygen consumption, increases the mitochondrial membrane potential (ΔΨm) and ATP production. cAMP is rate limiting for matrix Ca(2+) entry via Epac1 and the mitochondrial calcium uniporter and, as a consequence, prevents mitochondrial permeability transition (MPT). The mitochondrial cAMP effects involve neither protein kinase A, Epac2 nor the mitochondrial Na(+)/Ca(2+) exchanger. In addition, in mitochondria isolated from failing rat hearts, stimulation of the mitochondrial cAMP pathway by HCO3(-) rescued the sensitization of mitochondria to Ca(2+)-induced MPT. Thus, our study identifies a link between mitochondrial cAMP, mitochondrial metabolism and cell death in the heart, which is independent of cytosolic cAMP signaling. Our results might have implications for therapeutic prevention of cell death in cardiac pathologies.


Explanatory models of diabetes: patient practitioner variation.
Most cases of diabetes, a complex disorder that requires many lifestyle changes, can be controlled if persons adhere to their prescribed regimen. However, compliance is difficult to attain. Differences in explanatory models between client and practitioner have been suggested as one reason for non-compliance in several disorders. In this ethnographic investigation, individual explanatory models were elicited from persons with diabetes and from health professionals working with these patients. Professionals described models of diabetes in general and their model of a particular patient's diabetes. A composite professional model was constructed and compared with each of the patients' models. The models were most congruent regarding treatment. Etiology, pathophysiology, and severity had less congruence, and time and mode of symptom onset were least congruent. The Spearman correlation coefficient showed a positive but non-significant association of explanatory model congruence between professionals and patients with normal glycosylated hemoglobin levels. Patients and professionals seem to emphasize different domains; patients emphasized difficulties in the social domain and the impact of diabetes on their lives while staff saw diabetes primarily as a pathophysiological problem with impact on patients' physical bodies. This study's importance rests on its clear articulation of significant differences between patients' and staffs' models even when they are similar in demographic characteristics.


CT virtual endoscopy of the stomach: comparison study with gastric fiberscopy.
We evaluated the value of computed tomographic (CT) gastric virtual endoscopy (VE) by comparing it with real gastric fiberscopy (GF). Sixty-six VE findings in 63 patients were compared with lesions found with GF. After a GF examination, the examinees were given bubble-making granules for the stomach and placed in a supine position on the CT table. Abdominal CT slices of 1 mm thickness were obtained by multidetector row CT (Siemens Sensation 16) and reconstructed to VE images with volume and surface rendering on the same day. VE images were successful in showing subtle alterations in the gastric mucosal folds. Gastric cancer, polyps, ulcers, erosions, and gastritis were clearly visualized. Comparison studies between VE and GF (as the gold standard) showed a sensitivity of 92.7% and a specificity of 90.9%. Original CT images were also useful in detecting lesions in other abdominal organs such as the liver, adrenal gland, and kidney. Because VE is a good noninvasive screening method, except for minimal radiation exposure, it is expected to become a new technique for stomach examinations.


Use of an artificial lymphatic system during carboplatin infusion to improve canine osteosarcoma blood flow and clinical response.
The artificial lymphatic system (ALS), a mechanical system designed to reduce increased interstitial fluid pressure in solid tumors and enhance the delivery of chemotherapy, was evaluated within a randomized clinical trial treating spontaneously occurring canine appendicular osteosarcoma (OS), a tumor similar to its human OS counterpart. An ALS was investigated for its ability to increase OS blood flow and increase uptake of intravenously administered carboplatin. Blood flow increased by 314% in tumors with active ALS drains versus 126% in control tumors (P < .03). Tumor carboplatin uptake increased by 51% after drain activation (P = .07). Microvascular density (MVD) was measured in tumors after surgical amputation and in corresponding bone regions in a cohort of normal dogs. The OS tumors had equivalent MVD as normal bone, and MVD was higher in the humerus than the femur (P < .03) in both tumor and normal bone. Median survival between the ALS-treated and control cohorts was not different despite increased drug uptake or ALS manipulation. Compared with historic controls, ALS drain insertion into tumors to reduce interstitial fluid pressure did not worsen the prognosis. The findings in canine spontaneously occurring OS indicate that an ALS may be of value as a chemotherapy adjunct for enhancing the delivery of chemotherapy to tumor interstitium.


DNA topoisomerase IIalpha is required for RNA polymerase II transcription on chromatin templates.
In the nucleus of the cell, core RNA polymerase II (pol II) is associated with a large complex called the pol II holoenzyme (holo-pol). Transcription by core pol II in vitro on nucleosomal templates is repressed compared with that on templates of histone-free naked DNA. We found that the transcriptional activity of holo-pol, in contrast to that of core pol II, is not markedly repressed on chromatin templates. We refer to this property of holo-pol as chromatin-dependent coactivation (CDC). Here we show that DNA topoisomerase IIalpha is associated with the holo-pol and is a required component of CDC. Etoposide and ICRF-193, specific inhibitors of topoisomerase II, blocked transcription on chromatin templates, but did not affect transcription on naked templates. Addition of purified topoisomerase IIalpha reconstituted CDC activity in reactions with core pol II. These findings suggest that transcription on chromatin templates results in the accumulation of superhelical tension, making the relaxation activity of topoisomerase II essential for productive RNA synthesis on nucleosomal DNA.


Ethnicity, consanguinity, and genetic architecture of hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is characterized by phenotypic heterogeneity that is partly explained by the diversity of genetic variants contributing to disease. Accurate interpretation of these variants constitutes a major challenge for diagnosis and implementing precision medicine, especially in understudied populations. The aim is to define the genetic architecture of HCM in North African cohorts with high consanguinity using ancestry-matched cases and controls. Prospective Egyptian patients (n = 514) and controls (n = 400) underwent clinical phenotyping and genetic testing. Rare variants in 13 validated HCM genes were classified according to standard clinical guidelines and compared with a prospective HCM cohort of majority European ancestry (n = 684). A higher prevalence of homozygous variants was observed in Egyptian patients (4.1% vs. 0.1%, P = 2 × 10-7), with variants in the minor HCM genes MYL2, MYL3, and CSRP3 more likely to present in homozygosity than the major genes, suggesting these variants are less penetrant in heterozygosity. Biallelic variants in the recessive HCM gene TRIM63 were detected in 2.1% of patients (five-fold greater than European patients), highlighting the importance of recessive inheritance in consanguineous populations. Finally, rare variants in Egyptian HCM patients were less likely to be classified as (likely) pathogenic compared with Europeans (40.8% vs. 61.6%, P = 1.6 × 10-5) due to the underrepresentation of Middle Eastern populations in current reference resources. This proportion increased to 53.3% after incorporating methods that leverage new ancestry-matched controls presented here. Studying consanguineous populations reveals novel insights with relevance to genetic testing and our understanding of the genetic architecture of HCM.


Low-grade inflammation is negatively associated with physical Health-Related Quality of Life in healthy individuals: Results from The Danish Blood Donor Study (DBDS).
Health-Related Quality of Life (HRQL) represent individuals' subjective assessment of their mental and physical well-being, and is highly predictive of future health. C-reactive protein (CRP) is a well-established marker of inflammation. Low-grade inflammation (LGI), defined as slightly increased CRP levels, is associated with increased risk of several diseases. LGI may reflect subclinical pathology, which could affect individual's subjective health assessment. This study aimed to examine whether LGI has an independent impact on self-reported health or rather is a mediator of a confounder in a large population of healthy individuals. Plasma CRP levels were measured in 17,024 participants from the Danish Blood Donor Study (DBDS). All participants completed a standard questionnaire including smoking status, and the 12-item short-form health survey (SF-12), which is a widely used scale for HRQL. SF-12 is reported as a mental (MCS) and physical (PCS) score. The relationship between LGI (defined as a plasma CRP level between 3 mg/L and 10 mg/L) and MCS or PCS was explored by mediation analysis and adjusted multivariable linear regression analysis. Multiple imputation modelling was used to remedy missing values. The analyses were stratified according to sex and use of combined oral contraception (OC). In the study, 1,542 (10.3%) participants had LGI. PCS was associated with LGI in all strata, i.e. women using OC: RC = -0.36 points lower PCS in participants with LGI vs no LGI, CI: -0.94 to -0.19, women not using OC: RC = -0.63, CI: -1.05 to -0.21 and men: RC = -0.76, CI: -1.10 to -0.42. But LGI had no impact on MCS. Predictors of lower PCS included obesity, current smoking, and waist circumference in all strata. Physical activity in leisure time was the only factor positively associated with PCS. Age and physical activity in leisure time was associated with increased MCS in all strata whereas current smoking was the only strong predictor of a reduction in MCS. Only a small effect of smoking on PCS was mediated through LGI. In this population of healthy individuals, LGI had independent impact on lower self-rated physical health score in HRQL in both sexes, but was not associated with self-rated mental health score. A small and significant effect of smoking on physical health score was mediated through LGI.


The dynamics of response as measured by multiple composite outcome tools in the TIght COntrol of inflammation in early Psoriatic Arthritis (TICOPA) trial.
We aimed to evaluate the dynamics of treatment response with different composite measures in the TIght COntrol of inflammation in early Psoriatic Arthritis (TICOPA) trial. Participants with early disease-modifying antirheumatic drug-naïve psoriatic arthritis (PsA) were randomised 1:1 to either tight control (TC; 4 weekly review with therapy escalation if criteria not met) or standard care (SC; 12 weekly review). We calculated modified versions of the Psoriatic ArthritiS Disease Activity Score (PASDAS), Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA) Composite scorE (GRACE) and Composite Psoriatic Disease Activity Index (CPDAI) at baseline and 12 weekly to 48 weeks by blinded assessor. For missing data, we used the last observation carried forward. Comparison between groups was made by analysis of covariance and comparison of area under the curve (AUC). 206 people were randomised to TC (n=101) or SC (n=105). Significant differences between treatment groups were seen (p<0.0001 for all composite measures). AUC analysis demonstrated a significant difference between groups for the PASDAS but not GRACE and CPDAI. For participants with oligoarthritis, a significant difference between groups was seen for each measure, although the significance levels were greatly diminished (PASDAS, p=0.04; GRACE p=0.01; CPDAI p=0.04). For oligoarthritis using AUC analysis, none of the measures could distinguish between groups. Composite measures of disease activity were able to distinguish between TICOPA treatment arms, although differences were diminished for those with oligoarthritis. Further data are needed to inform the preferred composite measure for use as the primary outcome in PsA trials. ClinicalTrials.gov (NCT01106079) and ISCRCTN registry (ISCRCTN30147736).


Doxycycline for outpatient-treated acute exacerbations of COPD: a randomised double-blind placebo-controlled trial.
Antibiotics do not reduce mortality or short-term treatment non-response in patients receiving treatment for acute exacerbations of COPD in an outpatient setting. However, the long-term effects of antibiotics are unknown. The aim of this study was to investigate if the antibiotic doxycycline added to the oral corticosteroid prednisolone prolongs time to next exacerbation in patients with COPD receiving treatment for an exacerbation in the outpatient setting. In this randomised double-blind placebo-controlled trial, we recruited a cohort of patients with COPD from outpatient clinics of nine teaching hospitals and three primary care centres in the Netherlands. Inclusion criteria were an age of at least 45 years, a smoking history of at least 10 pack-years, mild-to-severe COPD (Global Initiative of Chronic Obstructive Lung Disease [GOLD] stage 1-3), and at least one exacerbation during the past 3 years. Exclusion criteria were poor mastery of the Dutch language, poor cognitive functioning, known allergy to doxycycline, pregnancy, and a life expectancy of shorter than 1 month. If a participant had an exacerbation, we randomly assigned them (1:1; with permuted blocks of variable sizes [ranging from two to ten]; stratified by GOLD stage 1-2 vs 3) to a 7 day course of oral doxycycline 100 mg daily (200 mg on the first day) or placebo. Exclusion criteria for randomisation were fever, admission to hospital, and current use of antibiotics or use within the previous 3 weeks. Patients in both groups received a 10 day course of 30 mg oral prednisolone daily. Patients, investigators, and those assessing outcomes were masked to treatment assignment. The primary outcome was time to next exacerbation in all randomly allocated patients except for those incorrectly randomly allocated who did not meet the inclusion criteria or met the exclusion criteria. This trial is registered with the Netherlands Trial Register, number NTR2499. Between Dec 22, 2010, and Aug 6, 2013, we randomly allocated 305 (34%) patients from the cohort of 887 patients to doxycycline (152 [50%]) or placebo (153 [50%]), excluding four (1%) patients (two [1%] from each group) who were incorrectly randomly allocated from the analysis. 257 (85%) of 301 patients had a next exacerbation (131 [87%] of 150 in the doxycycline group vs 126 [83%] of 151 in the placebo group). Median time to next exacerbation was 148 days (95% CI 95-200) in the doxycycline group compared with 161 days (118-211) in the placebo group (hazard ratio 1·01 [95% CI 0·79-1·31]; p=0·91). We did not note any significant differences between groups in the frequency of adverse events during the first 2 weeks after randomisation (47 [31%] of 150 in the doxycycline group vs 53 [35%] of 151 in the placebo group; p=0·54) or in serious adverse events during the 2 years of follow-up (42 [28%] vs 43 [29%]; p=1). In patients with mild-to-severe COPD receiving treatment for an exacerbation in an outpatient setting, the antibiotic doxycycline added to the oral corticosteroid prednisolone did not prolong time to next exacerbation compared with prednisolone alone. These findings do not support prescription of antibiotics for COPD exacerbations in an outpatient setting. Netherlands Organization for Health Research and Development.


Isolation and characterization of Rhodovulum strictum sp. nov. and some other purple nonsulfur bacteria from colored blooms in tidal and seawater pools.
Several strains of phototrophic purple nonsulfur bacteria were isolated from colored blooms occurring in tidal and seawater pools in Japan. All of these isolates had ovoid to rod-shaped cells that were motile by means of single polar flagella and contained vesicular intracytoplasmic membranes together with bacteriochlorophyll a and carotenoids of the spheroidene series. They produced ubiquinone 10 as the major quinone and contained straight-chain fatty acids, with C18:1 predominating. They were mesophilic, halophilic, and photoheterotrophic, utilized sulfide and thiosulfate as electron donors for phototrophic growth, and photoassimilated a wide variety of organic compounds as carbon sources. Our results suggested that all of these isolates are members of the recently described genus Rhodovulum. The isolates were classified into four groups (designated groups I through IV) on the basis of phenotypic and genotypic data. The group I isolates, which were the most abundant purple nonsulfur bacteria recovered from the blooms, grew in the presence of NaCl concentrations ranging from 0.5 to 3.0% (optimum NaCl concentration, 0.8%) and at pH values ranging from 7.5 to 9.0 (optimum pH, 8.0 to 8.5). On the basis of these unique physiological traits, together with genotypic and phylogenetic data, we propose that the group I isolates should be classified as members of a new species, Rhodovulum strictum. The group II isolates were identified definitely as Rhodovulum sulfidophilum, and the group III and IV isolates were phenotypically most similar to R. sulfidophilum and Rhodovulum adriaticum, respectively, but could be differentiated from these species by DNA-DNA pairing data.


Development and validation of the Liverpool infant bronchiolitis severity score: a research protocol.
To develop and validate a bronchiolitis severity scoring instrument for use by nurses and other healthcare professions. Bronchiolitis is a viral lower respiratory tract infection of infancy. In industrialized countries, admission rates have increased over the last decade with up to 3% of all infants born being admitted to hospital. A small number of these hospitalized infants will require admission to critical care for either invasive or non-invasive ventilation. During the seasonal epidemic, the number of unplanned admissions to critical care with bronchiolitis substantially increases. We will use a mixed methods study design. We will use scale development and psychometric methods to develop a scoring instrument and to test the instrument for content, construct and criterion validity and reliability in several different clinical locations. This study protocol has been reviewed and approved by the NHS National Research Ethics Service, January 2011. There is an urgent need to develop a valid and reliable severity scoring instrument sensitive to clinical changes in the infant, to facilitate clinical decision-making and help standardize patient care. Furthermore, a valid and reliable scoring instrument could also be used as a proxy patient-reported outcome measure to evaluate the efficacy of clinical interventions in randomized controlled trials.


The effect of hypoxia on the chondrogenic differentiation of co-cultured articular chondrocytes and mesenchymal stem cells in scaffolds.
In this work, we investigated the effects of lowered oxygen tension (20% and 5% O2) on the chondrogenesis and hypertrophy of articular chondrocytes (ACs), mesenchymal stem cells (MSCs) and their co-cultures with a 30:70 AC:MSC ratio. Cells were cultured for six weeks within porous scaffolds, and their cellularity, cartilaginous matrix production (collagen II/I expression ratio, hydroxyproline and GAG content) and hypertrophy markers (collagen X expression, ALP activity, calcium accumulation) were analyzed. After two weeks, hypoxic culture conditions had expedited chondrogenesis with all cell types by increasing collagen II/I expression ratio and matrix synthesis by ~2.5-11 and ~1.5-3.0 fold, respectively. At later times, hypoxia decreased cellularity but had little effect on matrix synthesis. ACs and co-cultures showed similarly high collagen II/I expression ratio and GAG rich matrix formation, whereas MSCs produced the least hyaline cartilage-like matrix and obtained a hypertrophic phenotype with eventual calcification. MSC hypertrophy was further emphasized in hypoxic conditions. We conclude that the most promising cell source for cartilage engineering was co-cultures, as they have a potential to decrease the need for primary chondrocyte harvest and expansion while obtaining a stable highly chondrogenic phenotype independent of the oxygen tension in the cultures.


Success rates and complications of awake caudal versus spinal block in preterm infants undergoing inguinal hernia repair: A prospective study.
Inguinal hernia is a common disease in preterm infants necessitating surgical repair. Despite the increased risk of postoperative apnea in preterm infants, the procedure was conventionally performed under general anesthesia. Recently, regional anesthesia approaches, including spinal and caudal blocks have been proposed as safe and efficient alternative anesthesia methods in this group of patients. The current study evaluates awake caudal and spinal blocks in preterm infants undergoing inguinal hernia repair. In a randomized clinical trial, 66 neonates and infants (weight <5 kg) undergoing inguinal hernia repair were recruited in Tabriz Teaching Children Hospital during a 12-month period. They were randomly divided into two equal groups; receiving either caudal block by 1 ml/kg of 0.25% bupivacaine plus 20 μg adrenaline (group C) or spinal block by 1 mg/kg of 0.5% bupivacaine plus 20 μg adrenaline (group S). Vital signs and pain scores were documented during operation and thereafter up to 24 h after operation. Decrease in heart rate and systolic blood pressure was significantly higher in group C throughout the study period (P < 0.05). The mean recovery time was significantly higher in group S (27.3 ± 5.5 min vs. 21.8 ± 9.3 min; P = 0.03). Postoperative need for analgesia was significantly more frequent in group S (75.8% vs. 36.4%; P = 0.001). Failure in anesthesia was significantly higher in group S (24.4% vs. 6.1%; P = 0.04). More appropriate success rate, duration of recovery and postoperative need of analgesics could contribute to caudal block being a superior anesthesia technique compared to spinal anesthesia in awaked preterm infants undergoing inguinal hernia repair.


Adipokine Visfatin's Role in Pathogenesis of Diabesity and Related Metabolic Derangements.
Visfatin is one of the prominent adipokines secreted by adipose tissue. The level of visfatin increases significantly in persons with obesity owing to increased body mass index (BMI). During obesity, the adipocytes, which populate adipose tissue, undergo hypertrophy and hyperplasia and secrete a number of adipocytokines including visfatin. Visfatin, which also acts as an enzyme nicotinamide phosphoribosyl transferase, is one of the prominent adipokines that influence metabolic homeostasis in the body. Visfatin exists in two forms, extracellular and intracellular, and enacts a multitude of actions. The direct and indirect evidence gathered from in-vitro, in-vivo and clinical studies indicate that visfatin modulates obesity and metabolic syndrome-related pathophysiological activities including enhanced inflammation, angiogenesis, synthesis of NAD mononucleotide, and upregulation of antiapoptotic proteins in a number of cell types. It has been implicated in a number of obesity-related alterations and metabolic derangement such as diabetes, cardiovascular complications and some forms of cancers. In this review, the novel hypothesis about the role of visfatin in diabesity has been proposed which implies recent advances in studies about the pathophysiological roles of visfatin during obesity and chronic high glucose in the circulation. Visfatin at high concentration attracts immune cells and produces chronic inflammation in adipocytes. Additionally, it induces insulin resistance in many tissues and causes pancreatic beta cells dysfunction at later stages. Further, its potential as an important target to develop molecular medicine in diabesity and related metabolic syndrome has been highlighted.


Reduced aliquot size for a plasma organochlorine assay for use in epidemiological studies.
Because archived blood specimens are an important but limited resource for conducting epidemiological studies using biomarkers, it is important to develop analytical techniques that minimize the amount of sample needed. We modified an established 1.0-ml blood plasma organochlorine assay to use smaller volumes. We assessed its utility by comparing the accuracy and precision of measurements obtained with different-sized aliquots of spiked plasma from three pools of known concentration of 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) and polychlorinated biphenyls (PCBs; low, medium, and high). There was a modest sacrifice in accuracy using 0.5 as opposed to 1.0 ml. However, the within-batch coefficients of variation, a measure of laboratory error, were consistently low when 0.5-ml aliquots were used. For both DDE and PCB concentrations, this error was less than 5% for the medium and high pools [5-20 parts per billion (ng/ml)] and less than 9% for the low pool (< 1 part per billion). After determining that aliquots of 0.5 ml were sufficient, we performed a blinded quality control analysis of stored plasma. In this study, the within-subject variation was low for DDE and PCBs and substantially lower than the between-subject variation, suggesting that the assay would rank subjects with reasonable precision. Our results suggest that use of 0.5-ml as opposed to 1.0-ml aliquots should not compromise the power of a nested case-control study to detect differences between subjects and would thus save plasma for future research. For populations with very low levels of organochlorines, however, the larger volumes should still be used.


Assessing the palliative care needs and service use of diverse older adults in an urban medically-underserved community.
Although palliative care (PC) has become increasingly familiar, considerable gaps persist in access to and use of services. Community-based programs remain rare, and low-income, minority communities significantly under-utilize hospice and palliative services. We used community-based participatory research (CBPR) methods to conduct a mixed-methods community needs assessment of seriously-ill older adults (n=100) and providers from community-based programs and churches (n=41) in an urban medically-underserved community in the U.S. to explore: (I) the prevalence and severity of illness-related symptoms and psychosocial-spiritual concerns; (II) the scope and quality of community supports helping older adults manage their symptoms; and (III) the perceptions and utilization of palliative and supportive care services among older adults and community-based service providers. Participants reported high rates of chronic illness-related symptoms (i.e., pain, fatigue, sleeping difficulties, depression, and anxiety), and many described unmet needs around symptom management. Few had ever utilized PC or pain management services, and most relied primarily on family, friends, and faith communities to help them manage burdensome symptoms. Barriers included lack of familiarity with PC, limited access and financial concerns. Older adults were largely unfamiliar with PC, and many described unmet needs and desire for help with symptom burden. Findings support the need to further explore community-level and cultural barriers to PC among diverse, underserved older adults. Development of innovative community partnerships may help raise awareness of PC and address the physical and psychosocial-spiritual challenges facing chronically-ill minority older adults and their families.


The addition of continuous intravenous infusion of ketorolac to a patient-controlled analgetic morphine regime reduced postoperative myocardial ischemia in patients undergoing elective total hip or knee arthroplasty.
This double-blind randomized trial assessed the effect of adding an intravenous continuous infusion of ketorolac to a patient-controlled analgesia (PCA) morphine regimen on analgesia, heart rate, arterial blood pressure, and postoperative myocardial ischemia. Patients having elective total hip or knee replacement were randomized to receive ketorolac 30 mg bolus, followed by an infusion of 5 mg/h for 24 h or placebo. All patients had access to PCA morphine (20 microg/kg bolus, with a lockout of 6 min). Patients were monitored for pain visual analog scale, blood pressure, heart rate, and ST segment depression via a continuous Holter monitor. ST depression of 1 mm 60 ms after the J point was considered significant if it lasted more than 1 min. There was no difference in demographics, risk factors, or cardiac medications between the groups. Ketorolac-treated patients had significantly better pain control at 2, 6, and 24 h. There was significant morphine sparing at all times after 3 h. There was no difference in the number of ischemic events between the groups. The ischemic episodes of the patients who received ketorolac occurred at slower heart rates (97 +/- 15 vs 114 +/- 16 bpm, P = 0.001) than those of patients in the placebo group. The duration of ST depression was shorter in ketorolac-treated patients (24 +/- 35 vs 76 +/- 95 min, P < 0.05). All ST depressions were clinically silent. Logistic regression of factors predicting ischemia included the use of calcium channel blockers and low pain score. These results suggest that analgesia with ketorolac reduces the duration of ischemic episodes in the first 24 h postoperatively.


Inner ear and maternal reproductive defects in mice lacking the Hmx3 homeobox gene.
The Hmx homeobox gene family is of ancient origin, being present in species as diverse as Drosophila, sea urchin and mammals. The three members of the murine Hmx family, designated Hmx1, Hmx2 and Hmx3, are expressed in tissues that suggest a common functional role in sensory organ development and pregnancy. Hmx3 is one of the earliest markers for vestibular inner ear development during embryogenesis, and is also upregulated in the myometrium of the uterus during pregnancy. Targeted disruption of the Hmx3 gene results in mice with abnormal circling behavior and severe vestibular defects owing to a depletion of sensory cells in the saccule and utricle, and a complete loss of the horizontal semicircular canal crista, as well as a fusion of the utricle and saccule endolymphatic spaces into a common utriculosaccular cavity. Both the sensory and secretory epithelium of the cochlear duct appear normal in the Hmx3 null animals. The majority of Hmx3 null females have a reproductive defect. Hmx3 null females can be fertilized and their embryos undergo normal preimplantation development, but the embryos fail to implant successfully in the Hmx3 null uterus and subsequently die. Transfer of preimplantation embryos from mutant Hmx3 uterine horns to wild-type pseudopregnant females results in successful pregnancy, indicating a failure of the Hmx3 null uterus to support normal post-implantation pregnancy. Molecular analysis revealed the perturbation of Hmx, Wnt and LIF gene expression in the Hmx3 null uterus. Interestingly, expression of both Hmx1 and Hmx2 is downregulated in the Hmx3 null uterus, suggesting a hierarchical relationship among the three Hmx genes during pregnancy.


Equipment location in hospitals using RFID-based positioning system.
Throughout various complex processes within hospitals, context-aware services and applications can help to improve the quality of care and reduce costs. For example, sensors and radio frequency identification (RFID) technologies for e-health have been deployed to improve the flow of material, equipment, personal, and patient. Bed tracking, patient monitoring, real-time logistic analysis, and critical equipment tracking are famous applications of real-time location systems (RTLS) in hospitals. In fact, existing case studies show that RTLS can improve service quality and safety, and optimize emergency management and time critical processes. In this paper, we propose a robust system for position and orientation determination of equipment. Our system utilizes passive (RFID) technology mounted on flooring plates and several peripherals for sensor data interpretation. The system is implemented and tested through extensive experiments. The results show that our system's average positioning and orientation measurement outperforms existing systems in terms of accuracy. The details of the system as well as the experimental results are presented in this paper.


Intrinsically disordered segments affect protein half-life in the cell and during evolution.
Precise control of protein turnover is essential for cellular homeostasis. The ubiquitin-proteasome system is well established as a major regulator of protein degradation, but an understanding of how inherent structural features influence the lifetimes of proteins is lacking. We report that yeast, mouse, and human proteins with terminal or internal intrinsically disordered segments have significantly shorter half-lives than proteins without these features. The lengths of the disordered segments that affect protein half-life are compatible with the structure of the proteasome. Divergence in terminal and internal disordered segments in yeast proteins originating from gene duplication leads to significantly altered half-life. Many paralogs that are affected by such changes participate in signaling, where altered protein half-life will directly impact cellular processes and function. Thus, natural variation in the length and position of disordered segments may affect protein half-life and could serve as an underappreciated source of genetic variation with important phenotypic consequences.


Increased prevalence of symptoms of rhinitis but not of asthma between 1990 and 2008 in Swedish adults: comparisons of the ECRHS and GA²LEN surveys.
The increase in asthma prevalence until 1990 has been well described. Thereafter, time trends are poorly known, due to the low number of high quality studies. The preferred method for studying time trends in prevalence is repeated surveys of similar populations. This study aimed to compare the prevalence of asthma symptoms and their major determinants, rhinitis and smoking, in Swedish young adults in 1990 and 2008. In 1990 the European Community Respiratory Health Survey (ECRHS) studied respiratory symptoms, asthma, rhinitis and smoking in a population-based sample (86% participation) in Sweden. In 2008 the same symptom questions were included in the Global Allergy and Asthma European Network (GA(2)LEN) survey (60% participation). Smoking questions were however differently worded. The regions (Gothenburg, Uppsala, Umeå) and age interval (20-44 years) surveyed both in 1990 (n = 8,982) and 2008 (n = 9,156) were analysed. The prevalence of any wheeze last 12 months decreased from 20% to 16% (p<0.001), and the prevalence of "asthma-related symptoms" was unchanged at 7%. However, either having asthma attacks or using asthma medications increased from 6% to 8% (p<0.001), and their major risk factor, rhinitis, increased from 22% to 31%. Past and present smoking decreased. From 1990 to 2008 the prevalence of obstructive airway symptoms common in asthma did not increase in Swedish young adults. This supports the few available international findings suggesting the previous upward trend in asthma has recently reached a plateau. The fact that wheeze did not increase despite the significant increment in rhinitis, may at least in part be due to the decrease in smoking.


Influence on photosynthesis of starlight, moonlight, planetlight, and light pollution (reflections on photosynthetically active radiation in the universe).
Photosynthesis on Earth can occur in a diversity of organisms in the photosynthetically active radiation (PAR) range of 10 nmol of photons m(-2) s(-1) to 8 mmol of photons m(-2) s(-1). Similar considerations would probably apply to photosynthetic organisms on Earth-like planets (ELPs) in the continuously habitable zone of other stars. On Earth, starlight PAR is inadequate for photosynthetically supported growth. An increase in starlight even to reach the minimum theoretical levels to allow for photosynthesis would require a universe that was approximately ten million times older, or with a ten million times greater density of stars, than is the case for the present universe. Photosynthesis on an ELP using PAR reflected from a natural satellite with the same size as our Moon, but at the Roche limit, could support a low rate of photosynthesis at full Moon. Photosynthesis on an ELP-like satellite of a Jupiter-sized planet using light reflected from the planet could be almost 1% of the rate in full sunlight on Earth when the planet was full. These potential contributions to photosynthesis require that the contribution is compared with the rate of photosynthesis driven by direct radiation from the star. Light pollution on Earth only energizes photosynthesis by organisms that are very close to the light source. However, effects of light pollution on photosynthesis can be more widespread if the photosynthetic canopy is retained for more of the year, caused by effects on photoperiodism, with implications for the influence of civilizations on photosynthesis.


Ultrafast Dynamics of o-Nitrophenol: An Experimental and Theoretical Study.
The photolysis of o-nitrophenol (o-NP), a typical push-pull molecule, is of current interest in atmospheric chemistry as a possible source of nitrous acid (HONO). To characterize the largely unknown photolysis mechanism, the dynamics of the lowest lying excited singlet state (S1) of o-NP was investigated by means of femtosecond transient absorption spectroscopy in solution, time-resolved photoelectron spectroscopy (TRPES) in the gas phase and quantum chemical calculations. Evidence of the unstable aci-nitro isomer is provided both in the liquid and in the gas phase. Our results indicate that the S1 state displays strong charge transfer character, which triggers excited state proton transfer from the OH to the NO2 group as evidenced by a temporal shift of 20 fs of the onset of the photoelectron spectrum. The proton transfer itself is found to be coupled to an out-of-plane rotation of the newly formed HONO group, finally leading to a conical intersection between S1 and the ground state S0. In solution, return to S0 within 0.2-0.3 ps was monitored by stimulated emission. As a competitive relaxation channel, ultrafast intersystem crossing to the upper triplet manifold on a subpicosecond time scale occurs both in solution and in the gas phase. Due to the ultrafast singlet dynamics, we conclude that the much discussed HONO split-off is likely to take place in the triplet manifold.


Frostbite: Pathophysiology, Epidemiology, Diagnosis, Treatment, and Prevention.
Frostbite can occur during cold-weather operations when the temperature is <0°C (<32°F). When skin temperature is =-4°C (=25°F), ice crystals form in the blood, causing mechanical damage, inflammation, thrombosis, and cellular death. Lower temperatures, higher wind speeds, and moisture exacerbate the process. The frozen part or area should not be rewarmed unless the patient can remain in a warm environment; repeated freeze/thaw cycles cause further injury. Treatment involves rapid rewarming in a warm, circulating water bath 37°C to 39°C (99°F-102°F) or, if this is not possible, then contact with another human body. Thrombolytics show promise in the early treatment of frostbite. In the field, the depth and severity of the injury can be determined with laser Doppler ultrasound devices or thermography. In hospital settings, bone scintigraphy with single-photon emission computed tomography (SPECT) 2 to 4 days postinjury provides detailed information on the depth of the injury. Prevention is focused primarily on covering exposed skin with proper clothing and minimizing exposure to wind and moisture. The Generation III Extended Cold Weather Clothing System is an interchangeable 12-piece clothing ensemble designed for low temperatures and is compatible with other military systems. The Extreme Cold Vapor Barrier Boot has outer and inner layers composed of seamless rubber with wool insulation between, rated for low temperatures. The Generation 3 Modular Glove System consists of 11 different gloves and mitts with design features that assist in enhancing grip, aid in the use of mobile devices, and allow shooting firearms. Besides clothing, physical activity also increases body heat, reducing the risk of frostbite.


Wolbachia-induced parthenogenesis in a genus of phytophagous mites.
The vertically transmitted endosymbiotic bacterium Wolbachia modifies host reproduction in several ways in order to enhance its own spread. One such modification results in the induction of parthenogenesis, where males, which are unable to transmit Wolbachia, are not produced. Interestingly, parthenogenesis-inducing Wolbachia have only been found within haplodiploid insects and it is not known whether this exclusivity is the result of functional constraints of Wolbachia. Here we find a unique pattern of Wolbachia infection that is associated with parthenogenesis in six species within the phytophagous mite genus Bryobia. Through antibiotic treatment we show that, in two species, Bryobia praetiosa and an unidentified species, the Wolbachia infection is strictly associated with parthenogenesis. Microsatellite loci show the mechanism of parthenogenesis to be functionally apomictic and not gamete duplication, with progeny identical to their infected mother. Crossing experiments within B. praetiosa showed no evidence of sexual reproduction. These results are discussed with reference to the distribution of parthenogenesis-inducing Wolbachia and the diversification of the Bryobia genus.


Crystal structure of human antibody 2909 reveals conserved features of quaternary structure-specific antibodies that potently neutralize HIV-1.
Monoclonal antibody 2909 belongs to a class of potently neutralizing antibodies that recognize quaternary epitopes on HIV-1. Some members of this class, such as 2909, are strain specific, while others, such as antibody PG16, are broadly neutralizing; all, however, recognize a region on the gp120 envelope glycoprotein that includes two loops (V2 and V3) and forms appropriately only in the oligomeric HIV-1 spike (gp120(3)/gp41(3)). Here we present the crystal structure of 2909 and report structure-function analysis with antibody chimeras composed of 2909 and other members of this antibody class. The 2909 structure was dominated by a heavy-chain third-complementarity-determining region (CDR H3) of 21 residues, which comprised 36% of the combining surface and formed a β-hairpin club extending ∼20 Å beyond the rest of the antibody. Sequence analysis and mass spectrometry identified sites of tyrosine sulfation at the middle and top of CDR H3; substitutions with phenylalanine either ablated (middle substitution) or substantially diminished (top substitution) neutralization. Chimeric antibodies composed of heavy and light chains, exchanged between 2909 and other members of the class, indicated a substantial lack of complementation. Comparison of 2909 to PG16 (which is tyrosine sulfated and the only other member of the class for which a structure has previously been reported) showed that both utilize protruding, anionic CDR H3s for recognition. Thus, despite some diversity, members of this class share structural and functional similarities, with conserved features of the CDR H3 subdomain likely reflecting prevalent solutions by the human immune system for recognition of a quaternary site of HIV-1 vulnerability.


Saprolegnia strains isolated from river insects and amphipods are broad spectrum pathogens.
Saprolegnia species are destructive pathogens to many aquatic organisms and are found in most parts of the world. Reports based on phylogenetic analysis suggest that Saprolegnia strains isolated from aquatic animals such as crustaceans and frogs are close to Saprolegnia strains isolated from infected fish or fish eggs and vice versa. However, it has often been assumed that host specificity occurs for each individual isolate or strain. Here we demonstrate that Saprolegnia spp. can have multiple hosts and are thus capable of infecting different aquatic organisms. Saprolegnia delica, Saprolegnia hypogyna, and 2 strains of Saprolegnia diclina were isolated from aquatic insects and amphipods while S. delica, Saprolegnia ferax, Pythium pachycaule, and a Pythium sp. were isolated from the water of a medium to fast flowing river. The ITS region of the rRNA gene was sequenced for all isolates. In challenge experiments, all four isolates from insects were found to be highly pathogenic to eggs of Atlantic salmon (Salmo salar) and embryos of the African clawed frog (Xenopus laevis). We found that Saprolegnia spp. isolated from salmon eggs were also able to successfully establish infection in nymphs of stonefly (Perla bipunctata) and embryos of X. laevis). These results suggest that Saprolegnia spp. are capable of infecting multiple hosts, which may give them an advantage during seasonal variation in their natural environments.


Splenic rupture in children with hematologic malignancies.
Splenic rupture is an uncommon but life-threatening complication of leukemias and lymphomas, and is reported mostly in adults. The authors investigated the frequency with which splenic rupture is diagnosed in pediatric patients with hematologic malignancies and reviewed its clinical profile and outcome. The data base of St. Jude Children's Research Hospital was searched for cases coded as splenic laceration or rupture, splenic infarction, or splenectomy in patients diagnosed with lymphoma or leukemia between January 1962 and December 1997. The medical records of patients with histopathologic or radiologic evidence of splenic rupture were reviewed. The time spanned by the study was divided into early (1962-1990) and recent (1991-1997) eras to reflect the availability of modern diagnostic imaging techniques. Seven children experienced splenic rupture. They were between ages 5-17 years. There were four males and three females. Primary diagnoses included acute myeloid leukemia (four patients), acute lymphoblastic leukemia (two patients), and Hodgkin lymphoma (one patient). Five patients were diagnosed in the recent era and two in the early era. Four patients had radiologic or bacteriologic evidence of fungal infection concomitant with the splenic event. Of five deaths, only two were related causally to splenic rupture; these occurred in the early era. All seven acute episodes of splenic rupture were managed conservatively without surgery. The overall frequency with which splenic rupture was detected in children with hematologic malignancy at the study institution was 0.18%. In the recent era, the frequency of detection was 9-fold higher (0.55%) than that of the early era (0.06%). Improved imaging techniques and increased utilization of imaging studies may account for the increased incidental detection of "preclinical" splenic rupture. Adolescent age group, acute myeloid leukemia (especially acute promyelocytic leukemia), a high leukocyte count, thrombocytopenia, and coagulopathy may predispose children with leukemia to pathologic splenic rupture. Fungal infection frequently was associated with splenic rupture and may play a role in its pathogenesis.


Preparation and phase transition of FeOOH nanorods: strain effects on catalytic water oxidation.
The evolution of the phase and morphology of FeOOH nanorods prepared by a hydrothermal method is studied via X-ray diffraction (XRD) and in situ transmission electron microscopy. The FeOOH nanorod with a tetragonal structure (β-FeOOH) is gradually converted into a rhombohedral Fe2O3 nanorod by a simple thermal treatment. The existence of an intermediate FeOOH structure with high lattice strains during the phase transition is identified by Rietveld analysis using XRD. The electrochemical properties of the nanorods are investigated based on the crystal phases to elucidate their relative catalytic activities. The strained-FeOOH nanorods exhibited enhanced catalytic water oxidation activity and stability. Typically, the strained-FeOOH nanorods showed high electrochemical stability under neutral conditions, while tetragonal FeOOH nanorods under the same conditions showed rapid deactivation for water oxidation reaction.


A Historical Review of Diachrony and Semantic Dimensions of Trace in Neurosciences and Lacanian Psychoanalysis.
Experience leaves a trace in the nervous system through plasticity. However, the exact meaning of the mnesic trace is poorly defined in current literature. This article provides a historical review of the term trace in neuroscience and psychoanalysis literature, to highlight two relevant aspects: the diachronic and the semantic dimensions. There has been a general interest in diachrony, or a form of evolution of the trace, but its indissociable semantic dimension remains partially disregarded. Although frequently implied, the diachronic and semantic dimensions of the trace are rarely clearly articulated. We situate this discussion into the classical opposition of syntax, or rules of inscription of the trace in the nervous system, and semantics, or the content of the trace, which takes into consideration the attempt of the human being to build coherence. A general observation is that the study of the term trace follows trends of the thought of the given epoch. This historical analysis also reveals the decay of the idea that the trace is reliable to the experience. From the articulation between neurosciences and psychoanalysis in a historical perspective, this review shows that the trend is to consider trace as a production of the subject, resulting in a permanent rewriting in an attempt to give meaning to the experience. This trend is becoming increasingly evident in light of recent research in neurosciences and psychoanalysis.


Prolonged effect of a novel S-nitrosated glyco-amino acid in endothelium-denuded rat femoral arteries: potential as a slow release nitric oxide donor drug.
1. The vasodilator properties of a novel S-nitrosated glyco-amino acid (RIG200) were investigated in isolated rat femoral arteries and compared with those of the parent S-nitrosothiol compound, S-nitroso-N-acetylpenicillamine (SNAP). 2. Spectrophotometric analysis revealed that 2.5 mM solutions of RIG200 decomposed more slowly (half-life (t1/2) = 216.2 +/- 26.7 min) than SNAP (t1/2 = 37.2 +/- 13.8 min) in Krebs buffer at 24 degrees C. Furthermore, the rate of decomposition of SNAP, but not of RIG200, was significantly reduced by the Cu(I) chelator, neocuproine. We concluded that the relative stability of RIG200 is due, at least in part, to its resistance to trace Cu(I)-catalyzed decomposition. Nitric oxide (NO) generation from SNAP and RIG200 was confirmed by use of an NO electrode. 3. Experiments to investigate the vasodilator effects of RIG200 were carried out on isolated femoral arteries taken from adult male Wistar rats (400-550 g). Lengths of artery (7-8 mm long) were cannulated, dissected free and perfused at constant flow rate (0.6 ml min-1) with Krebs buffer. Vessels were precontracted with phenylephrine (10.2 +/- 0.3 microM) and developed pressures of 91.8 +/- 4 mmHg, detected upstream by a differential pressure transducer. 4. Concentration-dependent vasodilator responses to bolus injections of SNAP or RIG200 (10 microliters; 10(-8)-10(-3) M) made into the perfusate of endothelium-intact vessels were transient, recovering the preinjection pressure in < 20 min. 5. Responses to equivalent bolus injections of SNAP in endothelium-denuded vessels were also transient but those in response to concentrations of RIG200 > 10(-5) M were sustained. Responses to 10(-3) M RIG200 were sustained for periods > 4 h. Sustained vasodilatation was reversed by the NO scavenger, ferrohaemoglobin (10 microM) but was unaffected by the NO synthase inhibitor, N omega-nitro-L-arginine methyl ester (200 microM), indicating involvement of NO from a source other than NO synthase. 6. We suggest that a possible explanation for the prolonged effect of RIG200 is retention of the compound by the vascular wall, facilitated by endothelial denudation. Slow decomposition of RIG200 in situ would release sufficient NO to maintain a 'vasodilator tone' which persists for more than 4 h. Selective retention by damaged vessels could have important therapeutic implications with regard to targeted delivery of NO, restoring protection to areas deprived of endogenous NO, whilst avoiding unwanted hypotension.


3-Nitrotyrosine inhibits fibroblast-mediated collagen gel contraction and chemotaxis.
Reactive nitrogen species induce tissue inflammation and nitrate tyrosine residues of various kinds of proteins. Recent studies have established that the free amino acid form of 3-nitrotyrosine induces cytotoxity and growth inhibition and alters the cellular function in cultured cells. The aim of this study was to evaluate whether 3-nitrotyrosine could affect tissue remodelling in fibroblasts. To accomplish this, human fetal lung fibroblasts (HFL-1) were used to assess the fibroblast-mediated contraction of floating gels and chemotaxis towards fibronectin. In addition, the ability of fibroblasts to release fibronectin, transforming growth factor (TGF)-beta1, fibronectin and vascular endothelial growth factor (VEGF) was assessed. 3-Nitrotyrosine significantly inhibited gel contraction (p<0.01) compared with control and this inhibition was abolished by nitric oxide synthase (NOS) inhibitor. 3-Nitrotyrosine did not affect TGF-beta1 and VEGF but significantly decreased fibronectin release (p<0.01) into the media. 3-Nitrotyrosine significantly inhibited chemotaxis towards fibronectin through suppression of alpha(5)beta(1) integrin expression (p<0.01). NOS inhibitor also reversed 3-nitrotyrosine-inhibited chemotaxis (p<0.01). Finally, 3-nitrotyrosine enhanced the expression of the inducible type of NOS (p<0.01) and nitric oxide release (p<0.01) through nuclear factor-kappaB activation. These results suggest that the free amino acid form of 3-nitrotyrosine can affect the tissue repair process by modulating nitric oxide production.


Cutaneous Notch signaling in health and disease.
The development and maintenance of the skin are dependent on myriad signaling pathways that regulate a variety of cellular processes. In cutaneous epithelial cells, the Notch cascade plays a central role in ensuring that proliferation and differentiation are coordinated appropriately, a function that it imparts during both ontogeny and homeostasis. Aberrations of the Notch signaling pathway result in severe abnormalities in the epidermis and its appendages and cause functional defects such as perturbed barrier function. In addition, impaired Notch signaling is associated with diseases of the skin such as atopy and cancer. The pathology associated with aberrant cutaneous Notch signaling reflects the complex mechanisms underpinning its function in this tissue and involves both cell-autonomous and nonautonomous mechanisms. This review summarizes our current knowledge of the role of Notch signaling in the skin during health and disease.


Primary prevention of lead poisoning in children: a cross-sectional study to evaluate state specific lead-based paint risk reduction laws in preventing lead poisoning in children.
Children younger than 72 months are most at risk of environmental exposure to lead from ingestion through normal mouthing behavior. Young children are more vulnerable to lead poisoning than adults because lead is absorbed more readily in a child's gastrointestinal tract. Our focus in this study was to determine the extent to which state mandated lead laws have helped decrease the number of new cases of elevated blood-lead levels (EBLL) in homes where an index case had been identified. A cross-sectional study was conducted to compare 682 residential addresses, identified between 2000 and 2009, in two states with and one state without laws to prevent childhood lead poisoning among children younger than 72 months, to determine whether the laws were effective in preventing subsequent cases of lead poisoning detected in residential addresses after the identification of an index case. In this study, childhood lead poisoning was defined as the blood lead level (BLL) that would have triggered an environmental investigation in the residence. The two states with lead laws, Massachusetts (MA) and Ohio (OH), had trigger levels of ≥25 μg/dL and ≥15 μg/dL respectively. In Mississippi (MS), the state without legislation, the trigger level was ≥15 μg/dL. The two states with lead laws, MA and OH, were 79% less likely than the one without legislation, MS, to have residential addresses with subsequent lead poisoning cases among children younger than 72 months, adjusted OR = 0.21, 95% CI (0.08-0.54). For the three states studied, the evidence suggests that lead laws such as those studied herein effectively reduced primary exposure to lead among young children living in residential addresses that may have had lead contaminants.


Evidence-Based interventions of Norovirus outbreaks in China.
In resource-limited settings where laboratory capacity is limited and response strategy is non-specific, delayed or inappropriate intervention against outbreaks of Norovirus (NoV) are common. Here we report interventions of two norovirus outbreaks, which highlight the importance of evidence-based modeling and assessment to identify infection sources and formulate effective response strategies. Spatiotemporal scanning, mathematical and random walk modeling predicted the modes of transmission in the two incidents, which were supported by laboratory results and intervention outcomes. Simulation results indicated that contaminated water was 14 to 500 fold more infectious than infected individuals. Asymptomatic individuals were not effective transmitters. School closure for up to a week still could not contain the outbreak unless the duration was extended to 10 or more days. The total attack rates (TARs) for waterborne NoV outbreaks reported in China (n = 3, median = 4.37) were significantly (p < 0.05) lower than worldwide (n = 14, median = 41.34). The low TARs are likely due to the high number of the affected population. We found that school closure alone could not contain Norovirus outbreaks. Overlooked personal hygiene may serve as a hotbed for infectious disease transmission. Our results reveal that evidence-based investigations can facilitate timely interventions of Norovirus transmission.


Senescence gives insights into the morphogenetic evolution of anamniotes.
Senescence represents a mechanism to avoid undesired cell proliferation that plays a role in tumor suppression, wound healing and embryonic development. In order to gain insight on the evolution of senescence, we looked at its presence in developing axolotls (urodele amphibians) and in zebrafish (teleost fish), which are both anamniotes. Our data indicate that cellular senescence is present in various developing structures in axolotls (pronephros, olfactory epithelium of nerve fascicles, lateral organs, gums) and in zebrafish (epithelium of the yolk sac and in the lower part of the gut). Senescence was particularly associated with transient structures (pronephros in axolotls and yolk sac in zebrafish) suggesting that it may play a role in the elimination of these tissues. Our data supports the notion that cellular senescence evolved early in vertebrate evolution to influence embryonic development.


amplimap: a versatile tool to process and analyze targeted NGS data.
amplimap is a command-line tool to automate the processing and analysis of data from targeted next-generation sequencing experiments with PCR-based amplicons or capture-based enrichment systems. From raw sequencing reads, amplimap generates output such as read alignments, annotated variant calls, target coverage statistics and variant allele counts and frequencies for each target base pair. In addition to its focus on user-friendliness and reproducibility, amplimap supports advanced features such as consensus base calling for read families based on unique molecular identifiers and filtering false positive variant calls caused by amplification of off-target loci. amplimap is available as a free Python package under the open-source Apache 2.0 License. Documentation, source code and installation instructions are available at https://github.com/koelling/amplimap.


Molecular epidemiology of hepatitis B virus in Dakar, Sénégal.
Using DNA chip technology and real-time quantitative PCR, molecular profile of HBV strains infecting blood donors and patients in Dakar, Sénégal was studied. All HBsAg-positive blood donors (n = 175) and all patients who presented with chronic hepatitis B (n = 29) between 1st June 2003 and 31st July 2003 were studied. One patient, a blood donor, was coinfected by HCV, and nine patients had anti-HDV antibodies. Few persons in either group were HBeAg-positive. Viral load values were relatively low but correlated with biochemical abnormalities. Patients were infected mainly by genotype E (72%). Patients infected by genotype A (28%) tended to be younger than other patients. There was no significant difference between the blood donors and the patients with hepatitis B as regards virological markers, including viral load, when the HBV genotype was taken into account. The BCP A1762T and G1764A mutations were found in four patients and one patient, respectively; the two mutations were never found in the same patient. The W28* mutation at position 1896 of the core was detected in 19 of the 32 genotyped patients, 18 (83%) of whom had genotype E infection. ALT levels were not influenced by HBV mutations. This study shows a low frequency of clinical signs in HBsAg-positive blood donors, a relatively low level of viral replication, and a high frequency of pre-core mutants in this West African population. These results underline the importance of molecular characterization of HBV infection as specific treatments become available in this region.


Grammatical evolution decision trees for detecting gene-gene interactions.
A fundamental goal of human genetics is the discovery of polymorphisms that predict common, complex diseases. It is hypothesized that complex diseases are due to a myriad of factors including environmental exposures and complex genetic risk models, including gene-gene interactions. Such epistatic models present an important analytical challenge, requiring that methods perform not only statistical modeling, but also variable selection to generate testable genetic model hypotheses. This challenge is amplified by recent advances in genotyping technology, as the number of potential predictor variables is rapidly increasing. Decision trees are a highly successful, easily interpretable data-mining method that are typically optimized with a hierarchical model building approach, which limits their potential to identify interacting effects. To overcome this limitation, we utilize evolutionary computation, specifically grammatical evolution, to build decision trees to detect and model gene-gene interactions. In the current study, we introduce the Grammatical Evolution Decision Trees (GEDT) method and software and evaluate this approach on simulated data representing gene-gene interaction models of a range of effect sizes. We compare the performance of the method to a traditional decision tree algorithm and a random search approach and demonstrate the improved performance of the method to detect purely epistatic interactions. The results of our simulations demonstrate that GEDT has high power to detect even very moderate genetic risk models. GEDT has high power to detect interactions with and without main effects. GEDT, while still in its initial stages of development, is a promising new approach for identifying gene-gene interactions in genetic association studies.


Reliability and validity of the Iranian version of the Pediatric Quality of Life Inventory™ 4.0 Generic Core Scales in adolescents.
The objective of this study was to investigate the reliability and validity of the Iranian version of the Pediatric Quality of Life Inventory™ 4.0 (PedsQL™ 4.0) Generic Core Scales in adolescents After linguistic validation, the Iranian version of the PedsQL™ 4.0 was completed by 848 healthy and 26 chronically ill adolescents aged 13-18 years and their parents. The internal consistency as measured by Cronbach's alpha coefficients exceeded the minimum reliability standard of .70. No floor effects were observed. Ceiling effects detected ranged from 1.5% for adolescent self-report total scale score to 42.2% for self-report social functioning. All monotrait-multimethod correlations were higher than multitrait-multimethod correlations. The intraclass correlation coefficients (ICC) between adolescent self-report and parent proxy-report showed good to excellent agreement. Exploratory factor analysis supported mainly comparable results with the original US English dialect version. The results of the confirmatory factor analysis for 5-factor models for both self-report and proxy-report indicated acceptable fit for the proposed models. Regarding gender and health status, as hypothesized from previous studies, girls reported lower health-related quality of life than boys on the total score, physical and emotional functioning, and healthy adolescents reported significantly higher health-related quality of life than those with chronic illnesses. The findings support the initial reliability and validity of the Iranian version of the PedsQL™ 4.0 as a generic instrument to measure HRQOL of adolescents in Iran.


Algorithm for ion beam figuring of low-gradient mirrors.
Ion beam figuring technology for low-gradient mirrors is discussed. Ion beam figuring is a noncontact machining technique in which a beam of high-energy ions is directed toward a target workpiece to remove material in a predetermined and controlled fashion. Owing to this noncontact mode of material removal, problems associated with tool wear and edge effects, which are common in conventional contact polishing processes, are avoided. Based on the Bayesian principle, an iterative dwell time algorithm for planar mirrors is deduced from the computer-controlled optical surfacing (CCOS) principle. With the properties of the removal function, the shaping process of low-gradient mirrors can be approximated by the linear model for planar mirrors. With these discussions, the error surface figuring technology for low-gradient mirrors with a linear path is set up. With the near-Gaussian property of the removal function, the figuring process with a spiral path can be described by the conventional linear CCOS principle, and a Bayesian-based iterative algorithm can be used to deconvolute the dwell time. Moreover, the selection criterion of the spiral parameter is given. Ion beam figuring technology with a spiral scan path based on these methods can be used to figure mirrors with non-axis-symmetrical errors. Experiments on SiC chemical vapor deposition planar and Zerodur paraboloid samples are made, and the final surface errors are all below 1/100 lambda.


Goose management schemes to resolve conflicts with agriculture: Theory, practice and effects.
In 2012, the four countries hosting the Svalbard population of pink-footed goose Anser brachyrhynchus along its flyway launched an International Species Management Plan for the population. One of the aims was to reduce conflicts between geese and agriculture to an acceptable level. Since 2006, Norway has offered subsidies to farmers that provide refuge areas for geese on their land. We evaluate the mid-Norwegian goose management subsidy scheme, with a view to its adjustment to prevailing ecological and socio-economic parameters. The analysis indicates that the legitimacy of the scheme is highly dependent on transparency of knowledge management and accountability of management scheme to the farming community. Among farmers, as well as front-line officials, outcomes of prioritisation processes within the scheme are judged unfair when there is an evident mismatch between payments and genuine damage. We suggest how the scheme can be made more fair and responsive to ecological changes, within a framework of adaptive management.


Dynamic Regional Brain Atrophy Rates in the First Year After Ischemic Stroke.
Brain atrophy can be regarded as an end-organ effect of cumulative cardiovascular risk factors. Accelerated brain atrophy is described following ischemic stroke, but it is not known whether atrophy rates vary over the poststroke period. Examining rates of brain atrophy allows the identification of potential therapeutic windows for interventions to prevent poststroke brain atrophy. We charted total and regional brain volume and cortical thickness trajectories, comparing atrophy rates over 2 time periods in the first year after ischemic stroke: within 3 months (early period) and between 3 and 12 months (later period). Patients with first-ever or recurrent ischemic stroke were recruited from 3 Melbourne hospitals at 1 of 2 poststroke time points: within 6 weeks (baseline) or 3 months. Whole-brain 3T magnetic resonance imaging was performed at 3 time points: baseline, 3 months, and 12 months. Eighty-six stroke participants completed testing at baseline; 125 at 3 months (76 baseline follow-up plus 49 delayed recruitment); and 113 participants at 12 months. Their data were compared with 40 healthy control participants with identical testing. We examined 5 brain measures: hippocampal volume, thalamic volume, total brain and hemispheric brain volume, and cortical thickness. We tested whether brain atrophy rates differed between time points and groups. A linear mixed-effect model was used to compare brain structural changes, including age, sex, years of education, a composite cerebrovascular risk factor score, and total intracranial volume as covariates. Atrophy rates were greater in stroke than control participants. Ipsilesional hemispheric, hippocampal, and thalamic atrophy rates were 2 to 4 times greater in the early versus later period. Regional atrophy rates vary over the first year after stroke. Rapid brain volume loss in the first 3 months after stroke may represent a potential window for intervention. Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT02205424.


Atrial natriuretic factor receptor subtypes in the rat central nervous system.
In this study we investigated the presence and anatomical location of atrial natriuretic factor (ANF) receptor subtypes in the rat central nervous system using in vitro autoradiographic and cross-linking techniques. 125I-ANF-(Ser99-Tyr126) served as a labeled ligand, whereas ANF-(Ser99-Tyr126) and two peptides endowed with selectivity for ANF-C receptor--namely, C-ANF (des-[Gln116-Gly120] ANF-[Asp102-Cys121]-NH2) and ANF-(Phe106-Ile113)-NH2--were used as displacing agents. Distribution studies revealed the presence of specific ANF binding sites in a number of central nervous system areas examined. C-ANF at 10(-6) M competed for 125I-ANF binding to a much lower extent than ANF in many of those structures, whereas ANF-(106-113)-NH2 at 10(-6) M did not have a significant effect on the radioligand binding except in the choroid plexus, pia-arachnoid, and olfactory bulb. Analysis of the competition curves revealed that in the choroid plexus, pia-arachnoid, olfactory bulb, subfornical organ, area postrema, and habenular nucleus, ANF interacts with its binding sites with high affinity (IC50, 0.46-0.77 nM). In contrast, C-ANF and ANF-(106-113)-NH2 competed for 125I-ANF binding with high potency (IC50, 2-16 nM) in the choroid plexus and pia-arachnoid only, where they were able to displace 60-70% of the radioligand binding. 125I-ANF cross-linking to olfactory bulb membranes resolved a specific 120-kDa band corresponding to the high molecular weight receptor but did not disclose a specifically labeled band corresponding to the low molecular mass receptor.(ABSTRACT TRUNCATED AT 250 WORDS)


The Influences of Cell Type and ZnO Nanoparticle Size on Immune Cell Cytotoxicity and Cytokine Induction.
Nanotechnology represents a new and enabling platform that promises to provide a range of innovative technologies for biological applications. ZnO nanoparticles of controlled size were synthesized, and their cytotoxicity toward different human immune cells evaluated. A differential cytotoxic response between human immune cell subsets was observed, with lymphocytes being the most resistant and monocytes being the most susceptible to ZnO nanoparticle-induced toxicity. Significant differences were also observed between previously activated memory lymphocytes and naive lymphocytes, indicating a relationship between cell-cycle potential and nanoparticle susceptibility. Mechanisms of toxicity involve the generation of reactive oxygen species, with monocytes displaying the highest levels, and the degree of cytotoxicity dependent on the extent of nanoparticle interactions with cellular membranes. An inverse relationship between nanoparticle size and cytotoxicity, as well as nanoparticle size and reactive oxygen species production was observed. In addition, ZnO nanoparticles induce the production of the proinflammatory cytokines, IFN-γ, TNF-α, and IL-12, at concentrations below those causing appreciable cell death. Collectively, these results underscore the need for careful evaluation of ZnO nanoparticle effects across a spectrum of relevant cell types when considering their use for potential new nanotechnology-based biological applications.


Psychometric properties and prognostic usefulness of the Youth Psychopathic Traits Inventory (YPI) as a component of a clinical protocol for detained youth: A multiethnic examination.
Prior studies have shown that the Youth Psychopathic Traits Inventory (YPI) holds promise as a self-report tool for assessing psychopathic traits in detained adolescents. However, these studies have been conducted in a research context where anonymity and confidentiality are provided. Few studies have examined the usefulness of the YPI in clinical settings. To address this research gap, the present study examined data from 1,559 detained boys who completed the YPI as part of a clinical protocol. Official criminal records were available for a subsample (n = 848), allowing us to test the prognostic usefulness of the YPI. Results of confirmatory factor analyses, overall, support the proposed 3-factor structure, though model fit indices were not as good in Dutch boys compared to boys from other ethnic groups. Measurement invariance tests showed that the YPI scores are manifested in the same way across all 4 ethnic groups and suggest that means scores between the 4 ethnic groups are comparable. The YPI scores were internally consistent, and correlations with external variables, including aggression and conduct problems, support the convergent validity of the interpretation of YPI scores. Finally, results demonstrated that YPI scores were not significantly positively related to future criminality. In conclusion, this study suggests that the YPI may hold promise as a self-report tool for assessing psychopathic traits in detained male adolescents during a clinical protocol. However, the finding that the YPI did not predict future offending suggests that this tool should not yet be used for risk assessment purposes in forensic settings. (PsycINFO Database Record


Novel multiple endocrine neoplasia type 1 variations in patients with sporadic primary hyperparathyroidism.
Primary hyperparathyroidism (PHPT) can occur either as a sporadic case or in association with syndromes such as multiple endocrine neoplasia. Multiple endocrine neoplasia type 1 (MEN1) is a rare autosomal-dominant disease resulting from mutations in MEN1 gene encoding a 621 amino acid long tumor suppressor protein "menin." We report here the results of MEN1 screening in 31 patients diagnosed with sporadic PHPT. Diagnosis of sporadic PHPT was made when blood urea and serum creatinine were normal, serum parathyroid hormone was high, and parathyroid enlargement could be localized on ultrasound and/or parathyroid scan. A total of 31 patients and 50 healthy volunteers were recruited for molecular analysis after taking informed consent. Major symptoms at presentation were bone pain, fatigue, muscle weakness, and renal stones. Molecular genetic analysis revealed the presence of two novel intronic variations, c. 913-79T>A and c. 784-129T>A which by human splicing finder are predicted to cause potential alteration of splicing by either activating an intronic cryptic acceptor site or converting a conserved exonic splicing silencer sequence to an exonic splicing enhancer site. Apart from these, two reported polymorphisms rs144677807 and rs669976 were seen only in patients and none of the controls. Other reported polymorphisms rs2071313 and rs654440 were identified both in controls and patients. This is the first study of MEN1 gene screening in sporadic PHPT in India reporting on the clinical and genetic findings, wherein two novel intronic variations c. 913-79T>A and c. 784-129T>A were identified showing their possible role in disease causation.


Evaluation of the current status of the eye lens radiation exposure in an Interventional Radiology department.
Following recent epidemiological studies, which showed tissue reactions from ionizing radiation at significantly lower doses, the 2013/59 EURATOM Directive of 5th December 2013 lowered the limit on the equivalent dose to the eye lens from 150 mSv to 20 mSv per year. Therefore, as a precautionary measure, it is considered appropriate to perform a timely dose monitoring by using specific dosimeters. Analysis of the current state of the eye lens exposures during interventional procedures. The survey aimed at assessing the degree of information available to the exposed workers as regards lowering the dose limit in Interventional Radiology departments of some hospitals in Campania (Southern Italy). The equivalent dose was assessed, over a period of 90 days, using specific Hp dosimeters(3), placed sideways with regard to prescription eye glasses. The level of awareness of the new dose limit among operators was assessed using a questionnaire. The values of the equivalent dose to the lens of the eye for the I and II Operators were found to be <150 mSv/year but for the I Operator a value of 54 mSv/year was obtained, ie higher than 20 mSv/year, that is the new limit of the equivalent dose according to 2013/59 EURATOM. The initial results of the questionnaire from 52 exposed workers, of which 46 (88%) were from exposure category A and 6 (12%) from category B, showed a low level of information (19%). The results highlight not only the importance of using specific devices for individual protection but also the importance of the level of training and information the exposed medical staff are given concerning the new regulations.


HIV infection and patterns of risk among women drug injectors and crack users in low and high sero-prevalence sites.
As AIDS cases among US women continue to increase, a better understanding of women's behavioural risk patterns is needed to inform intervention efforts. Data were from 2,945 women drug injectors and crack users. Statistical analyses compared sociodemographic variables, lifetime behavioural risk patterns, HIV sero-prevalence and history of sexually transmitted diseases, and determined predictors of HIV infection separately in 16 low and four high sero-prevalence sites. Based on risk patterns, four behaviourally-defined sub-groups were constructed, and rates of HIV sero-prevalence were compared. In comparisons between low and high sero-prevalence sites, there were significant differences on most variables examined, and in the relative importance of the sociodemographic characteristics and risk patterns predicting HIV. Drug injection and sex exchange were each independent, significant, behavioural predictors of infection, with no significant difference between the odds ratios attributed to each predictor. HIV sero-prevalence was significantly different among four sub-groups. Interventions must be tailored to address observed differences among women in low and high sero-prevalence sites. Injection drug use and exchanging sex each play a major role in the transmission of HIV infection to US women. Prevention efforts targeted at women should address differences in behavioural risk patterns. Aggressive and innovative interventions are needed for women who exchange sex. AIDS research must investigate how socioeconomic factors impact women's risk for HIV infection.


High phosphorus level leads to aortic calcification via β-catenin in chronic kidney disease.
Vascular calcification is a risk factor for causing cardiovascular events and has a high prevalence among chronic kidney disease (CKD) patients. However, the molecular mechanism underlying this pathogenic process is still obscure. Vascular smooth muscle cells (VSMCs) were induced by a concentration of phosphorus (Pi) of 2.5 mM, and were subjected to cell calcification analyses. The effect of high Pi on the Wnt/β-catenin pathway was measured using a TOP/FOP-Flash reporter assay. The transcriptional regulation of β-catenin on PIT1 (a type III sodium-dependent phosphate cotransporter) was confirmed by promoter reporter and chromatin immunoprecipitation assays. The 5/6 nephrectomized rat was used as an in vivo model and was fed a high Pi diet to induce aortic calcification. Serum levels of phosphate, calcium, creatine, and blood urea nitrogen were measured, and abdominal aortic calcification was examined. High Pi induced VSMC calcification, downregulated expression levels of VSMC markers, and upregulated levels of osteogenic markers. High Pi activated the Wnt/β-catenin pathway and β-catenin activity. β-Catenin was involved in the process of high Pi-induced VSMC calcification. Further investigation revealed that β-catenin transcriptionally regulated Pit1, a necessary player in VSMC osteogenic phenotype change and calcification. The in vivo study showed that β-catenin was involved in rat abdominal aortic calcification induced by high Pi. When knockdown expression of β-catenin in the rat model was investigated, we found that aortic calcification was reduced. These results suggest that β-catenin is an important player in high phosphorus level-induced aortic calcification in CKD.


Dyed-polyvinyl alcohol films: molecular weight and hydrolysis degree influence on optical recording.
An analysis of different polyvinyl alcohol films dyed with Malachite Green is presented. Absorbance and diffraction efficiency of holographic gratings are compared, taking as a parameter the molecular weight and hydrolysis degree of the polymer. It is observed that, using the same dye concentration, the absorption coefficient of the films increases as the molecular weight increases. The absorbance of these plates can be modified when exposed to UV light. In addition, it is found that for holographic recording there is an optimal dye-polymer system film.


Management of pediatric allergic reaction: Practice patterns of Canadian pediatric emergency physicians.
Anaphylaxis is a severe allergic reaction that can be life-threatening. The literature indicates that the incidence of anaphylaxis is increasing and that there are deficiencies in both recognition and management. We aimed to examine the magnitude of these gaps in Canadian pediatric emergency medicine (PEM). We conducted a self-administered survey of the Pediatric Emergency Research Canada (PERC) physician database. The survey tool was developed through a literature review to identify recurring themes of gaps in anaphylaxis diagnosis and management. The final tool contained four scenarios; three scenarios featured each of the National Institute of Allergy and Infectious Diseases (NIAID) anaphylaxis criteria, separately, and a fourth case of non-anaphylactic allergy. Multiple-choice questions associated with each scenario addressed diagnosis, management, and disposition. Additional questions focused on epinephrine prescribing, observation durations, and respondent demographics. Of the 214 members invited to participate in the survey, 152 (71%) responded. Anaphylaxis was accurately recognized 93%, 82%, and 99% of the time for the NIAID criteria one through three, respectively. When anaphylaxis was recognized, epinephrine was prescribed for each case 96%, 95%, and 72% of the time, respectively. Of all respondents, 115 (76%) accurately diagnosed all three cases of anaphylaxis and 82 (54%) treated anaphylaxis with epinephrine each time it was indicated. Most respondents recognized cases of anaphylaxis; however, a substantial number demonstrated gaps in management that may adversely impact this vulnerable population. The recognition of anaphylaxis without urticaria or pulmonary findings and treatment of anaphylaxis with epinephrine, where indicated, were the main gaps identified.


Differentiation of the sarcoplasmic reticulum and T system in developing chick skeletal muscle in vitro.
The electron microscope was used to investigate the first 10 days of differentiation of the SR and the T system in skeletal muscle cultured from the breast muscle of 11-day chick embryos. The T-system tubules could be clearly distinguished from the SR in developing muscle cells fixed with glutaraldehyde and osmium tetroxide. Ferritin diffusion confirmed this finding: the ferritin particles were found only in the tubules identified as T system. The proliferation of both membranous systems seemed to start almost simultaneously at the earliest myotube stage. Observations suggested that the new SR membranes developed from the rough-surfaced ER as tubular projections. The SR tubules connected with one another to form a network around the myofibril. The T-system tubules were formed by invagination of the sarcolemma. The early extension of the T system by branching and budding was seen only in subsarcolemmal regions. Subsequently the T-system tubules could be seen deep within the muscle cells. Immediately after invaginating, the T-system tubule formed, along its course, specialized connections with the SR or ER: triadic structures showing various degrees of differentiation. The simultaneous occurrence of myofibril formation and membrane proliferation is considered to be important in understanding the coordinated events resulting in the differentiated myotube.


Nanocapillary electrokinetic tracking for monitoring charge fluctuations on a single nanoparticle.
We introduce nanoCapillary Electrokinetic Tracking (nanoCET), an optofluidic platform for continuously measuring the electrophoretic mobility of a single colloidal nanoparticle or macromolecule in vitro with millisecond time resolution and high charge sensitivity. This platform is based on using a nanocapillary optical fiber in which liquids may flow inside a channel embedded inside the light-guiding core and nanoparticles are tracked using elastic light scattering. Using this platform we have experimentally measured the electrophoretic mobility of 60 nm gold nanoparticles in an aqueous environment. Further, using numerical simulations, we demonstrate the underlying electrokinetic dynamics inside the nanocapillary and the necessary steps for extending this method to probing single biomolecules, which can be achieved with existing technologies. This achievement will immensely facilitate the daunting challenge of monitoring biochemical or catalytic reactions on a single entity over a wide range of timescales. The unique measurement capabilities of this platform pave the way for a wide range of discoveries in colloid science, analytical biochemistry, and medical diagnostics.