Environmental lead contamination in Miami inner-city area.
The purpose of the study was to evaluate the magnitude of environmental lead contamination in the downtown area of Miami. Lead inspections took place at 121 homes in Little Haiti and Liberty City and involved the collection of representative samples from floors, window wells, tap water, soil and air. Community health workers (CHWs) trained in interview and safety techniques went from door to door to enlist participation. On-site investigations were tailored to areas most utilized by children under the age of 6 years. The presence of lead-containing paint was also investigated in situ via X-ray fluorescence (XRF) analysis. Of the sampling areas, the window wells area had the most abundant occurrence of lead. On analysis, 24% of sites returned window well samples with lead levels above Department of Housing and Urban Development (HUD) guidelines. Of the soil samples, the playgrounds around the house had the highest concentration of lead. Soil sampling demonstrated that 27.5% of sites returned samples with lead levels (400 to 1600 ppm) in excess of HUD/Environmental Protection Agency (EPA) standards. Positive XRF readings in one or more components were returned by 18% of sites. More than half of the houses in these two neighborhoods exhibited unacceptably high levels of lead dust and soil in areas where children live and play. Limitations of this study did not allow the assessment of how many children in this area are affected. A more comprehensive study including other areas of Miami-Dade County with older housing stock is recommended.


[Isolation, purification and properties of restriction endonuclease EcoRII].
The restriction endonuclease Eco RII was isolated and purified to homogeneity. The isolation procedure involved the use of the E. coli strain B834/pSK323, containing the recombinant plasmide pSK323 which provides for the oversynthesis of Eco RII enzymes. Data from gel filtration and Na-DS electrophoresis suggest that the restriction endonuclease Eco RII is a protein made up of two subunits, each with molecular weight of 44 000.


Multiple emulsions controlled by stimuli-responsive polymers.
The phase inversion of water-toluene emulsions stabilized with a single thermo- and pH-sensitive copolymer occurs through the formation of multiple emulsions. At low pH and ambient temperature, oil in water emulsions are formed which transform into highly stable multiple emulsions at pHs immediately lower than the inversion border. At higher pHs, the emulsion turns into a water in oil one.


Dynein light intermediate chain: an essential subunit that contributes to spindle checkpoint inactivation.
The dynein light intermediate chain (LIC) is a subunit unique to the cytoplasmic form of dynein, but how it contributes to dynein function is not fully understood. Previous work has established that the LIC homodimer binds directly to the dynein heavy chain and may mediate the attachment of dynein to centrosomes and other cargoes. Here, we report our characterization of the LIC in Drosophila. Unlike vertebrates, in which two Lic genes encode multiple subunit isoforms, the Drosophila LIC is encoded by a single gene. We determined that the single LIC polypeptide is phosphorylated, and that different phosphoisoforms can assemble into the dynein motor complex. Our mutational analyses demonstrate that, similar to other dynein subunits, the Drosophila LIC is required for zygotic development, germline specification of the oocyte, and mitotic cell division. We show that RNA interference depletion of LIC in Drosophila S2 cells does not block the recruitment of a dynein complex to kinetochores, but it does delay inactivation of Mad2 signaling and mitotic progression. Our observations suggest the LIC contributes to a broad range of dynein functions.


Method for Remote Determination of Object Coordinates in Space Based on Exact Analytical Solution of Hyperbolic Equations.
Accurate remote determination of the object coordinates in 3D space is one of the main questions in many applications. In one of the most popular methods, such determination of the location of an object uses the measurement by receiving an electromagnetic signal transmitted by several spatially distributed base stations (BS). The main problem is that it is necessary to reduce errors and computation time. To overcome these difficulties, an analytical method for determining the position of an object based on the analysis of time difference of arrival (TDoA) of signals from the transmitter of the object to the receivers of the BS is proposed. One of the main advantages of this method is that it is possible to eliminate the ambiguity in determining the coordinates of the object in space and to increase the accuracy of determining the coordinates when the TDoA measurement between base stations fluctuates. Applications for autonomous automotive vehicles and spacebased positioning systems are analyzed. The results obtained show that the proposed algorithm has an accuracy of determining coordinates several times higher than the method of linearization of hyperbolic equations and is less sensitive to TDoA fluctuations at base stations.


Prevalence of anaemia among adolescent girls in the urban slums of Vellore, south India.
A community-based, cross-sectional study was conducted to determine the prevalence of anaemia among unmarried, adolescent south Indian girls in an urban slum setting. A total of 100 apparently healthy girls between the ages of 11 and 18 years were recruited. Their socioeconomic, dietary and anthropometric information was collected, and blood haemoglobin (Hb) was estimated. The prevalence of anaemia (Hb < 12 g%) was 29%. Most had mild anaemia; severe anaemia was not seen. Two-thirds of those with anaemia had low serum ferritin (<12 microg/L). Significant associations were observed between anaemia and low socioeconomic status, religion and reporting infrequent/non-consumption of meat (heme iron). Only meat consumption was related to haemoglobin by multiple regression analysis. Anaemia is a common problem among adolescent girls in this setting, though severe anaemia is rare. There is a need to improve their haemoglobin status through dietary modification along with preventive supplementation and nutrition education.


Compartment resolved reference proteome map from highly purified naïve, activated, effector, and memory CD8⁺ murine immune cells.
Differentiation of CD8(+) T lymphocytes into effector and memory cells is key for an adequate immune response and relies on complex interplay of pathways that convey signals from the cell surface to the nucleus. In this study, we investigated the proteome of four cytotoxic T-cell subtypes; naïve, recently activated effector, effector, and memory cells. Cells were fractionated into membrane, cytosol, soluble nuclear, chromatin-bound, and cytoskeletal compartments. Following LC-MS/MS analysis, identified peptides were analyzed via MaxQuant. Compartment fractionation and gel-LC-MS separation resulted in 2399 proteins identified in total. Comparison between the different subsets resulted in 146 significantly regulated proteins for naïve and effector cells, followed by 116 for activated, and 55 for memory cells. Besides Granzyme B signaling (for activated and/ or effector cells vs. naïve cells), the most prominent changes occurred in the TCA cycle and aspartate degradation. These changes suggest that correct balancing of metabolism is key for differentiation processes. All MS data have been deposited in the ProteomeXchange with identifier PXD001065 (http://proteomecentral.proteomexchange.org/dataset/PXD001065).


Bilateral labyrinthectomy causes long-term deficit in object recognition in rat.
It has been reported that patients with vestibular disorders experience a wide range of cognitive disorders, including memory loss. However, to our knowledge, no study has investigated the contribution of vestibular information to episodic memory in experimental animals using vestibular deafferentation. In the present study, the effects of a complete unilateral or bilateral surgical lesion of the vestibular labyrinths in a spontaneous object recognition task were evaluated in Wistar rats 3 and 6 months following the surgery. We found that rats with bilateral vestibular deafferentation, but not those with unilateral vestibular deafferentation were impaired on the task at both time points. These results suggest for the first time that vestibular information may contribute to non-spatial memory to some extent.


Transcellular and paracellular elements of salt chemosensation in toad skin.
Dehydrated toads absorb water by pressing a specialized (seat patch) area of the skin to moist surfaces. This behavior, the water absorption response (WR), is preceded by periods of more limited skin contact (seat patch down, SPD) in which the suitability of the rehydration source is evaluated. WR and SPD behaviors were suppressed on 250 mM NaCl and 200 mM KCl solutions. Ten micromolar amiloride partially restored SPD and WR on NaCl solutions. The addition of 5 mM La(3+) also partially restored the initiation of WR and this effect was additive to the effect of amiloride, suggesting transcellular and paracellular pathways exist in parallel. Similarly, 5 mM La(3+) partially restored the initiation of WR on KCl solutions, to levels comparable to those with K(+)gluconate, suggesting a paracellular pathway for detection of K(+). Hyperosmotic (250 mM) NaCl solutions bathing the mucosal surface rapidly and reversibly increased the paracellular conductance of isolated skin and this increase was partially inhibited by 5 mM La(3+). These results suggest that the regulation of tight junctions has a chemosensory role in toad skin.


[Polymorphism of glutathion-S-transferase genes in patients with prostatic cancer].
The study of polymorphic variants of GSTT1, GSTM1 and GSTP1 genes from 61 patients with prostatic cancer (PC) has shown that incidence of 0/0 genotype GSTT1 and GSTM1 in PC patients was significantly higher of that in healthy men (n = 100) (34.4 and 15% in p = 0.007 and 60.7 and 43% in p = 0.04, respectively). PC risk in carriers of a GSTT1 deletion form was 2.97, CI95%--1.3-6.84, GSTM1--2.04 in CI95% 1.02-4.1. The analysis of combinations of pathological genotypes of xenobiotic biotransformation enzymes has demonstrated that 89.8% PC patients have a mutation in one of the genes GSTT1, GSTM1 or GSTP1.


Breast cancer prognosis in a mixed Caucasian-Hispanic population.
Prognostic factors were compared between 1,249 Caucasian and 360 Hispanic women with breast cancer. A significantly greater proportion of Hispanic women were less than 50 years of age at diagnosis compared to Caucasian women (P less than .0001). Significantly more Hispanic women presented with tumors larger than 3 cm in diameter and with positive axillary lymph nodes than did Caucasian women (P = .004 and P = .0001, respectively). Significantly more Hispanic women were estrogen receptor (ER) negative (P = .005). However, when examined by age groups, the relationships between ethnicity and extent of disease and ER status were observed only among women over 50 years of age. Multivariate analyses demonstrated that there was no difference in survival between Caucasian and Hispanic women once adjustments were made for the number of positive lymph nodes and ER status. Although complete data were not available, it appeared that the incidence of breast cancer is lower in this population of Hispanic women than in Caucasian women.


High resolution typing of Chlamydophila psittaci by multilocus VNTR analysis (MLVA).
A multilocus VNTR analysis (MLVA) system for detection of tandem repeats across the whole genome of Chlamydophila psittaci has been developed. Twenty selected genetic loci were initially tested on 9 avian reference strains including representatives of all major serotypes (A to F). Thereafter, 8 loci were retained for a more complete study performed on over 150 C. psittaci isolates from different bird species and geographical origins. Comparative analysis of the MLVA results and those obtained from currently available methods including serotyping and/or ompA sequencing indicate that the MLVA system provides an additional level of discrimination, with 20 distinct patterns identified to date. The newly developed MLVA system therefore provides a highly sensitive, high resolution test for the differentiation of C. psittaci isolates from different origins that is suitable for molecular epidemiological studies.


Patterned colloidal deposition controlled by electrostatic and capillary forces.
We use substrates chemically micropatterned with anionic and cationic regions to govern the deposition of charged colloidal particles. The direct observation of the colloidal assembly suggests that this process includes two steps: an initial patterned attachment of colloids to the substrate and an additional ordering of the structure upon drying. The driving forces of the process, i.e. , screened electrostatic and lateral capillary interactions, are discussed. This approach makes it possible to fabricate complex, high-resolution two-dimensional arrays of colloidal particles.


Assessing Frailty in Patients Undergoing Destination Therapy Left Ventricular Assist Device: Observations from Interagency Registry for Mechanically Assisted Circulatory Support.
Frailty and heart failure share common pathways with symptoms that often coexist. Assessment of frailty may inform patient selection for left ventricular assist device (LVAD) therapy. Using Interagency Registry for Mechanically Assisted Circulatory Support (INTERMACS) data of destination therapy (DT) LVAD patients from January 1, 2012, to March 31, 2014, we examined preimplantation provider-assessed frailty and gait speed testing and the association with 1 year postimplantation outcomes. Of 2,469 patients, 227 (9.2%) had provider-assessed frailty. Only 320 (13.0%) completed gait speed testing, whereas 1,047 (42.4%) were "too sick" to perform the test. Provider-assessed frail and nonfrail patients had similar distributions of INTERMACS profiles and similar median gait speeds. One year mortality was higher for patients with provider-assessed frailty versus nonfrail (24.6% vs. 18.9%; p = 0.01) and for those too sick to complete gait speed testing versus completed testing (22.0% vs. 15.9%). There was an association between provider-assessed frailty and mortality, although it was not clinically significant after adjustment (hazard ratio [HR]: 1.38 [95% confidence interval {CI}: 0.97-1.95]). Useful information regarding frailty on postimplant mortality is gained from provider assessment of frailty or knowing gait speed could not be performed. Development of frailty measures better suited for DT LVAD candidates may help in distinguishing between a frailty phenotype and a more reversible from heart failure-related vulnerability.


Differential role of ethanol and acetaldehyde in the induction of oxidative stress in HEP G2 cells: effect on transcription factors AP-1 and NF-kappaB.
The oxidative metabolism of ethanol by the cytochrome P450 2E1 (CYP2E1) has been recognized to contribute to the ethanol-induced deleterious effects through the induction of oxidative stress. This study compared the effect of ethanol and acetaldehyde in the induction of oxidative stress and activation of transcription factors nuclear factor-kappaB (NF-kappaB) and activating protein 1 (AP-1) in HepG2 cells, which do not express CYP2E1, and HepG2 cells transfected with CYP2E1 (E47 cells). Neither ethanol (80 mmol/L) nor acetaldehyde (25-200 micromol/L) caused oxidative stress in HepG2 cells, an effect that was independent of blocking reduced glutathione (GSH) synthesis with buthionine-L-sulfoximine (BSO). However, BSO preincubation caused an overproduction of peroxides and activation of NF-kappaB and AP-1 in E47 cells even in the absence of ethanol. Furthermore, the incubation of E47 cells with ethanol (80 mmol/L for up to 5 days) depleted cellular GSH stores in both cytosol and mitochondria, reflecting the induction of oxidative stress. Ethanol activated NF-kappaB and AP-1 in E47 cells, an effect that was prevented by 4-methylpyrazole, potentiated by cyanamide, and attenuated by trolox C. Interestingly, however, despite the inability of acetaldehyde to induce oxidative stress in HepG2, acetaldehyde activated NF-kappaB and AP-1; in contrast, ethanol failed to activate these transcription factors in HepG2. Thus, our findings indicate that activation of NF-kappaB and AP-1 by ethanol and acetaldehyde occurs through distinct mechanisms. CYP2E1 is indispensable in the induction of oxidative stress from ethanol, whereas the activation of NF-kappaB and AP-1 by acetaldehyde is independent of oxidative stress.


HSP: Bystander Antigen in Atopic Diseases?
Over the last years insight in the complex interactions between innate and adaptive immunity in the regulation of an inflammatory response has increased enormously. This has revived the interest in stress proteins; proteins that are expressed during cell stress. As these proteins can attract and trigger an immunological response they can act as important mediators in this interaction. In this respect, of special interest are proteins that may act as modulators of both innate and adaptive immunity. Heat shock proteins (HSPs) are stress proteins that have these, and more, characteristics. More than two decades of studies on HSPs has revealed that they are part of intrinsic, "natural" mechanisms that steer inflammation. This has provoked comprehensive explorations of the role of HSPs in various human inflammatory diseases. Most studies have focused on classical autoimmune diseases. This has led to the development of clinical studies with HSPs that have shown promise in Phase II/III clinical trials. Remarkably, only very little is yet known of the role of HSPs in atopic diseases. In allergic disease a number of studies have investigated the possibility that allergen-specific regulatory T cell (Treg) function is defective in individuals with allergic diseases. This raises the question whether methods can be identified to improve the Treg repertoire. Studies from other inflammatory diseases have suggested HSPs may have such a beneficial effect on the T cell repertoire. Based on the immune mechanisms of atopic diseases, in this review we will argue that, as in other human inflammatory conditions, understanding immunity to HSPs is likely also relevant for atopic diseases. Specifically, we will discuss why certain HSPs such as HSP60 connect the immune response to environmental antigens with regulation of the inflammatory response. Thus they provide a molecular link that may eventually even help to better understand the immune pathological basis of the hygiene hypothesis.


N-terminal acetylation of the influenza ribonuclease PA-X promotes nuclear localization and host shutoff activity in a multifaceted manner.
To counteract host antiviral responses, influenza A virus triggers a global reduction of cellular gene expression, a process termed "host shutoff." A key effector of influenza A virus host shutoff is the viral endoribonuclease PA-X, which degrades host mRNAs. While many of the molecular determinants of PA-X activity remain unknown, a previous study identified a requirement for N-terminal acetylation of PA-X for its host shutoff activity, but did not address how this modification promotes activity. Here, we report that PA-X N-terminal acetylation has two functions that can be separated based on the position of the acetylation, on the first amino acid, i.e. the initiator methionine, vs. the second amino acid following initiator methionine excision. Modification at either site is sufficient to ensure correct PA-X localization to the nucleus. However, modification at the second amino acid is not sufficient for host shutoff activity of ectopically expressed PA-X, which requires N-terminal acetylation of the initiator methionine specifically. Interestingly, during infection, N-terminal acetylation of PA-X at any position results in PA-X host shutoff activity, and this is, in part, due to a functional interaction with the influenza protein NS1. This reveals an unexpected role for other viral proteins in PA-X activity. Our studies highlight the multifaceted nature of PA-X N-terminal acetylation in its ability to regulate the host shutoff activity of PA-X through multiple avenues.


Factors affecting marginal integrity of class II bulk-fill composite resin restorations.
Background. Bulk-fill composite resins are a new type of resin-based composite resins, claimed to have the capacity to be placed in thick layers, up to 4 mm. This study was carried out to evaluate factors affecting gap formation in Cl II cavities restored using the bulk-fill technique. Methods. A total of 60 third molars were used in this study. Two Cl II cavities were prepared in each tooth, one on the mesial aspect 1 mm coronal to the CEJ and one on the distal aspect 1 mm apical to the CEJ. The teeth were divided into 4 groups: A: The cavities were restored using the bulk-fill technique with Filtek P90 composite resin and its adhesive system and light-cured with quartz tungsten halogen (QTH) light-curing unit. B: The cavities were restored similar to that in group A but light-cured with an LED light-curing unit. C: The cavities were restored using the bulk-fill technique with X-tra Fil composite resin and Clearfil SE Bond adhesive system and light-cured with a QTH curing unit. D: The cavities were restored similar to that in group C but light-cured with an LED light-curing unit. The gaps were examined under a stereomicroscope at ×60. Data were analyzed with General Linear Model test. In cases of statistical significance (P<0.05), post hoc Bonferroni test was used for further analyses. Results. The light-curing unit type had no effect on gap formation. However, the results were significant in relation to the composite resin type and margin location (P<0.001). The cumulative effects of light-curing unit*gingival margin and light-curing unit*composite resin type were not significant; however, the cumulative effect of composite rein type*gingival margin was significant (P=0.04) Conclusion. X-tra Fil composite exhibited smaller gaps compared with Filtek P90 composite with both light-curing units. Both composite resins exhibited smaller gaps at enamel margins.


Mechanistic prediction of food effects for Compound A tablet using PBPK model.
Physiologically based pharmacokinetic (PBPK) modeling has been extensively used to study the factors of effect drug absorption, distribution, metabolize and extraction progress in human. In this study, Compound A(CPD A) is a BCS Class II drug, which has been extensive applied in clinical as lipid-lowering drug, administered orally after food, they displayed positive food effects in human, A PBPK model was built to mechanistic investigate the food effect of CPD A tablet in our study. By using gastroplus™ software, the PBPK models accurately predicted the results of food effects and predicted data were within 2-fold error of the observed results. The PBPK model mechanistic illuminated the changes of pharmacokinetic values for the positive food effects of the compound in human. Here in, the PBPK modeling which were combined with ACAT absorption models in it, successfully simulated the food effect in human of the drug. The simulation results were proved that PBPK model can be able to serve as a potential tool to predict the food effect on certain oral drugs.


Synthesis and structure-activity studies of a series of spirooxazolidine-2,4-diones: 4-oxa analogues of the muscarinic agonist 2-ethyl-8-methyl-2,8-diazaspiro[4.5]decane-1,3-dione.
A series of spirooxazolidine-2,4-dione derivatives related to the putative M1 agonist 2-ethyl-8-methyl-2,8-diazaspiro[4.5]decane-1,3-dione (RS86; 1) were synthesized. The compounds were evaluated as cholinergic agents in in vitro binding assays and in in vivo pharmacological tests including antiamnesic effects using scopolamine-treated mice, hypothermia, and salivation in mice. Four compounds (5a,c,f and 17a) exhibited affinity for cortical M1 receptors and reversed scopolamine-induced impairment of mouse passive avoidance tasks, as did 1. Among these compounds, only 5a exhibited M1-receptor stimulating activity in pithed rats. Structural requirements for muscarinic activity in this series of spirooxazolidine-2,4-dione derivatives were as strict as those reported for spirosuccinimide derivatives including 1. The antiamnesic dose of 3-ethyl-8-methyl-1-oxa-3,8-diazaspiro[4.5]decane-2,4-dione (5a) was 2 orders of magnitude lower than the doses inducing hypothermia and salivation, in contrast to 1 for which the former dose was only 5-10-fold lower than the latter. These results suggest that the 8-azaspiro[4.5]decane skeleton represents a useful template for designing new muscarinic agonists as antidementia drugs.


Radial Icicle Tree (RIT): Node Separation and Area Constancy.
Icicles and sunbursts are two commonly-used visual representations of trees. While icicle trees can map data values faithfully to rectangles of different sizes, often some rectangles are too narrow to be noticed easily. When an icicle tree is transformed into a sunburst tree, the width of each rectangle becomes the length of an annular sector that is usually longer than the original width. While sunburst trees alleviate the problem of narrow rectangles in icicle trees, it no longer maintains the consistency of size encoding. At different tree depths, nodes of the same data values are displayed in annular sections of different sizes in a sunburst tree, though they are represented by rectangles of the same size in an icicle tree. Furthermore, two nodes from different subtrees could sometimes appear as a single node in both icicle trees and sunburst trees. In this paper, we propose a new visual representation, referred to as radial icicle tree (RIT), which transforms the rectangular bounding box of an icicle tree into a circle, circular sector, or annular sector while introducing gaps between nodes and maintaining area constancy for nodes of the same size. We applied the new visual design to several datasets. Both the analytical design process and user-centered evaluation have confirmed that this new design has improved the design of icicles and sunburst trees without introducing any relative demerit.


Myelin transcription factor 1 (Myt1) expression in demyelinated lesions of rodent and human CNS.
Myelin transcription factor 1 (Myt1) is a zinc-finger DNA binding protein that influences developing oligodendrocyte progenitor (OP) cell proliferation, differentiation, and myelin gene transcription in vitro. The potential of Myt1 to play a role in OP responses leading to remyelination was examined using murine hepatitis virus strain A59 (MHV) to induce spinal cord demyelination and potential relevance to human pathology was evaluated in multiple sclerosis (MS) lesions. In MHV-infected mice, the density of Myt1 expressing cells markedly increased in lesioned areas of spinal cord white matter. Myt1 expressing cells proliferated most extensively during active demyelination and subsequently accumulated to maximal levels during early remyelination. Cells with nuclear Myt1 immunoreactivity were mainly OP cells, identified by co-localization with platelet-derived growth factor alpha receptor, with additional phenotypes being either oligodendrocytes or neural stem cells, identified by CC1 antigen and Musashi1, respectively. The density of OP cells expressing Myt1 was significantly increased in white matter of MHV-infected mice during demyelination and early remyelination then as remyelination advanced the values returned to levels comparable to PBS-injected control mice. In MHV lesions, Myt1 was not expressed in astrocytes, lymphocytes, or macrophage/microglial cells. MS lesions demonstrated increased Myt1 expression in both the periplaque white matter adjacent to lesions and within early remyelinating lesions. These results suggesta potential role for Myt1 in the regeneration of oligodendrocyte lineage cells in response to demyelination.


Accessory proteins for melanocortin signaling: attractin and mahogunin.
Switching from eumelanin to pheomelanin synthesis during hair growth is accomplished by transient synthesis of Agouti protein, an inverse agonist for the melanocortin-1 receptor (Mc1r). The coat color mutations mahogany and mahoganoid prevent hair follicle melanocytes from responding to Agouti protein. The gene mutated in mahogany, which is also known as Attractin (Atrn), encodes a type I transmembrane protein that functions as an accessory receptor for Agouti protein. We have recently determined that the gene mutated in mahoganoid, which is also known as Mahogunin (Mgrn1), encodes an E3 ubiquitin ligase. Like Attractin, Mahogunin is conserved in invertebrate genomes, and its absence causes a pleiotropic phenotype that includes spongiform neurodegeneration.


Deletion of zmp1 improves Mycobacterium bovis BCG-mediated protection in a guinea pig model of tuberculosis.
Having demonstrated previously that deletion of zinc metalloprotease zmp1 in Mycobacterium bovis BCG increased immunogenicity of BCG vaccines, we here investigated the protective efficacy of BCG zmp1 deletion mutants in a guinea pig model of tuberculosis infection. zmp1 deletion mutants of BCG provided enhanced protection by reducing the bacterial load of tubercle bacilli in the lungs of infected guinea pigs. The increased efficacy of BCG due to zmp1 deletion was demonstrated in both BCG Pasteur and BCG Denmark indicating that the improved protection by zmp1 deletion is independent from the BCG sub-strain. In addition, unmarked BCG Δzmp1 mutant strains showed a better safety profile in a CB-17 SCID mouse survival model than the parental BCG strains. Together, these results support the further development of BCG Δzmp1 for use in clinical trials.


Analysis of the lowest instrumented vertebra following anterior spinal fusion of thoracolumbar/lumbar adolescent idiopathic scoliosis: can we predict postoperative disc wedging?
A retrospective radiographic study. To investigate which radiographic parameters correlate best to ultimate lowest instrumented vertebra (LIV) position and subjacent disc wedging following anterior spinal fusion (ASF) for thoracolumbar/lumbar (TL/L) adolescent idiopathic scoliosis (AIS). In an ASF of TL/L AIS, part of the operative goals are often to horizontalize and centralize the LIV, or potentially minimize subjacent disc wedging after surgery. To our knowledge, no study has investigated the specific radiographic parameters involved with obtaining these goals. Sixty-one patients with TL/L AIS were treated with an instrumented ASF with a minimum 2-year follow-up. Preoperative and postoperative radiographs were examined measuring various radiographic parameters of the curve itself along with the LIV and subjacent disc. Specific correlation of these parameters to the coronal disc angle immediately below the LIV (disc angle), LIV translation, and global coronal balance (C7-CSVL distance) at 2 years postoperative was analyzed, respectively. The preoperative disc angle was 4.49 degrees +/- 5.48 and postoperative -5.85 degrees +/- 4.37. The change of the disc angle was significantly correlated to the LIV level relative to the preoperative lower end vertebra (LEV) (P < 0.006). Regressive analysis demonstrated the correlative parameters to the postoperative disc angle to be: preoperative upright disc angle; preoperative apex-LIV distance; and preoperative T12-LIV lordosis (P < 0.0001, r2 = 0.51). The correlative parameters to postoperative LIV translation were preoperative LIV translation and preoperative LIV rotation (P = 0.002, r2 = 0.2). The correlative parameter to postoperative C7-CSVL distance was only preoperative C7-CSVL distance (P < 0.0001, r2 = 0.3). Postoperative subjacent disc wedging occurs most often when the preoperative subjacent disc is nearly parallel and when a shorter fusion excluding the LEV is performed. Preoperative LIV rotation significantly correlates to postoperative LIV translation. Surgeons should note these preoperative predictive factors to optimize radiographic results of the operative treatment of TL/L AIS.


The value of sonographic microvascular imaging in the diagnosis of lipedema.
Lipedema is a chronic disease marked by symmetric enlargement of painful nodular and fibrotic adipose tissue, predominantly affecting the limbs. Since there is no specific test or biomarker for this condition, years often pass before the diagnosis of lipedema is established for the first time, thereby causing psychosocial distress, including depression, eating disorders, and social isolation. Over the last few years several advanced Doppler-based technologies have been developed to visualize slow flow blood vessels and superficial microvascular architecture undetectable by traditional color Doppler flow imaging (CDFI). The aim of this study was to evaluate the superficial microvascular anatomy in lipedema patients compared to healthy controls and investigate the clinical significance of the Ultra Micro Angiography (UMA) technology in the diagnosis of lipedema. This new technique may contribute to reduce the diagnostic delay and, eventually, establish and guide treatment strategies toward a better therapeutic outcome in lipedema patients. 25 patients with lipedema and ten healthy controls with no history of lipedema were included in this study. All ultrasound examinations were performed on a novel high-performance ultrasound system (Resona R9/Mindray) using CDFI and the UMA technique. In all of the patients, Ultra Micro Angiography achieved the excellent visualization of microvascular structures, revealing that most lipedema patients showed grade 3 (n = 13) or grade 2 (n = 8) flow. UMA was superior to CDFI for depicting the microvascular structures. Here we show that UMA imaging characterizes the subcutaneous microvasculature with an unprecedented accuracy. The method has the advantage of being sensitive to small, slow-flowing vessels. This allows for the assessment of the course of vessels and vascular pathologies in great detail. Thus, UMA as a non-invasive diagnostic method can improve diagnostic accuracy in lipedema.


Maternal mortality in South Africa in 2001: From demographic census to epidemiological investigation.
Maternal mortality remains poorly researched in Africa, and is likely to worsen dramatically as a consequence of HIV/AIDS. The 2001 census of South Africa included a question on deaths in the previous 12 months, and two questions on external causes and maternal mortality, defined as "pregnancy-related deaths". A microdata sample from the census permits researchers to assess levels and differentials in maternal mortality, in a country severely affected by high death rates from HIV/AIDS and from external causes. After correcting for several minor biases, our estimate of the Maternal Mortality Ratio (MMR) in 2001 was 542 per 100,000 live births. This level is much higher than previous estimates dating from pre-HIV/AIDS times. This high level occurred despite a relatively low proportion of maternal deaths (6.4%) among deaths of women aged 15-49 years, and was due to the astonishingly high level of adult mortality, some 4.7 times higher than expected from mortality below age 15 or above age 50. The main reasons for these excessive levels were HIV/AIDS and external causes of deaths. Our regional estimates of MMR were found to be consistent with other findings in the Cape Town area, and with the Agincourt DSS. The differentials in MMR were considerable: 1 to 9.2 for population groups (race), 1 to 3.2 for provinces, and 1 to 2.4 for levels of education. Relationship with income and wealth were complex, with highest values for middle income and middle wealth index. The effect of urbanization was small, and reversed in a multivariate analysis. Higher risks in provinces were not necessarily associated with lower income, lower education or higher proportions of home delivery, but correlated primarily with the prevalence of HIV/AIDS. Demographic census microdata offer the opportunity to conduct an epidemiologic analysis of maternal mortality. In the case of South Africa, the level of MMR increased dramatically over the past 10 years, most likely because of HIV/AIDS. Indirect causes of maternal deaths appear much more important than direct obstetric causes. The MMR appears no longer to be a reliable measure of the quality of obstetric care or a measure of safe motherhood.


Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation.
Neutrophils are the most abundant type of white blood cells. Upon stimulation, they are able to decondense and release their chromatin as neutrophil extracellular traps (NETs). This process (NETosis) is part of immune defense mechanisms but also plays an important role in many chronic and inflammatory diseases such as atherosclerosis, rheumatoid arthritis, diabetes, and cancer. For this reason, much effort has been invested into understanding biochemical signaling pathways in NETosis. However, the impact of the mechanical micro-environment and adhesion on NETosis is not well-understood. Here, we studied how adhesion and especially substrate elasticity affect NETosis. We employed polyacrylamide (PAA) gels with distinctly defined elasticities (Young's modulus E) within the physiologically relevant range from 1 to 128 kPa and coated the gels with integrin ligands (collagen I, fibrinogen). Neutrophils were cultured on these substrates and stimulated with potent inducers of NETosis: phorbol 12-myristate 13-acetate (PMA) and lipopolysaccharide (LPS). Interestingly, PMA-induced NETosis was neither affected by substrate elasticity nor by different integrin ligands. In contrast, for LPS stimulation, NETosis rates increased with increasing substrate elasticity (E > 20 kPa). LPS-induced NETosis increased with increasing cell contact area, while PMA-induced NETosis did not require adhesion at all. Furthermore, inhibition of phosphatidylinositide 3 kinase (PI3K), which is involved in adhesion signaling, completely abolished LPS-induced NETosis but only slightly decreased PMA-induced NETosis. In summary, we show that LPS-induced NETosis depends on adhesion and substrate elasticity while PMA-induced NETosis is completely independent of adhesion.


Three-dimensional excitation emission matrix fluorescence spectroscopy and gel-permeating chromatography to characterize extracellular polymeric substances in aerobic granulation.
Three-dimensional excitation emission matrix (EEM) fluorescence spectroscopy and gel-permeating chromatography (GPC) were employed to characterize the extracellular polymeric substances (EPS) in aerobic granulation. EPS matrix in this study was stratified into four fractions: (1) supernatant, (2) slime, (3) loosely bound EPS (LB-EPS), and (4) tightly bound EPS (TB-EPS). The results showed that the dissolved organic carbon was mainly distributed in TB-EPS fraction, and increased with increasing the operating time. The supernatant, slime, and LB-EPS fractions exhibited four fluorescence peaks, an autochthonous signature, unimodal MW distribution and lower molecular weight (MW) (3 < log [MW]<5), whereas the TB-EPS fraction only had two peaks, an allochthonous signature, multiple peaks and higher MW (5 < log [MW]<7). It was deemed that the formation of aerobic granules was correlated with the accumulation of proteins in the TB-EPS fraction. EEM spectroscopy and GPC profiles could be used as appropriate and effective methods to characterize the EPS in aerobic granulation from a micro-view level.


Trimethylamine-N-oxide (TMAO) as Novel Potential Biomarker of Early Predictors of Metabolic Syndrome.
There is a mechanistic link between the gut-derived metabolite trimethylamine-N-oxide (TMAO) and obesity-related diseases, suggesting that the TMAO pathway may also be linked to the pathogenesis of obesity. The Visceral Adiposity Index (VAI), a gender-specific indicator of adipose dysfunction, and the Fatty Liver Index (FLI), a predictor of non-alcoholic fatty liver disease (NAFLD), are early predictors of metabolic syndrome (MetS). In this cross-sectional observational study, we investigated TMAO levels in adults stratified according to Body Mass Index (BMI) and the association of TMAO with VAI and FLI. One hundred and thirty-seven adult subjects (59 males; 21⁻56 years) were enrolled. TMAO levels were detected using HPLC/MS analysis. Homeostatic Model Assessment of Insulin Resistance (HoMA-IR), VAI and FLI were included as cardio-metabolic indices. TMAO levels increased along with BMI and were positively associated with VAI and FLI, independently, on common potential covariates. The most sensitive and specific cut-offs for circulating levels of TMAO to predict the presence of NAFLD-FLI and MetS were ≥8.02 µM and ≥8.74 µM, respectively. These findings allow us to hypothesize a role of TMAO as an early biomarker of adipose dysfunction and NAFLD-FLI in all borderline conditions in which overt MetS is not present, and suggest that a specific cut-off of TMAO might help in identifying subjects at high risk of NAFLD.


Rapidly enlarging neck masses of the thyroid with Horner's syndrome: a concise clinical review.
Horner's syndrome (HS) presenting with a neck mass is a rare but challenging clinical scenario which may be caused by malignant thyroid disease. A concise review of the literature (PubMED database; 1990-2013) on the clinical management of neck masses with HS. An example case is also discussed. 1.3% of HS is caused by thyroid pathology. Thyroid pathology is the commonest cause of a neck mass associated with HS: the majority are caused by benign pathology; with carcinoma and lymphoma accounting for the remainder. Anaplastic thyroid carcinoma (ATC), thyroid lymphoma (TL) and thyroid sarcoma (TS) typically present with rapidly enlarging anterior neck masses in the elderly and are difficult to distinguish clinically. Although fine needle aspirate cytology (FNAC) is the diagnostic tool of choice for thyroid masses, core or incisional biopsy may be necessary when FNAC is inconclusive. Differentiation between ATC, TL and TS is imperative as their treatment and prognoses differ greatly. Where feasible a combination of surgical debulking, radiotherapy and chemotherapy is the treatment of choice in ATC. Advanced cases benefit from 2 monthly endoscopic surveillance ± tracheostomy, stenting or Nd-YAG laser therapy. Aggressive oncological resection alone is recommended in TS. Treatment regimes in thyroid lymphoma (typically chemotherapy ± radiotherapy) differ for histological sub-types. 5-year failure-free survival is 90% in TL compared with a mean survival of 6-8 months in ATC and 10 months in TS.


Palladium-catalyzed amination of aromatic C-H bonds with oxime esters.
We report a conceptually new approach to the direct amination of aromatic C-H bonds. In this process, an oxime ester function reacts with an aromatic C-H bond under redox-neutral conditions to form, in the case studied, an indole product. These reactions occur with relatively low catalyst loading (1 mol %) by a mechanism that appears to involve an unusual initial oxidative addition of an N-O bond to a Pd(0) species. The Pd(II) complex from oxidative addition of the N-X bond has been isolated for the first time, and evidence for the intermediacy of such oxidative addition products in the catalytic reaction has been gained.


Negative energy balance and leptin regulate neuromedin-U expression in the rat pars tuberalis.
Central neuromedin U (NMU) functions in energy balance, the hypothalamic-pituitary-adrenal axis, LH release and circadian rhythmicity. In rats, high levels of NMU occur in the hypothalamic suprachiasmatic nuclei and the pars tuberalis of the pituitary. NMU expression in the pars tuberalis appears to be downregulated in the Zucker fatty (fa/fa) rat, lacking functional leptin receptors. In contrast, in the dorsomedial (DMH) nuclei of the mouse, NMU expression is higher in the ob/ob mouse, lacking leptin, and is upregulated by fasting. However, leptin appears not to change NMU gene expression in either the mouse DMH or the rat pars tuberalis. Thus, the present study aims to better identify factors influencing central NMU expression in the rat pars tuberalis. Sprague-Dawley rats were fasted and/or challenged with intracerebroventricular leptin or ghrelin and gene expression was measured using real-time reverse transcriptase-PCR and quantitative in situ hybridisation with riboprobes specific for NMU and NMU receptor (NMU-R2). NMU expression in the rat pars tuberalis was elevated by fasting. Ghrelin administration had no effect on the level of NMU expression, but leptin was found to diminish the expression in a concentration- and time-dependent manner. NMU-R2 expression was unchanged in any of the groups measured. These results suggest that NMU expression in rat pars tuberalis is upregulated in states of negative energy balance, and this may be mediated indirectly by changes in leptin levels. These results demonstrate a link between energy balance and NMU expression in the pars tuberalis of the pituitary.


Cloning and characterization of mouse CCAAT binding factor.
Isolation of cDNA clones for the mouse CCAAT binding factor (mCBF) has revealed the expression of two distinct forms of mCBF that are generated by alternative splicing of a single primary transcript from a gene that maps to chromosome 17. The mCBF1 mRNA encodes a protein of 997 amino acids, whereas the mCBF2 protein is predicted to be only 461 amino acids in length; mCBF1 and human CBF (hCBF) share>80% amino acid sequence identity. Analysis of adult mouse tissue RNAs has revealed that the mCBF1 and mCBF2 mRNAs are ubiquitously expressed, but that mCBF1 mRNA is 5- to 10-fold more abundant than mCBF2 mRNA. Similarly, mCBF mRNA was detected through-out the placenta and in all tissues of the developing embryo from day 8 to day 18 of gestation. Overexpression of the two forms of mCBF in mammalian cells has demonstrated that the mCBF1 and mCBF2 proteins localize to different cellular compartments, with mCBF1 found predominantly in the nucleus and mCBF2 restricted to the cytoplasm. Co-expression of these two forms influences their localization, however, indicating that CBF activity can be regulated by the relative amounts of the two forms expressed in a cell.


Fast physical random bit generation with photonic integrated circuits with different external cavity lengths for chaos generation.
We generate random bit sequences from chaotic temporal waveforms by using photonic integrated circuits (PICs) with different external cavity lengths. We investigate the condition for generating random bits at different sampling rates of single-bit generation method with the PICs. We succeed in generating certified random bit sequences by using the PIC with 3, 4, 5, or 10-mm-long external cavity, whereas random bits cannot pass all the statistical tests of randomness when the PIC with 1 or 2 mm-long external cavity is used.


Shiga toxin-mediated disease in MyD88-deficient mice infected with Escherichia coli O157:H7.
Toll-like receptors (TLRs) are key factors of innate immunity that detect pathogen invasion and trigger a host response. TLR4 can mediate a response through adaptor molecules, MyD88 or TRIF. In the present study, streptomycin-treated MyD88(-/-), Tlr4(-/-), Trif (Lps2/Lps2), and C57BL/6 wild-type (WT) mice were infected with either Shiga toxin (Stx)-producing or non-producing Escherichia coli O157:H7. Moderate to severe clinical signs of disease developed in MyD88(-/-) (n = 21/21), Tlr4(-/-) (n = 12/16), Trif (Lps2/Lps2) (n = 7/15) and WT mice (n = 6/20) infected with Stx-producing E. coli O157:H7 but not in mice inoculated with the Stx non-producing strain (n = 0/54, P < 0.001). MyD88(-/-) mice infected with Stx-producing E. coli O157:H7 developed the most severe disease and had the highest bacterial burden. Hematological analysis of sick MyD88(-/-) mice showed reduced red blood cell counts and reticulocytosis, suggesting hemolysis. Thrombocytopenia developed in MyD88(-/-), Trif (Lps2/Lps2), and WT mice, and creatinine levels were elevated in both MyD88(-/-) and WT mice infected with the Stx-producing strain. Renal histopathology showed evidence of glomerular capillary congestion, tubular desquamation, and fibrinogen deposition, and intestinal histopathology showed mucosal injury, edema, and inflammation in sick mice. Administration of purified Stx2 to MyD88(-/-) and WT mice led to severe disease in both groups, suggesting that MyD88(-/-) mice are not more sensitive to Stx than WT mice. As MyD88(-/-) mice developed the most severe disease hematological and pathological changes, the results suggest that dysfunctional innate immune responses via MyD88 enhanced Stx-induced disease.


From one gene to two proteins: the biogenesis of cytochromes b and c1 in Bradyrhizobium japonicum.
Genes coding for polyproteins that are cleaved posttranslationally into two or more functional proteins are rarely found in prokaryotes. One example concerns the biogenesis of the Bradyrhizobium japonicum cytochromes b and c1, two of the three constituent subunits of ubiquinol-cytochrome-c reductase (ubiquinol:ferricytochrome-c oxidoreductase, EC 220.127.116.11); the respective apoproteins for these subunits are encoded by the 5' and 3' halves of a single gene, fbcH. These two halves are linked by an extra piece of DNA encoding a characteristic signal peptide for protein translocation across the cytoplasmic membrane. Processing of the fbcH gene product is shown to occur at a typical signal peptidase recognition site. This reaction is reminiscent of that catalyzed by the regular bacterial signal peptidase that normally cleaves off presequences from the N termini of translocated proteins. Mutational alteration of the signal peptidase recognition site within FbcH results in the appearance of an uncleaved bc1 fusion protein in the membrane. Additionally, a functional heme-binding site in the apocytochrome c1 section of FbcH is shown to be a necessary prerequisite for the formation of the bc1 complex.


Following intracellular cholesterol transport by linear and non-linear optical microscopy of intrinsically fluorescent sterols.
Elucidation of intracellular cholesterol transport is important for understanding the molecular basis of several metabolic and neuronal diseases, like atheroclerosis or lysosomal storage disorders. Progress in this field depends crucially on the development of new technical approaches to follow the cellular movement of this essential lipid molecule. In this article, a survey of the various methods being used for analysis of sterol trafficking is given. Various classical biochemical methods are presented and their suitability for analysis of sterol trafficking is assessed. Special emphasis is on recent developments in imaging technology to follow the intracellular fate of intrinsically fluorescent sterols as faithful cholesterol markers. In particular, UV-sensitive wide field and multiphoton microscopy of the sterol dehydroergosterol, DHE, is explained and new methods of quantitative image analysis like pixel-wise bleach rate fitting and multiphoton image correlation spectroscopy are introduced. Several applications of the new technology including observation of vectorial sterol trafficking in polarized human hepatoma cells for investigation of reverse cholesterol transport are presented.


[Effect of riodoxol on herpes simplex virus morphogenesis in experimental eye infection].
The effect of riodoxol on herpes simplex type 1 viruses was studied. It was shown to affect multiplication and maturation of the virus as manifested in the appearance of virions with incomplete nucleoid structure and long-term presence of virions in cytoplasmic vacuoles in cells. Submicroscopic changes suggest a dual effect of riodoxol on herpes simplex virus: on the DNA replicative cycle and on the stage of late maturation of virus at the time of its release from the nucleus.'


Modulation of hepatic acute phase gene expression by epidermal growth factor and Src protein tyrosine kinases in murine and human hepatic cells.
As part of systemic inflammatory reactions, interleukin 6 (IL-6) induces acute phase protein (APP) genes through the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. Epidermal growth factor (EGF), which contributes to the regenerative process after liver injury and also activates STATs, does not induce but attenuates IL-6-stimulated expression of several APP genes in primary mouse hepatocytes. The APP-modifying action of EGF receptor (EGFR) was characterized in HepG2 cells. Although EGF less effectively engages STAT proteins in these cells, it reduces expression of fibrinogen and haptoglobin, but stimulates production of alpha(1)-antichymotrypsin and induces transcription through the alpha(1)-antichymotrypsin and C-reactive protein promoter. The stimulatory EGFR signal is insensitive to inhibition of JAKs and appears to involve Src kinases and STAT proteins as shown by inhibition through overexpression of C-terminal Src kinase (Csk) and transdominant negative STAT3, respectively. A mediator role of Src is supported by the ability of c-Src and v-Src to activate STATs and induce transcription through APP promoters. Src kinases have been observed in association with the IL-6 receptor; however, inhibition of Src kinases by Csk enhances IL-6-induced transcription. The Csk effect is attributed to prevention of Src kinases from phosphorylating gp130 at the docking site for the signal-moderating protein tyrosine phosphatase SHP-2. The inhibitory EGFR signal on APP expression correlates with the activation of Erk1 and Erk2. The study shows a dual signaling function for EGFR and suggests that the ratio of receptor-activated STATs and Erks influence the level of stimulated or inhibited expression of individual APPs.


A system for encoding and searching Markush structures.
The encoding and searching of generic chemical structures, so-called Markush structures, have received little attention in the literature of late. The ability to encode and search these complex entities is of use in various branches of chemoinformatics. We describe a general language for encoding Markush structures and algorithms for searching them and give three examples of the utility of such a system: development of general Free-Wilson analyses of chemical series, detection of controlled substances within a large database of molecular structures, and searching of large databases of virtual compounds.


Light-induced changes in the chemical bond structure of light-harvesting complex II probed by FTIR spectroscopy.
Light-harvesting complex II (LHC-II) regulates the light energy distribution between photosystem I and II in plants. This process is mediated by phosphorylation of the LHC-II protein, which depends on the oxidation state of photosynthetic electron carriers. In addition to this regulatory mechanism, it has recently been proposed that light can directly induce a conformational change in isolated LHC-II. To provide biophysical evidence for such a conformational change in the protein, we studied infrared absorbance changes in isolated LHC-II upon exposure to light flashes. Compared to the signals obtained with other proteins that exhibit well-characterized conformational changes, the signal in the LHC-II difference spectra is very weak. The position of the difference bands coincides with the main IR absorption bands of chlorophyll. We conclude that there are no detectable light-induced changes in the LHC protein structure and attribute the observed IR signals to light-induced chlorophyll degradation.


Prognostic value of asymptomatic skin sensitization to aeroallergens.
Asymptomatic skin sensitization to aeroallergens is frequently encountered in epidemiological studies and in everyday clinical life. Correct management of the condition is essential to avoid both progression into allergic disease and unnecessary intervention. Understanding immunological mechanisms in asymptomatic skin sensitization might provide new insights into the natural history and treatment of respiratory allergy. Research on asymptomatic skin sensitization is rare, and the present review unites previous studies with recent findings. It is a common condition affecting 8-30% of the population when using a local standard panel of aeroallergens. Clinically, immediate but not late-phase reactions are induced by allergen challenge. Absent eosinophil stimulation and migration and low IL-5 levels appear to be sentinel mechanisms. Prospective studies show that 30-60% become allergic, depending on allergens and follow-up period. No prospective intervention studies have been performed; however, allergen avoidance seems efficacious in reducing allergy development to occupational and domestic allergens. Asymptomatic skin sensitization due to an erroneously positive skin test must be ruled out before allergen avoidance measures are initiated. Surprisingly few papers exist on asymptomatic skin sensitization epidemiology and immunology, despite the intriguing question as to why symptoms do not develop in IgE-sensitized patients. It is a common condition and a risk factor for later development of respiratory allergic disease. Cross-sectional intervention studies suggest that allergy development is reduced by allergen avoidance. Immunologically, control of eosinophil stimulation and migration seems to be pivotal. How this control is maintained remains to be elucidated.


Bio-inspired incorporation of phenylalanine enhances ionic selectivity in layer-by-layer deposited polyelectrolyte films.
The addition of a common amino acid, phenylalanine, to a Layer-by-Layer (LbL) deposited polyelectrolyte (PE) film on a nanoporous membrane can increase its ionic selectivity over a PE film without the added amino acid. The addition of phenylalanine is inspired by detailed knowledge of the structure of the channelrhodopsins family of protein ion channels, where phenylalanine plays an instrumental role in facilitating sodium ion transport. The normally deposited and crosslinked PE films increase the cationic selectivity of a support membrane in a controllable manner where higher selectivity is achieved with thicker PE coatings, which in turn also increases the ionic resistance of the membrane. The increased ionic selectivity is desired while the increased resistance is not. We show that through incorporation of phenylalanine during the LbL deposition process, in solutions of NaCl with concentrations ranging from 0.1 to 100 mM, the ionic selectivity can be increased independently of the membrane resistance. Specifically, the addition is shown to increase the cationic transference of the PE films from 81.4% to 86.4%, an increase on par with PE films that are nearly triple the thickness while exhibiting much lower resistance compared to the thicker coatings, where the phenylalanine incorporated PE films display an area specific resistance of 1.81 Ω cm2 in 100 mM NaCl while much thicker PE membranes show a higher resistance of 2.75 Ω cm2 in the same 100 mM NaCl solution.


Controlled activation of morphogenesis to generate a functional human microvasculature in a synthetic matrix.
Understanding the role of the extracellular matrix (ECM) in vascular morphogenesis has been possible using natural ECMs as in vitro models to study the underlying molecular mechanisms. However, little is known about vascular morphogenesis in synthetic matrices where properties can be tuned toward both the basic understanding of tubulogenesis in modular environments and as a clinically relevant alternative to natural materials for regenerative medicine. We investigated synthetic, tunable hyaluronic acid (HA) hydrogels and determined both the adhesion and degradation parameters that enable human endothelial colony-forming cells (ECFCs) to form efficient vascular networks. Entrapped ECFCs underwent tubulogenesis dependent on the cellular interactions with the HA hydrogel during each stage of vascular morphogenesis. Vacuole and lumen formed through integrins α(5)β(1) and α(V)β(3), while branching and sprouting were enabled by HA hydrogel degradation. Vascular networks formed within HA hydrogels containing ECFCs anastomosed with the host's circulation and supported blood flow in the hydrogel after transplantation. Collectively, we show that the signaling pathways of vascular morphogenesis of ECFCs can be precisely regulated in a synthetic matrix, resulting in a functional microvasculature useful for the study of 3-dimensional vascular biology and toward a range of vascular disorders and approaches in tissue regeneration.


[Biochemical mechanisms of acquired radioresistance of tumor tissue].
The level of lipoperoxidation products in radioresistant tumour is 3 times lower and superoxide dismutase activity--3.5 times higher than in the intact tumour. That is an evidence of high stability. The level of lipoperoxidation products in the brain and liver of rats with radioresistant variant of Guerin carcinoma is essentially less and catalase activity--considerably higher, than in the rats with intact tumour. This evidences for the corresponding increase of antioxidant protection of the organisms in the process of acquiring the tumour radioresistance.


Role of the proteome in phytohormonal signaling.
Phytohormones are orchestrators of plant growth and development. A lot of time and effort has been invested in attempting to comprehend their complex signaling pathways but despite success in elucidating some key components, molecular mechanisms in the transduction pathways are far from being resolved. The last decade has seen a boom in the analysis of phytohormone-responsive proteins. Abscisic acid, auxin, brassinosteroids, cytokinin, ethylene, gibberellins, nitric oxide, oxylipins, strigolactones, salicylic acid--all have been analyzed to various degrees. For this review, we collected data from proteome-wide analyses resulting in a list of over 2000 annotated proteins from Arabidopsis proteomics and nearly 500 manually filtered protein families merged from all the data available from different species. We present the currently accepted model of phytohormone signaling, highlight the contributions made by proteomic-based research and describe the key nodes in phytohormone signaling networks, as revealed by proteome analysis. These include ubiquitination and proteasome mediated degradation, calcium ion signaling, redox homeostasis, and phosphoproteome dynamics. Finally, we discuss potential pitfalls and future perspectives in the field. This article is part of a Special Issue entitled: Plant Proteomics--a bridge between fundamental processes and crop production, edited by Dr. Hans-Peter Mock.


Pediatric Thoracic Trauma Mortality in Iraq and Afghanistan Compared to the United States National Trauma Data Bank.
The authors compared pediatric thoracic patients in the Joint Theatre Trauma Registry (JTTR) to those in the National Trauma Data Bank (NTDB) to assess differences in patient mortality rates and mortality risk accounting for age, injury patterns, and injury severity. Patients less than 19 years of age with thoracic trauma were identified in both the JTTR and NTDB. Multiple logistic regression, χ2, Student's t-test, or Mann-Whitney U test were used as indicated to compare the two groups. Pediatric thoracic trauma patients seen in Iraq and Afghanistan (n = 955) had a significantly higher mortality rate (15.1 vs. 6.0%, P <.01) than those in the NTDB (n = 9085). After controlling for covariates between the JTTR and the NTDB, there was no difference in mortality (odds ratio for mortality for U.S. patients was 0.74, 95% CI 0.52-1.06, P = .10). The patients seen in Iraq or Afghanistan were significantly younger (8 years old, interquartile ratio (IQR) 2-13 vs. 15, IQR 10-17, P <.01) had greater severity of injuries (injury severity score 17, IQR 12-26 vs. 12, IQR 8-22, P <.01), had significantly more head injuries (29 vs. 14%, P <.01), and over half were exposed to a blast. Pediatric patients with thoracic trauma in Iraq and Afghanistan in the JTTR had similar mortality rates compared to the civilian population in the NTDB after accounting for confounding covariates. These findings indicate that deployed military medical professionals are providing comparable quality of care in extremely challenging circumstances. This information has important implications for military preparedness, medical training, and casualty care.


Phase II trial of high-dose cisplatin plus etoposide plus vinblastine in non-small-cell lung cancer. A Hoosier Oncology Group study.
Fifty-one patients with advanced non-small-cell lung cancer were treated on a Hoosier Oncology Group protocol with an aggressive, high-dose cisplatin combination chemotherapy regimen. All patients had a Karnofsky performance status of 80% or higher and had no prior chemotherapy. The drug regimen consisted of cisplatin 30 mg/m2 days one through five, etoposide 40 mg/m2 days one through five, and vinblastine 5 mg/m2 day one. Therapy was given every three weeks for a total of three courses. Forty-five patients were evaluable for response and an objective response was seen in 15 patients (33%) with only one complete responder. The median duration of response was 16.5 weeks. The median survival for the entire group was 29.0 weeks. Toxicity was moderately severe with two treatment-related deaths (4%). Despite an aggressive chemotherapy regimen in a favorable patient population, there was no obvious evidence of a major therapeutic value.


Lon protease of the alpha-proteobacterium Agrobacterium tumefaciens is required for normal growth, cellular morphology and full virulence.
The ATP-dependent Lon (La) protease is ubiquitous in nature and regulates a diverse set of physiological responses in bacteria. In this paper a lon mutant of the alpha-proteobacterium Agrobacterium tumefaciens C58 has been characterized. Unlike lon mutants of Escherichia coli, the lon mutant of A. tumefaciens grows very slowly, is not filamentous and exhibits normal resistance to UV irradiation. The mutant retains motility and chemotaxis, produces apparently normal amounts of exopolysacchride, but displays severe defects in cell morphology, with 80 % of the mutant cells appearing Y-shaped. Lon protease of A. tumefaciens shares high homology with its counterparts in E. coli and in Sinorhizobium meliloti, and functionally complements an E. coli lon mutant for defects in morphology and RcsA-mediated regulation of capsular polysaccharide production. Mutations at sites of Lon(At) corresponding to the ATP-binding site and the active site serine of the E. coli Lon protease abolish complementation of phenotypes of the A. tumefaciens and E. coli lon mutants. The nucleotide sequence upstream of A. tumefaciens lon contains an element similar to the consensus sigma(32) heat-shock promoter of E. coli. Northern and Western blot analyses indicated that expression of lon is induced by elevated temperature, albeit to a much lower level than that of groEL. The lon mutant is highly attenuated for virulence, suggesting that Lon may be required for the proper expression, assembly or function of the VirB/D4-mediated T-DNA transfer system.


Genomic Studies of Local Adaptation in Natural Plant Populations.
Local adaptation arises as a result of selection by the local environment favoring phenotypes that enhance fitness. Geographic patterns of phenotypic variation are in part due to this selective process. Classically, the genetic basis of those phenotypes has been studied in plant populations using a quantitative genetic approach in which plants from different source populations are grown in common environments, in reciprocal transplant experiments, or in studies across a wide geographic and environmentally heterogeneous area. Limitations of these approaches to understanding the genetic basis of phenotypic variation can now be addressed with next generation sequencing, gene expression profiles, and epigenetic analysis. In this paper, I summarize contemporary genomic research on local adaptation by comparing findings from the Arabidopsis annual plant model system with long-lived tree species in four kinds of local adaptation studies: 1) genomic studies of transplant experiments; 2) landscape genomic studies; 3) gene expression studies; 4) epigenetic studies of local adaptation. Although the basic study designs of common garden, reciprocal transplants, and geographic variation have remained constant, the inclusion of contemporary genomic approaches has provided substantive advances in our understanding of the genetic underpinnings of local adaptation, including the impact of climate, the identification of candidate genes involved in genotype-by-environment interactions, and evidence for the potential role of epigenetic modification. Despite these advances, new questions are arising and key areas for future research include more exploration of gene networks in response to biotic and abiotic stressors and improved statistical tools for traits with polygenic inheritance.


Mutagenic action of isocyanates used in the production of polyurethanes.
Isocyanates used in the production of polyurethanes were investigated for mutagenic action in Salmonella typhimurium. These investigations showed that the most commonly used isocyanates, toluene diisocyanate (TDI) and 4,4'-methylene-diphenylisocyanate (MDI), are mutagenic. This effect can be ascribed to the amine analogues formed during the hydrolysis of isocyanates: TDI is mutagenic in TA 1538 and TA 98 after metabolic activation. This finding agrees with the results obtained with the amine analogue 2,4-toluenediamine. MDI, like the amine analogue 4,4'-methylenedianiline, is mutagenic in TA 100 after metabolic activation by rat liver enzymes (S-9 mix). A prepolymerized polyisocyanate of the MDI type is also mutagenic in assays using TA 100 and S-9 mix. It is concluded that isocyanates are potentially mutagenic and carcinogenic to man. In view of their widespread use in the work environment and in light of the high production figures for polyurethanes in industrialized countries, isocyanates must be considered to represent a serious health hazard.


Hypoxia-inducible factor 1a is a Tsc1-regulated survival factor in newborn neurons in tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is a genetic disorder caused by mutations in TSC1 or TSC2 resulting in hyperactivity of the mammalian target of rapamycin and disabling brain lesions. These lesions contain misplaced neurons enriched in hypoxia-inducible factor 1a (HIF1a). However, the relationship between TSC1/2 and HIF1a and the function of HIF1a in TSC neurons remain unexplored. Here, we examine the degree of HIF1a activity and its function in newborn Tsc1(null) neurons in a mouse model of TSC. Using single cell electroporation in the neurogenic subventricular zone (SVZ) of neonatal mice, we deleted Tsc1 and generated olfactory lesions containing misplaced Tsc1(null) neurons as previously reported. These newborn neurons displayed elevated HIF1a-mediated transcriptional activity when compared with Tsc1 heterozygote neurons and a marked resistance to cell death induced by a HIF1a antagonist. Electroporation of Hif1a targeting short hairpin RNA (shRNA) or dominant negative HIF1a constructs resulted in 80-90% loss of Tsc1(null) newborn neurons although sparing SVZ stem cells. Consistent with this later finding, induction of Hif1a shRNA expression during synaptic integration thus bypassing neuron production also resulted in newborn neuron death. Collectively, these results suggest that HIF1a acts as a molecular determinant of newborn neuron survival and that its TSC1-dependent up-regulation gave Tsc1(null) neurons a survival advantage, despite their misplacement in a novel microenvironment.


Compressed sensing for real-time energy-efficient ECG compression on wireless body sensor nodes.
Wireless body sensor networks (WBSN) hold the promise to be a key enabling information and communications technology for next-generation patient-centric telecardiology or mobile cardiology solutions. Through enabling continuous remote cardiac monitoring, they have the potential to achieve improved personalization and quality of care, increased ability of prevention and early diagnosis, and enhanced patient autonomy, mobility, and safety. However, state-of-the-art WBSN-enabled ECG monitors still fall short of the required functionality, miniaturization, and energy efficiency. Among others, energy efficiency can be improved through embedded ECG compression, in order to reduce airtime over energy-hungry wireless links. In this paper, we quantify the potential of the emerging compressed sensing (CS) signal acquisition/compression paradigm for low-complexity energy-efficient ECG compression on the state-of-the-art Shimmer WBSN mote. Interestingly, our results show that CS represents a competitive alternative to state-of-the-art digital wavelet transform (DWT)-based ECG compression solutions in the context of WBSN-based ECG monitoring systems. More specifically, while expectedly exhibiting inferior compression performance than its DWT-based counterpart for a given reconstructed signal quality, its substantially lower complexity and CPU execution time enables it to ultimately outperform DWT-based ECG compression in terms of overall energy efficiency. CS-based ECG compression is accordingly shown to achieve a 37.1% extension in node lifetime relative to its DWT-based counterpart for "good" reconstruction quality.


Postpartum Circulating Cell-Free Insulin DNA Levels Are Higher in Women with Previous Gestational Diabetes Mellitus Who Develop Type 2 Diabetes in Later Life.
Women with previous gestational diabetes mellitus (GDM) have evidence of postpartum β-cell dysfunction, which increases their risk of developing type 2 diabetes (T2DM) later in life. Elevated levels of circulating cell-free preproinsulin (INS) DNA correlate with dying β-cells in both mice and humans. The aim of this study was to determine if cell-free circulating INS DNA levels are higher in women with previous GDM who develop T2DM. We used droplet digital (dd) PCR to measure the levels of cell-free circulating methylated and unmethylated INS DNA in plasma from 97 women with normal glucose tolerance (NGT), 12 weeks following an index GDM pregnancy. Women were assessed for up to 10 years for the development of T2DM. In the follow-up period, 22% of women developed T2DM. Compared with NGT women, total cell-free INS DNA levels were significantly higher in women who developed T2DM (P = 0.02). There was no difference in cell-free circulating unmethylated and methylated INS DNA levels between NGT women and women who developed T2DM (P = 0.09 and P = 0.07, respectively). In women with a previous index GDM pregnancy, postpartum levels of cell-free circulating INS DNA are significantly higher in those women who later developed T2DM.


[Study on nurse performance evaluation: used methodology and obtained results - Part II].
In this second part of this study the methodology, the instruments utilized for the data gathering and the criteria used in the discussion of the date are presented. Part of the discussion of the results, which, for better comprehension, follow a pre-established path, is also presented. The results concerning the itens "objectives of the performance evaluation programs" and "elements responsible for the performance evaluation" are discussed. This last item is subdivided into "profissional characterization", which is totally discussed in this part of the study, and "specific professional skill" which will be presented in the third part of this paper, The theoretical background related to the results is contained in the discussion presented.


[Determination of groups at risk for acute intestinal infections by a population questionnaire].
Questionnaires were distributed in 3308 families divided into two groups: group 1 with no cases of acute intestinal infections and group 2 where such cases were registered. Information contained in the questionnaires was processes by means of computers bzsm-6. The analysis of the data on the occurrence of characteristics indicating the quality of water supply and water consumption (25 characteristics), sewage and sanitation (12 characteristics), living conditions (19 characteristics) showed that the living conditions of the families in group 2 were worse than those of the families in group 1. The occurrence of these characteristics in the families of patients with acute intestinal infections and in the families of carriers were mostly the same, and the existing differences in such characteristics as "fishing", "use of water from ponds for house-hold purposes" proved to be nonessential. The population of the city was divided into four risk groups with regard to the possibility of contacting infection, depending on the conditions of water supply, water consumption and sewage in different housing areas: the group registered as stable on account of sanitary and hygienic conditions, the groups of usual, increased and maximum risk.


Characterization of the mutS-proximal region of the Salmonella typhimurium SPI-1 identifies a group of pathogenicity island-associated genes.
The virulence properties of Salmonella enterica are largely encoded within a set of horizontally acquired gene blocks termed pathogenicity islands. One such pathogenicity island, SPI-1, located at centisome 63 of the Salmonella chromosome between the mutS and fhlA genes, encodes a type III protein secretion system and an iron uptake system. We have characterized the mutS-proximal border of this pathogenicity island and have identified two sets of genes, pigAB and pigCD. All four genes have homologs of unknown function in several bacteria that share the ability to establish an intimate association with higher eukaryotic hosts. The expression of at least two of these genes, pigA and pigB, is controlled by SprA, a transcription factor encoded within SPI-1 that controls the expression of genes associated with the type III secretion system of this island. In addition, we found that homologs of the pig genes are also found at different locations of the S. enterica chromosome in association with segments of DNA that exhibit features of pathogenicity islands. The presence of several apparently functional copies of these genes argues for an important role in the biology of this bacterial pathogen. Furthermore, they constitute a valuable tool to identify potential pathogenicity islands.


A clinical approach to common electrolyte problems: 3. Hypophosphatemia.
Hypophosphatemia is a common disorder caused by decreased intake, increased loss or transcellular shift of phosphorus. Symptoms of severe hypophosphatemia include reversible depression of myocardial function, acute respiratory failure, coma, rhabdomyolysis, osteomalacia, renal tubular acidosis and hemolysis. This paper discusses common clinical disorders associated with hypophosphatemia and presents an approach to diagnosis and treatment.


Influence of noradrenalin on pterygopalatine ganglionic transmission of the rabbit in vitro.
The influence of noradrenalin on pterygopalatine ganglionic transmission was examined in the rabbit in vitro. Electrical activity recorded from the posterior nasal nerve in response to stimulation of the vidian nerve was reversibly inhibited by the superfusion of noradrenalin. This inhibition was counteracted by phentolamine. This result indicates that, through alpha-adrenergic receptors, noradrenalin inhibits transmission in the pterygopalatine ganglion.


A Randomized Trial of Binocular Dig Rush Game Treatment for Amblyopia in Children Aged 4 to 6 Years.
Binocular treatment for unilateral amblyopia is an emerging treatment that requires evaluation through a randomized clinical trial. This study aimed to compare change in amblyopic-eye visual acuity (VA) in children aged 4 to 6 years treated with the dichoptic binocular iPad (Apple, Cupertino, CA) game, Dig Rush (not yet commercially available; Ubisoft, Montreal, Canada), plus continued spectacle correction versus continued spectacle correction alone. Children (mean age, 5.7 years) were randomly assigned to home treatment for 8 weeks with the iPad game (prescribed 1 h/d, 5 d/wk [n = 92], or continued spectacle correction alone [n = 90]) in a multicenter randomized clinical trial. Before enrollment, children wearing spectacles were required to have at least 16 weeks of wear or no improvement in amblyopic-eye VA (<0.1 logMAR) for at least 8 weeks. Outcome was change in amblyopic-eye VA from baseline to 4 weeks (primary) and 8 weeks (secondary) assessed by masked examiner. A total of 182 children with anisometropic (63%), strabismic (16%; <5∆ near, simultaneous prism and cover test), or combined-mechanism (20%) amblyopia (20/40 to 20/200; mean, 20/63) were enrolled. After 4 weeks, mean amblyopic VA improved by 1.1 logMAR lines with binocular treatment and 0.6 logMAR lines with spectacles alone (adjusted difference, 0.5 lines; 95.1% confidence interval [CI], 0.1 to 0.9). After 8 weeks, results (binocular treatment: mean amblyopic-eye VA improvement, 1.3 vs. 1.0 logMAR lines with spectacles alone; adjusted difference, 0.3 lines; 98.4% CI, -0.2 to 0.8 lines) were inconclusive because the CI included both zero and the pre-defined difference in mean VA change of 0.75 logMAR lines. In 4- to 6-year-old children with amblyopia, binocular Dig Rush treatment resulted in greater improvement in amblyopic-eye VA for 4 weeks but not 8 weeks. Future work is required to determine if modifications to the contrast increment algorithm or other aspects of the game or its implementation could enhance the treatment effect.


Two subtypes of enteric non-opioid sigma receptors in guinea-pig cholinergic motor neurons.
In the longitudinal muscle-myenteric plexus preparation (LMMP) of the guinea-pig ileum, the non-opioid sigma receptors agonists, 1,3-di-ortho-tolylguanidine (DTG) and (+)N-allyl-N-normetazocine [(+)SKF 10,047], had opposite effects on nerve-mediated cholinergic contractions caused by electrical field stimulation. DTG (0.1-10 microM) inhibited and (+)SKF 10,047 (0.1-10 microM) markedly enhanced these contractile responses. Both effects were evaluated in the presence (0.5 or 1 microM) of the putative antagonists at central sigma sites: haloperidol, rimcazole, BMY 14802 and dextromethorphan. Haloperidol and dextromethorphan were ineffective. Rimcazole antagonized the effect of both DTG and (+)SKF 10.047. BMY 14802 antagonized the (+)SKF 10.047-mediated excitatory response only. These results suggest that two sigma receptor subtypes are present in enteric cholinergic motor neurons innervating the longitudinal coat. Rimcazole and BMY 14802 may provide useful tools for the characterization of peripheral non-opioid sigma receptors.


Reversal of neurological deficits by painless nerve growth factor in a mouse model of Rett syndrome.
Rett syndrome is a rare genetic neurodevelopmental disease, affecting 1 in over 10,000 females born worldwide, caused by de novo mutations in the X-chromosome-located methyl-CpG-binding protein 2 (MeCP2) gene. Despite the great effort put forth by the scientific community, a therapy for this devastating disease is still needed. Here, we tested the therapeutic effects of a painless mutein of the Nerve Growth Factor, called human NGF painless (hNGFp), via a non-invasive intranasal delivery in female MeCP2+/- mice. Of note, previous work had demonstrated a broad biodistribution of hNGFp in the mouse brain by the nasal delivery route. We report that (1) the long-term lifelong treatment of MeCP2+/- mice with hNGFp, starting at 2 months of age, increased the chance of survival while also greatly improving behavioral parameters. Furthermore, when we assessed the phenotypic changes brought forth by (2) a short-term 1-month-long hNGFp-treatment, starting at 3 months of age (right after the initial presentation of symptoms), we observed the rescue of a well-known neuronal target population of NGF, cholinergic neurons in the medial septum. Moreover, we reveal a deficit in microglial morphology in MeCP2+/- mice, completely reversed in treated animals. This effect on microglia is in line with reports showing microglia to be a TrkA-dependent non-neuronal target cell population of NGF in the brain. To understand the immunomodulatory activity of hNGFp, we analyzed the cytokine profile after hNGFp treatment in MeCP2+/- mice, to discover that the treatment recovered the altered expression of key neuroimmune-communication molecules, such as fractalkine. The overall conclusion is that hNGFp delivered intranasally can ameliorate symptoms in the MeCP2+/- model of Rett syndrome, by exerting strong neuroprotection with a dual mechanism of action: directly on target neurons and indirectly via microglia.


Development and Validation of the CANHEART Population-Based Laboratory Prediction Models for Atherosclerotic Cardiovascular Disease.
Prediction of atherosclerotic cardiovascular disease (ASCVD) in primary prevention assessments exclusively with laboratory results may facilitate automated risk reporting and improve uptake of preventive therapies. To develop and validate sex-specific prediction models for ASCVD using age and routine laboratory tests and compare their performance with that of the pooled cohort equations (PCEs). Derivation and validation of the CANHEART (Cardiovascular Health in Ambulatory Care Research Team) Lab Models. Population-based cohort study in Ontario, Canada. A derivation and internal validation cohort of adults aged 40 to 75 years without cardiovascular disease from April 2009 to December 2015; an external validation cohort of primary care patients from January 2010 to December 2014. Age and laboratory predictors measured in the outpatient setting included serum total cholesterol, high-density lipoprotein cholesterol, triglycerides, hemoglobin, mean corpuscular volume, platelets, leukocytes, estimated glomerular filtration rate, and glucose. The ASCVD outcomes were defined as myocardial infarction, stroke, and death from ischemic heart or cerebrovascular disease within 5 years. Sex-specific models were developed and internally validated in 2 160 497 women and 1 833 147 men. They were well calibrated, with relative differences less than 1% between mean predicted and observed risk for both sexes. The c-statistic was 0.77 in women and 0.71 in men. External validation in 31 697 primary care patients showed a relative difference less than 14% and an absolute difference less than 0.3 percentage points in mean predicted and observed risks for both sexes. The c-statistics for the laboratory models were 0.72 for both sexes and were not statistically significantly different from those for the PCEs in women (change in c-statistic, -0.01 [95% CI, -0.03 to 0.01]) or men (change in c-statistic, -0.01 [CI, -0.04 to 0.02]). Medication use was not available at the population level. The CANHEART Lab Models predict ASCVD with similar accuracy to more complex models, such as the PCEs. Canadian Institutes of Health Research.


Morphologic evidence of photoreceptor differentiation of pinealocytes in the neonatal rat.
The pineal body and the retina of the neonatal Sprague-Dawley rat were studied by light and electron microscopy, and the morphologic differentiation of the parenchymal cells of the pineal body was compared with that of the developing photoreceptor cells of the retina. Between the ages of 4 and 12 days after birth, some of the developing pinealocytes were observed to become elongated and polarized, with their nuclei located at one pole. "Synaptic" ribbons were observed within the cell body. At the opposite pole the cells developed elongated cell processes that initially contained microtubules and ribosomes. These cell processes projected into luminal spaces and were attached by structures resembling zonulae adherentes to the adjacent cells. Extending from the tips of the cell processes, cilia with a 9 + 0 arrangement were observed. Lamellated and vesicular membranes were noted at the tips of the cilia. Such morphologic differentiation, however, could be observed only in rats younger than 17 days. Comparison of the morphologic features of the neonatal pinealocytes with those of the developing retinal photoreceptor cells showed much similarity. It is suggested that the pinealocytes of the neonatal rat undergo "photoreceptor-like" differentiation during a transient neonatal period. Such morphologic differentiation may provide an explanation for light-induced biochemical changes described in neonatal rats whose eyes had been enucleated.


Scoring System to Triage Patients for Spine Surgery in the Setting of Limited Resources: Application to the Coronavirus Disease 2019 (COVID-19) Pandemic and Beyond.
As of May 4, 2020, the coronavirus disease 2019 (COVID-19) pandemic has affected >3.5 million people and touched every inhabited continent. Accordingly, it has stressed health systems worldwide, leading to the cancellation of elective surgical cases and discussions regarding health care resource rationing. It is expected that rationing of surgical resources will continue even after the pandemic peak and may recur with future pandemics, creating a need for a means of triaging patients for emergent and elective spine surgery. Using a modified Delphi technique, a cohort of 16 fellowship-trained spine surgeons from 10 academic medical centers constructed a scoring system for the triage and prioritization of emergent and elective spine surgeries. Three separate rounds of videoconferencing and written correspondence were used to reach a final scoring system. Sixteen test cases were used to optimize the scoring system so that it could categorize cases as requiring emergent, urgent, high-priority elective, or low-priority elective scheduling. The devised scoring system included 8 independent components: neurologic status, underlying spine stability, presentation of a high-risk postoperative complication, patient medical comorbidities, expected hospital course, expected discharge disposition, facility resource limitations, and local disease burden. The resultant calculator was deployed as a freely available Web-based calculator (https://jhuspine3.shinyapps.io/SpineUrgencyCalculator/). We present the first quantitative urgency scoring system for the triage and prioritizing of spine surgery cases in resource-limited settings. We believe that our scoring system, although not all encompassing, has potential value as a guide for triaging spine surgical cases during the COVID pandemic and post-COVID period.


Characterization of visual pigments, oil droplets, lens and cornea in the whooping crane Grus americana.
Vision has been investigated in many species of birds, but few studies have considered the visual systems of large birds and the particular implications of large eyes and long-life spans on visual system capabilities. To address these issues we investigated the visual system of the whooping crane Grus americana (Gruiformes, Gruidae), which is one of only two North American crane species. It is a large, long-lived bird in which UV sensitivity might be reduced by chromatic aberration and entrance of UV radiation into the eye could be detrimental to retinal tissues. To investigate the whooping crane visual system we used microspectrophotometry to determine the absorbance spectra of retinal oil droplets and to investigate whether the ocular media (i.e. the lens and cornea) absorb UV radiation. In vitro expression and reconstitution was used to determine the absorbance spectra of rod and cone visual pigments. The rod visual pigments had wavelengths of peak absorbance (λmax) at 500 nm, whereas the cone visual pigment λmax values were determined to be 404 nm (SWS1), 450 nm (SWS2), 499 nm (RH2) and 561 nm (LWS), similar to other characterized bird visual pigment absorbance values. The oil droplet cut-off wavelength (λcut) values similarly fell within ranges recorded in other avian species: 576 nm (R-type), 522 nm (Y-type), 506 nm (P-type) and 448 nm (C-type). We confirm that G. americana has a violet-sensitive visual system; however, as a consequence of the λmax of the SWS1 visual pigment (404 nm), it might also have some UV sensitivity.


Effect of selected recovery conditions on performance of repeated bouts of intermittent cycling separated by 24 hours.
The purpose of this study was to examine the effects of active recovery (AR), massage (MR), and cold water immersion (CR) on performance of repeated bouts of high-intensity cycling separated by 24 hours. For each recovery condition, subjects were asked to take part in 2 intermittent cycling sessions; 18 minutes of varying work intervals performed in succession at a resistance of 80 g/kg body weight separated by 24 hours. One of four 15-minute recovery conditions immediately followed the first session and included: (a) AR, cycling at 30% Vo(2)max; (b) CR, immersion of legs in a 15 degrees C water bath; (c) MR, massage of the legs; and (d) control, seated rest. Only the control condition showed a significant decline in the total work completed between the first and second exercise sessions (108.1 +/- 5.4 kJ vs. 106.0 +/- 5.0 kJ, p < 0.05). Thus, AR, MR, and CR appeared to facilitate the recovery process between 2 high-intensity, intermittent exercise sessions separated by 24 hours.


Overcoming obstacles to late presentation for HIV infection in Europe.
The central goal of the HIV in Europe Initiative is to promote testing and treatment throughout Europe and Central Asia in order to decrease the number of people living with HIV presenting late for care. This article summarizes the results from the HIV in Europe 2009 Conference and the early results of the projects set up by the initiative, and discusses their implications for the future. In November 2009, 100 key stakeholders from 25 countries met in Stockholm at the HIV in Europe Conference. The focus was to address five key issues that contribute to the barriers to testing identified in 2007 at an innovative HIV conference. The conference discussed barriers to testing and other reasons for late presentation and outlined concrete recommendations to address the problem. An early result of the initiative has been stimulation of the process of reaching a consensus definition of what is meant by a 'late presenter', with this definition to be implemented at the European level. Steps are being taken to advocate for appropriate health policies and surveillance data related to HIV throughout Europe. Also, the initiative has set up projects related to the barriers to testing, i.e. criminalization law, stigmatization and lack of offering of testing for people presenting with certain indicator diseases. The final results of ongoing projects will be published and widely disseminated in 2010 and beyond. The HIV in Europe Initiative will continue to reinforce collaboration, advocacy and networking activities in the field throughout Europe.


Youth Sexual Health: Sexual Knowledge, Attitudes, and Behavior Among Students at a University in Turkey.
To determine sexual attitudes, behavior, and knowledge of Namik Kemal University (NKU) students about sexual health and sexually transmitted infections (STIs). A sample representing 10% of the undergraduate population of NKU in 2009-2010, was studied. Of 1,500 questionnaires distributed, 1,314 (87.6%) were filled out. The mean age of the respondents (52.9% male) was 20.07±1.75 years. The rate of students who had received sexual health education was 32.0%, and 15.3% had previously used a sexual health service. Eleven percent of the female students and 50.3% of the male students had had sexual intercourse. The average age of initial sexual intercourse was 16.83±2.07 years. Of the students who had had sexual intercourse, 46.6% reported that they did not use any contraception method. The most preferred method was condoms (37.6%). The rate of contraceptive use was 58.7% in sexually educated students and 43.9% in those not educated (p=.004). The most well-known STI was AIDS (96.5%), with sexually educated students giving higher rates of correct answers about STIs (p<.05). The students who had received sexual health education were more knowledgeable about vital consequences of STI's, even though it is not sufficient, than sexually active students. Awareness of safe sexual practices and changes in behavior, in particular, promoting condom use should be established in higher risk youths. Deficiencies in knowledge could be addressed by adding a sexual healthtraining component to the university curriculum, and unmet requirements could be met by reorganizing medico-social centers in universities.


Influence of probing force on the reproducibility of bleeding tendency measurements.
Three groups of seven patients were selected on the basis of about 50% loss of alveolar bone support in some part of the mouth. All patients received initial treatment consisting of plaque control and removal of subgingival deposits. In the first group of patients 95 interproximal pockets were measured three times by one examiner using the pressure probe employing a constant probing force of 0.75 N. Bleeding or lack of bleeding from a pocket was sought for up to 30 seconds after probing. Results show that a time interval of 15 minutes between reexamination procedures is enough to prevent an increased bleeding index caused by repeated probing. In the second group of patients, 98 interproximal pockets were measured three times with a Merritt-B probe (Hu-Friedy), each time by a different examiner. In the third group of patients, 102 interproximal pockets were measured three times with a pressure probe, each time by a different examiner. The same three examiners were involved. In both groups presence or absence of bleeding from a pocket was recorded. Results show a proportion of interexaminer agreement among three examiners of 0.40 with the Merritt-B probe and 0.71 with the pressure probe. It is concluded that a constant probing force of 0.75 N does contribute to more reproducible bleeding tendency measurements.