Hyperoxidation of Peroxiredoxins: Gain or Loss of Function?
In 2003, structural studies revealed that eukaryotic 2-Cys peroxiredoxins (Prx) have evolved to be sensitive to inactivation of their thioredoxin peroxidase activity by hyperoxidation (sulfinylation) of their peroxide-reacting catalytic cysteine. This was accompanied by the unexpected discovery, that the sulfinylation of this cysteine was reversible in vivo and the identification of a new enzyme, sulfiredoxin, that had apparently co-evolved specifically to reduce hyperoxidized 2-Cys Prx, restoring their peroxidase activity. Together, these findings have provided the impetus for multiple studies investigating the purpose of this reversible, Prx hyperoxidation. Recent Advances: It has been suggested that inhibition of the thioredoxin peroxidase activity by hyperoxidation can both promote and inhibit peroxide signal transduction, depending on the context. Prx hyperoxidation has also been proposed to protect cells against reactive oxygen species (ROS)-induced damage, by preserving reduced thioredoxin and/or by increasing non-peroxidase chaperone or signaling activities of Prx. Here, we will review the evidence in support of each of these proposed functions, in view of the in vivo contexts in which Prx hyperoxidation occurs, and the role of sulfiredoxin. Thus, we will attempt to explain the basis for seemingly contradictory roles for Prx hyperoxidation in redox signaling. We provide a rationale, based on modeling and experimental studies, for why Prx hyperoxidation should be considered a suitable, early biomarker for damaging levels of ROS. We discuss the implications that this has for the role of Prx in aging and the detection of hyperoxidized Prx as a conserved feature of circadian rhythms. Antioxid. Redox Signal. 28, 574-590.


Functional performance is associated with both knee extensor and flexor muscle strength in patients scheduled for total knee arthroplasty: A cross-sectional study.
To determine whether muscle strength in patients scheduled for total knee arthroplasty is: (i) strongly associated with both measured functional performance and patient-reported measures; (ii) more closely associated with functional performance when measured during concentric than during isometric contractions; and (iii) more strongly related to the 30-s chair stand test than to the timed-up-and-go and walking measures. Cross-sectional-study. Fifty-nine patients (36 women, 23 men), mean age 70.4 years. Associations between muscle strength, measured functional performance, and patient-reported measures were calculated. Both knee extensor and knee flexor strength were associated with performance-based measures. In general, concentric knee flexor muscle strength was more strongly associated with functional performance than was isometric knee flexor strength. Concentric and isometric knee extensor strength were of equal importance. The 30-s chair stand test was better than the timed-up-and-go and the walking tests at determining muscle strength. Future rehabilitation programmes should include both the knee extensor muscles and the knee flexor muscles in order to improve functional performance. The 30-s chair stand test is a valid and clinical relevant proxy measure of knee extensor and knee flexor muscle strength.


P gene mutations associated with oculocutaneous albinism type II (OCA2).
Oculocutaneous albinism type II (OCA2) is the most common form of albinism in humans. OCA2 has been previously associated with mutations of the P gene, the human homologue to the murine pink-eyed dilution gene. The P gene encodes a 110 kDa protein containing 12 potential membrane spanning domains and is associated with melanosomal membranes. The specific function of the P protein is currently unknown but is thought to be involved in tyrosinase processing and transport. We report nine novel mutations in the P gene associated with OCA2. These include two missense mutations, c.1938A>C (p.Ile646Val) and c.1556T>C (p.Val519Ala); one nonsense mutation c.612G>A (p.Trp204X); five frameshift mutations: c.2372_2373delTC, c.1555delG, c.1938_1939insC, c.2050delT, and c.1045_1046delAT; and a splice site mutation c.1951+1G>A. We also report 12 novel polymorphisms including one amino acid substitution, c.2365_2366GC>CA (p.Ala789Glu). At present, there is no functional assay to determine if a mutation is truly pathogenic. The presence of numerous polymorphisms of the P gene in the coding region, several of which result in amino acid substitutions, makes molecular diagnosis problematic. To ensure accurate molecular diagnosis, further mutational analysis will be necessary to produce a comprehensive list of mutations associated with OCA2. This information will also help define the critical functional domains of the P protein. Mutations associated with OCA2 can be found in the Albinism Database (http://albinismdb.med.umn.edu).


Matrix Fabry-Perot resonance mechanism in high-contrast gratings.
We present a simple analytic formalism to explain the unique resonance phenomenon in subwavelength high-contrast gratings (HCG). We show that the resonances are due to strong coupling between two surface-normal waveguide array modes resulting from abrupt and large index contrast. Simple expression for HCG quality factor is derived that agrees with spectral-fitting approaches reported in literature.


Reversal by phenytoin of carbamazepine-induced water intoxication: a pharmacokinetic interaction.
The hypothesis that phenytoin may antagonise the antidiuretic effect of carbamazepine has been examined by comparing the free water clearance response to a standard water load in 36 patients stabilised on different drug regimes. The diuretic response to the water load was significantly greater in patients receiving chronic treatment with carbamazepine and phenytoin in combination than in matched control subjects receiving carbamazepine as a single drug. Acute administration of phenytoin (1,100 mg), however, had no significant influence on carbamazepine-induced antidiuresis. Evidence is presented that reversal of the antidiuretic effect of carbamazepine by chronic phenytoin administration is secondary to a marked reduction of the serum carbamazepine concentration during combined therapy. These results suggest that the risk of developing water intoxication is greater in patients receiving carbamazepine alone than in those receiving phenytoin in combination. Since the antidiuretic effect is correlated with the serum carbamazepine concentration rather than with the prescribed daily dose, monitoring the serum level of the drug is likely to provide the best rational approach to the prevention of excessive water retention.


Non-neoplastic pulmonary multinucleate giant cells. Comparative study of multinucleate giant cells in porcine dermatosis vegetans and other polykaryocytic conditions.
Pulmonary multinucleate giant cells (MGCs) in porcine dermatosis vegetans (DV) were compared with virus-induced and inflammatory-associated MGCs, with particular attention to mechanisms involved in the formation of such cells. Ultrastructurally, pigs with DV showed numerous pulmonary MGCs with transversely-sectioned interdigitating microvillous structures. Microvilli were observed closely apposed to type-II pneumocytes and with morphological characteristics of membrane bridge formation. Rabbits infected with Mycobacterium bovis exhibited multiple pulmonary granulomas with prominent MGC formation 14 days after infection. Close apposition between MGCs and small macrophages and membrane bridge formation were demonstrated ultrastructurally. Pulmonary changes in calves infected with bovine respiratory syncytial virus (BRSV) included lobular consolidation and intense infiltration of inflammatory cells into the bronchial, bronchiolar and alveolar walls. Proliferation of bronchiolo-alveolar epithelial cells was prominent, but syncytiae were only occasionally demonstrated. This study of the morphological characteristics of MGCs is related to the present knowledge of mechanisms involved in membrane fusion and allows for an interpretation of an active fusion process involved in the formation of MGCs in porcine DV.


The SWI/SNF-like BAF complex is essential for early B cell development.
During the process of B cell development, transcription factors, such as E2A and Ebf1, have been known to play key roles. Although transcription factors and chromatin regulators work in concert to direct the expression of B lineage-specific genes, little is known about the involvement of regulators for chromatin structure during B lymphopoiesis. In this article, we show that deletion of Srg3/mBaf155, a scaffold subunit of the SWI/SNF-like BAF complex, in the hematopoietic lineage caused defects at both the common lymphoid progenitor stage and the transition from pre-pro-B to early pro-B cells due to failures in the expression of B lineage-specific genes, such as Ebf1 and Il7ra, and their downstream target genes. Moreover, mice that were deficient in the expression of Brg1, a subunit of the complex with ATPase activity, also showed defects in early B cell development. We also found that the expression of Ebf1 and Il7ra is directly regulated by the SWI/SNF-like BAF complex. Thus, our results suggest that the SWI/SNF-like BAF complex facilitates early B cell development by regulating the expression of B lineage-specific genes.


Before and After: What Has Changed in the Attitude of Turkish Society Regarding Face Transplantations?
Face transplantation is one of the most popular and controversial subjects of plastic surgery today. Although there are various surveys on the subject, there is no study comparing the past and the present social viewpoint and behavioral preferences for face transplantations across the world. In this study, we aimed to investigate the changes in the views of the Turkish society with respect to face transplantation from past to present. For this purpose, 1000 volunteer participants were questioned in terms of demographics and their perspective and preferences on organ and face transplantation. The results of the study were compared with the past data, and based on the results, the level of consciousness and awareness of the Turkish society about the subject has increased; the rate risk taking for immunosuppression has decreased, and instead, the rate of having an undecided stance has increased, and this attitude continues even if the risk is resolved. With these results, we can conclude that the greatest handicap for face transplantation in the Turkish society today is immunosuppression and the associated risks. We believe that new drug protocols and monitoring of patient outcomes for longer periods as well as more extensive clinical applications may be beneficial in addressing this issue.


Immunogenically fit subunit vaccine components via epitope discovery from natural peptide libraries.
Antigenic peptides that bind pathogen-specific Abs are a potential source of subunit vaccine components. To be effective the peptides must be immunogenically fit: when used as immunogens they must elicit Abs that cross-react with native intact pathogen. In this study, antigenic peptides obtained from phage display libraries through epitope discovery were systematically examined for immunogenic fitness. Peptides selected from random peptide libraries, in which the phage-displayed peptides are encoded by synthetic degenerate oligonucleotides, had marginal immunogenic fitness. In contrast, 50% of the peptides selected from a natural peptide library, in which phage display segments of actual pathogen polypeptides, proved very successful. Epitope discovery from natural peptide libraries is a promising route to subunit vaccines.


Computed tomography for patient management in coronary artery disease.
Computed tomography (CT) represents the optimum theoretical approach to x-ray imaging. This conclusion results from an awareness of CT's capacity to solve the fundamental limitation of all forms of x-ray imaging--the superimposition of anatomic structures. Because CT is potentially a fully three-dimensional method, this problem is addressed in a manner not subject to the risks, complications, and technical limitations of selective angiography, subtraction angiography, tomography, and other techniques. Ultrafast CT provides a broad spectrum of useful quantitative cardiac data during one minimally invasive procedure. Therefore, the optimism for cine CT imaging of the heart is well founded. Apart from the demonstration of anatomic structures in any reconstructed plane and in movie format, without the need for any form of electrocardiographic gating, this new generation of millisecond CT scanners offers a unique potential for measuring myocardial mechanics and perfusion. Myocardial integrity can be evaluated by measuring myocardial wall thickening, which is a sensitive indicator of blood flow. It can also be assessed by time-density changes derived from analyzing the passage of contrast medium through thin slices of myocardium. Feasibility studies have demonstrated that this should be possible in the clinical setting by use of fast CT scanning. The radiation exposure to the patient with cine CT is low and comparable to, or less than, that of conventional CT and 10-15% of that received during angiocardiography; therefore, it is not a practical limitation. Further clinical and research studies are needed to determine the future role of this exciting new modality in the diagnosis and management of patients with heart disease.


Validation of a PCR for diagnosis of typhoid fever and salmonellosis by amplification of the hilA gene in clinical samples from Colombian patients.
Validation of a PCR test to detect hilA gene sequences of Salmonella spp. was performed in blood and faeces samples from typhoid fever and salmonellosis patients. Sensitivity (S), specificity (SP), positive predictive value (PPV) and negative predictive value (NPV) of the PCR in blood samples were performed by testing: 37 patients with clinical diagnosis of typhoid fever, 34 of them confirmed by isolation of S. Typhi from blood cultures; 35 patients infected with other pathogens corroborated by blood culture (Klebsiella pneumoniae, 9; Serratia marcescens, 5; Escherichia coli, 4; Pseudomonas aeruginosa, 9; Providencia alcalifaciens, 4 and Enterobacter cloacae, 4) and blood samples from 150 healthy volunteers. To evaluate S, SP, PPV and NPV of the PCR in faeces samples we studied: 34 patients with enteritis due Salmonella spp. (S. Typhimurium, 21; S. Enteritidis, 9; S. Choleraesuis, 3 and S. Agona, 1); faeces samples from 35 patients with enteric infection due to Shigella sonnei (8), Shigella flexneri (10), enteropathogenic E. coli (12), Aeromonas hydrophila (5) and faeces samples from 150 healthy volunteers. The S, SP, PPV and NPV of the PCR in blood samples were all 100 %. PCR detected three patients with clinical diagnosis of typhoid fever and negative blood cultures. In faeces samples, S was 97 %, SP 100 %, PPV 100 % and NPV 99 %. The lowest number of c.f.u. ml(-1) detected by PCR in blood samples was 1 x 10(1) and in faeces samples 4 x 10(2).


miR-511-3p protects against cockroach allergen-induced lung inflammation by antagonizing CCL2.
miR-511-3p, encoded by CD206/Mrc1, was demonstrated to reduce allergic inflammation and promote alternative (M2) macrophage polarization. Here, we sought to elucidate the fundamental mechanism by which miR-511-3p attenuates allergic inflammation and promotes macrophage polarization. Compared with WT mice, the allergen-challenged Mrc1-/- mice showed increased airway hyperresponsiveness (AHR) and inflammation. However, this increased AHR and inflammation were significantly attenuated when these mice were pretransduced with adeno-associated virus-miR-511-3p (AAV-miR-511-3p). Gene expression profiling of macrophages identified Ccl2 as one of the major genes that was highly expressed in M2 macrophages but antagonized by miR-511-3p. The interaction between miR-511-3p and Ccl2 was confirmed by in silico analysis and mRNA-miR pulldown assay. Further evidence for the inhibition of Ccl2 by miR-511-3p was given by reduced levels of Ccl2 in supernatants of miR-511-3p-transduced macrophages and in bronchoalveolar lavage fluids of AAV-miR-511-3p-infected Mrc1-/- mice. Mechanistically, we demonstrated that Ccl2 promotes M1 macrophage polarization by activating RhoA signaling through Ccr2. The interaction between Ccr2 and RhoA was also supported by coimmunoprecipitation assay. Importantly, inhibition of RhoA signaling suppressed cockroach allergen-induced AHR and lung inflammation. These findings suggest a potentially novel mechanism by which miR-511-3p regulates allergic inflammation and macrophage polarization by targeting Ccl2 and its downstream Ccr2/RhoA axis.


Sympatric cryptic species in the crinoid genus Cenolia (Echinodermata: Crinoidea: Comasteridae) delineated by sequence and microsatellite markers.
The marine species of the southern coast of Australia have not been well studied with regard to molecular connectivity. Cryptic species are expected to be prevalent on this coastline. Here, we investigate the crinoid genus Cenolia (Echinodermata: Crinoidea: Comasteridae) using molecular methods to elucidate cryptic species and phylogenetic relationships. The genus Cenolia dominates the southern Australian crinoid fauna in shallow waters. Few studies have examined crinoids for cryptic species at a molecular level and these have been predominantly based on mitochondrial data. We employ the nuclear markers 28S rRNA and ITS-2 in addition to the mitochondrial COI. Six divergent mitochondrial clades were identified. Gene flow between confirmed clades was subsequently examined by the use of six novel microsatellite markers, showing that sympatric taxa with low mtDNA divergences (1.7% K2P) were not interbreeding in the wild. The type specimens of Cenolia benhami and C. spanoschistum were examined, as well as all six divergent clades. Morphological characters dividing taxa were refined. Due to comb pinnule morphology, the New Zealand species benhami was determined to belong to the genus Oxycomanthus (nov. comb.). Three new species of Cenolia (including the Australian "benhami") require description.


Aliphatic Chain Modification of Collagen Type I: Development of Elastomeric, Compliant, and Suturable Scaffolds.
Collagen type I is one of the most suitable natural biomaterials for constructing tissue-engineering scaffolds. Despite their biocompositional similarities to physiological tissues, these scaffolds lack host specific and matching mechanical properties. While it is possible to enhance their stiffness by cross-linking, it often compromises their abilities to expand or strain under minimal stress, that is, compliance (inverse of stiffness). Here, we report a simple, inexpensive, cross-linking- and elastin-free collagen-based material composition for developing elastomeric scaffolds that are highly compliant, soft yet strong, and suturable, therefore, clinically attractive. Our strategy utilizes room-temperature modification of collagen type I scaffolds with linear aliphatic chains of various lengths (C7-C18). In particular, dodecenylsuccinic anhydride (size: C12, DDSA) modified scaffolds elongated up to 400% of its initial length compared to only ∼20% for collagen-control within the applied tensile stress of 0.2 MPa without breaking. Furthermore, the suture retention strength value increased to 60 g-force from 30 g-force for collagen control. We confirmed that the C12-modified material remained structurally stable at the physiological temperature (37 °C) with a tan δ value of ∼0.3, similar to collagen control; however, tan δ increased sharply for C12-modified collagen above 42 °C, compared to 59 °C for collagen control. To understand the mechanism of hyperextensibility, we studied the morphology of the resultant material by transmission electron microscopy (TEM), which showed an altered microstructure of C12-modified collagen scaffolds. While the partially C12-modified sample had a mixture of typical collagen type I triple helix and diffused gelatinized random coil-like configuration, the fully modified samples showed thick wrinkled and entangled ribbon-like microstructures, which was different than that of thermally denatured gelatin. We further confirmed that the resultant material allowed cell growth in vitro and in vivo in a subcutaneous mouse model.


Arterial resection at the time of pancreatectomy for cancer.
Tumor-induced arterial abutment/encasement has been traditionally a contraindication to surgery in patients with localized pancreatic cancer (PC). One recent meta-analysis reported greater mortality rates in this setting. We report herein a series of planned arterial resections in carefully selected patients who responded favorably to combined modality therapy for localized PC. We reviewed all patients with PC and arterial encasement treated between May 2011 and September 2013; all patients received an extensive course of neoadjuvant therapy before surgery. Of 15 patients taken to surgery, 2 had peritoneal disease at laparoscopy, and therefore, laparotomy was not performed. Pancreatectomy (pancreaticoduodenectomy, 3; distal, 8; central pancreatectomy, 1; total, 1) was performed in the remaining 13, 10 of whom required arterial resection. The most common operation was an Appleby procedure. Of 10 patients who underwent combined pancreatectomy and arterial resection, their median age was 62 years (range, 33-75), median operative time was 7.5 hours, and median blood loss was 725 mL. Complications occurred in 3 of 15 patients with no perioperative mortality. Median duration of hospital stay was 9 days (range, 5-19). An R0 resection was achieved in 11 (85%) of 13 patients. At a median follow-up of 21 months, 8 of these 13 resected patients (62%) are alive without disease. Planned arterial resection at the time of pancreatectomy can be performed with acceptable morbidity and mortality; patient selection and induction therapy are likely critically important variables that seem to impact patient outcome. Those patients with stable or responding disease after induction therapy represent the subset of patients with potentially favorable tumor biology in whom extended resections may enhance survival duration.


Vulnerable Subjects: Why Does Informed Consent Matter?
This special issue of the Journal Law, Medicine & Ethics takes up the concern of informed consent, particularly in times of controversy. The dominant moral dilemmas that frame traditional bioethical concerns address medical experimentation on vulnerable subjects; physicians assisting their patients in suicide or euthanasia; scarce resource allocation and medical futility; human trials to develop drugs; organ and tissue donation; cloning; xenotransplantation; abortion; human enhancement; mandatory vaccination; and much more. The term "bioethics" provides a lens, language, and guideposts to the study of medical ethics. It is worth noting, however, that medical experimentation is neither new nor exclusive to one country. Authors in this issue address thorny subjects that span borders and patients: from matters dealing with children and vaccination to the language and perception of consent.


Measuring the impact of health insurance with a correction for selection bias--a case study of Ecuador.
This article develops and uses methodologies to evaluate the impact of publicly-financed health insurance programmes on the use of health care. Using univariate and bivariate probit estimation techniques, the study tests and corrects for endogeneity resulting from selection bias. Potential endogeneity arises from the choice to be insured, eligibility for insurance, and differences in individuals' health status. The setting for the study is the country of Ecuador. The General Health Insurance (GHI) programme, which primarily covers workers in the formal sector of the economy, is found to have a strong positive association with the use of curative health care after correcting for selection bias, but no significant effect on the use of preventive care. Individuals with severe illnesses who are eligible for GHI have a preference for private health care, and self-select out of the GHI programme. The Seguro Campesino Social (SSC) programme, directed at farming populations, has positive but insignificant associations with both curative and preventive care.


Lubrication, adsorption, and rheology of aqueous polysaccharide solutions.
Aqueous lubrication is currently at the forefront of tribological research due to the desire to learn and potentially mimic how nature lubricates biotribological contacts. We focus here on understanding the lubrication properties of naturally occurring polysaccharides in aqueous solution using a combination of tribology, adsorption, and rheology. The polysaccharides include pectin, xanthan gum, gellan, and locus bean gum that are all widely used in food and nonfood applications. They form rheologically complex fluids in aqueous solution that are both shear thinning and elastic, and their normal stress differences at high shear rates are found to be characteristic of semiflexible/rigid molecules. Lubrication is studied using a ball-on-disk tribometer with hydrophobic elastomer surfaces, mimicking biotribological contacts, and the friction coefficient is measured as a function of speed across the boundary, mixed, and hydrodynamic lubrication regimes. The hydrodynamic regime, where the friction coefficient increases with increasing lubricant entrainment speed, is found to depend on the viscosity of the polysaccharide solutions at shear rates of around 10(4) s(-1). The boundary regime, which occurs at the lowest entrainment speeds, depends on the adsorption of polymer to the substrate. In this regime, the friction coefficient for a rough substrate (400 nm rms roughness) is dependent on the dry mass of polymer adsorbed to the surface (obtained from surface plasmon resonance), while for a smooth substrate (10 nm rms roughness) the friction coefficient is strongly dependent on the hydrated wet mass of adsorbed polymer (obtained from quartz crystal microbalance, QCM-D). The mixed regime is dependent on both the adsorbed film properties and lubricant's viscosity at high shear rates. In addition, the entrainment speed where the friction coefficient is a minimum, which corresponds to the transition between the hydrodynamic and mixed regime, correlates linearly with the ratio of the wet mass and viscosity at ∼10(4) s(-1) for the smooth surface. These findings are independent of the different polysaccharides used in the study and their different viscoelastic flow properties.


Influence of mandibular morphology on the hyoid bone in atypical deglutition: a correlational study.
evaluate the possible correlation with the radiographic position of the hyoid bone and mandibular angle in lateral radiographs of children with atypical deglutition. This was an observational study using cephalometric analysis of lateral teleradiographs for the distances of H-MP (hyoid to mandibular plane). Spearman's correlation analysis was performed with MA (mandibular angle) in two groups: the experimental group with atypical deglutition and the control group normal deglutition. Both groups included subjects in mixed dentition stage. there was a significant moderate negative correlation between MA (mandibular angle) and hyoid bone (H-MP) in the normal group (R = -0.406, p = 0.021). However, there was no significant correlation between the MA and H-MP (R = 0.029, p = 0.83) in the group with atypical deglutition. there is a moderate negative correlation between the position of the hyoid bone and mandibular angle in the group of normal swallowing and there is no correlation between variables H-MP and MA in the group of atypical swallowing.


Perioperative management of obstructive sleep apnea.
Obstructive sleep apnea (OSA) is the most common breathing disorder, with a high prevalence in both the general and surgical populations. OSA is frequently undiagnosed, and the initial recognition often occurs during medical evaluation undertaken to prepare for surgery. Adverse respiratory and cardiovascular outcomes are associated with OSA in the perioperative period; therefore, it is imperative to identify and treat patients at high risk for the disease. In this review, we discuss the epidemiology of OSA in the surgical population and examine the available data on perioperative outcomes. We also review the identification of high-risk patients using clinical screening tools and suggest intraoperative and postoperative treatment regimens. Additionally, the role of continuous positive airway pressure in perioperative management of OSA and a brief discussion of ambulatory surgery in patients with OSA is provided. Finally, an algorithm to guide perioperative management is suggested.


Action planning and predictive coding when speaking.
Across the animal kingdom, sensations resulting from an animal's own actions are processed differently from sensations resulting from external sources, with self-generated sensations being suppressed. A forward model has been proposed to explain this process across sensorimotor domains. During vocalization, reduced processing of one's own speech is believed to result from a comparison of speech sounds to corollary discharges of intended speech production generated from efference copies of commands to speak. Until now, anatomical and functional evidence validating this model in humans has been indirect. Using EEG with anatomical MRI to facilitate source localization, we demonstrate that inferior frontal gyrus activity during the 300ms before speaking was associated with suppressed processing of speech sounds in auditory cortex around 100ms after speech onset (N1). These findings indicate that an efference copy from speech areas in prefrontal cortex is transmitted to auditory cortex, where it is used to suppress processing of anticipated speech sounds. About 100ms after N1, a subsequent auditory cortical component (P2) was not suppressed during talking. The combined N1 and P2 effects suggest that although sensory processing is suppressed as reflected in N1, perceptual gaps may be filled as reflected in the lack of P2 suppression, explaining the discrepancy between sensory suppression and preserved sensory experiences. These findings, coupled with the coherence between relevant brain regions before and during speech, provide new mechanistic understanding of the complex interactions between action planning and sensory processing that provide for differentiated tagging and monitoring of one's own speech, processes disrupted in neuropsychiatric disorders.


The central nervous system is a viral reservoir in simian immunodeficiency virus--infected macaques on combined antiretroviral therapy: a model for human immunodeficiency virus patients on highly active antiretroviral therapy.
This study used a simian immunodeficiency virus (SIV)-macaque model to determine whether virus persists in the central nervous system (CNS) of human immunodeficiency virus (HIV)-infected individuals in which plasma viral load has been suppressed by highly active antiretroviral therapy. SIV-infected macaques were treated with two reverse transcriptase inhibitors: PMPA (q- R-(2-phosphonomethoxypropyl)adenine)which does not cross the blood-brain barrier, and FTC (beta-2('),3(')-dideoxy-3 thia-5-fluorocytidine), which does. Viral DNA and RNA were quantitated in the brain after 6 months of suppression of virus replication in blood and cerebrospinal fluid (CSF). Viral DNA was detected in brain from all macaques, including those in which peripheral viral replication had been suppressed either by antiretroviral therapy or host immune responses. Significant neurological lesions were observed only in one untreated macaque that had active virus replication in the CNS. Expression of the inflammatory markers, major histocmopatibility complex (MHC) II and CD68 was significantly lower in macaques treated with PMPA/FTC. Thus, although antiretroviral treatment may suppress virus replication in the periphery and the brain and reduce CNS inflammation, viral DNA persists in the brain despite treatment. This suggests that the brain may serve as a long-term viral reservoir in HIV-infected individuals treated with antiretroviral drugs that suppress virus replication.


Characterization of binding of LARP6 to the 5' stem-loop of collagen mRNAs: implications for synthesis of type I collagen.
Type I collagen is composed of 2 polypeptides, α1(I) and α2(I), which fold into triple helix. Collagen α1(I) and α2(I) mRNAs have a conserved stem-loop structure in their 5' UTRs, the 5'SL. LARP6 binds the 5'SL to regulate type I collagen expression. We show that 5 nucleotides within the single stranded regions of 5'SL contribute to the high affinity of LARP6 binding. Mutation of individual nucleotides abolishes the binding in gel mobility shift assay. LARP6 binding to 5'SL of collagen α2(I) mRNA is more stable than the binding to 5'SL of α1(I) mRNA, although the equilibrium binding constants are similar. The more stable binding to α2(I) mRNA may favor synthesis of the heterotrimeric type I collagen. LARP6 needs 2 domains to contact 5'SL, the La domain and the RRM. T133 in the La domain is critical for folding of the protein, while loop 3 in the RRM is critical for binding 5'SL. Loop 3 is also involved in the interaction of LARP6 and protein translocation channel SEC61. This interaction is essential for type I collagen synthesis, because LARP6 mutant which binds 5'SL but which does not interact with SEC61, suppresses collagen synthesis in a dominant negative manner. We postulate that LARP6 directly targets collagen mRNAs to the SEC61 translocons to facilitate coordinated translation of the 2 collagen mRNAs. The unique sequences of LARP6 identified in this work may have evolved to enable its role in type I collagen biosynthesis.


Acute effects of acetazolamide in hyperkalemic periodic paralysis.
We studied the effect of acetazolamide on plasma potassium in normals and in two patients with hyperkalemic periodic paralysis. Administration of acetazolamide for 48 hours lowered mean plasma potassium in normals from 4.01 to 3.56 mEq per liter (p less than 0.001) and in the patients from 4.55 to 4.00 mEq per liter (p less than 0.001). This kaliopenic effect of acetazolamide may account for its therapeutic action in hyperkalemic periodic paralysis.


Endocannabinoid Modulation of Microglial Phenotypes in Neuropathology.
Microglia, the resident immune cells of the central nervous system, mediate brain homeostasis by controlling neuronal proliferation/differentiation and synaptic activity. In response to external signals from neuropathological conditions, homeostatic (M0) microglia can adopt one of two activation states: the classical (M1) activation state, which secretes mediators of the proinflammatory response, and the alternative (M2) activation state, which presumably mediates the resolution of neuroinflammation and tissue repair/remodeling. Since chronic inflammatory activation of microglia is correlated with several neurodegenerative diseases, functional modulation of microglial phenotypes has been considered as a potential therapeutic strategy. The endocannabinoid (eCB) system, composed of cannabinoid receptors and ligands and their metabolic/biosynthetic enzymes, has been shown to activate anti-inflammatory signaling pathways that modulate immune cell functions. Growing evidence has demonstrated that endogenous, synthetic, and plant-derived eCB agonists possess therapeutic effects on several neuropathologies; however, the molecular mechanisms that mediate the anti-inflammatory effects have not yet been identified. Over the last decade, it has been revealed that the eCB system modulates microglial activation and population. In this review, we thoroughly examine recent studies on microglial phenotype modulation by eCB in neuroinflammatory and neurodegenerative disease conditions. We hypothesize that cannabinoid 2 receptor (CB2R) signaling shifts the balance of expression between neuroinflammatory (M1-type) genes, neuroprotective (M2-type) genes, and homeostatic (M0-type) genes toward the latter two gene expressions, by which microglia acquire therapeutic functionality.


A subset of human plasmacytoid dendritic cells expresses CD8α upon exposure to herpes simplex virus type 1.
Classical and plasmacytoid dendritic cells (DC) play important roles in the defense against murine and human infections with herpes simplex virus (HSV). So far, CD8α expression has only been reported for murine DC. CD8α(+) DC have prominent cross-presenting activities, which are enhanced by murine CD8α(+) PDC. The human orthologue of murine CD8α(+) DC, the CD141 (BDCA3)(+) DC, mainly cross-present after TLR3 ligation. We report here the serendipitous finding that a subset of human PDC upregulates CD8α upon HSV-1 stimulation, as shown by gene array and flow cytometry analyses. CD8α, not CD8ß, was expressed upon exposure. Markers of activation, migration, and costimulation were upregulated on CD8α-expressing human PDC. In these cells, increased cytokine and chemokine levels were detected that enhance development and function of T, B, and NK cells, and recruit immature DC, monocytes, and Th1 cells, respectively. Altogether, human CD8α(+) PDC exhibit a highly activated phenotype and appear to recruit other immune cells to the site of inflammation. Further studies will show whether CD8α-expressing PDC contribute to antigen cross-presentation, which may be important for immune defenses against HSV infections in vitro and in vivo.


Standardized measurement of recovery from nonspecific back pain.
To propose standardized, patient-centered measures of recovery from nonspecific low back pain (LBP) in research, underpinned by an empirically derived concept of recovery and informed by expert opinion. Synthesis of literature reviews and expert panel opinion. Primary care centers for the management of nonspecific LBP. Persons with nonspecific LBP. Conservative treatments for nonspecific LBP. Three phases of research were conducted. First, qualitative research that explored patients' perspectives of recovery from nonspecific LBP was reviewed. Second, measures of recovery used in LBP clinical trials during the past decade were investigated in a systematic review. Third, opinion was sought from an expert panel of clinicians and researchers about how to measure recovery from nonspecific LBP, in a workshop at the 10th International Forum for Primary Care Research in Low Back Pain. An empirically derived and patient-centered concept of recovery from nonspecific LBP was developed from the qualitative research phase. The systematic review conducted in the second study phase revealed that researchers have used vastly heterogeneous measures of LBP recovery in clinical trials during the past decade. Finally, the key conclusions of the LBP Forum workshop were (1) that appropriate patient-centered instruments to measure recovery include global measures and patient-specific measures; and (2) that the benefits of implementing the same recovery measures for acute and chronic LBP outweigh the disadvantages of using different measures. The results were synthesized to inform our recommendation that researchers consider adopting 2 instruments as standardized measures of recovery from nonspecific LBP, as an adjunct to the existing core set of LBP outcome measures. These instruments are an 11-point Global Back Recovery Scale, for a simple measure of global recovery, and the Patient-Generated Index of Quality of Life-Back Pain, to evaluate specific relevant dimensions of recovery. This recommendation has majority endorsement by members of the Australian National Health and Medical Research Council Acute Low Back Pain Review Group.


Baicalein increases cisplatin sensitivity of A549 lung adenocarcinoma cells via PI3K/Akt/NF-κB pathway.
Baicalein, a bioactive flavonoid, exhibits anti-inflammatory and anti-cancer activities. However, few studies reported the interaction of baicalein with chemotherapeutic agents. Our study showed that baicalein significantly enhanced the chemosensitivity of cisplatin (CDDP) in vivo and in vitro. We found that A549/CDDP (resistant to CDDP) cells not only acquired epithelial-mesenchymal transition (EMT) phenotype, but also showed increased NF-κB activity compared with A549 cells (sensitive to CDDP). Our study further demonstrated that PI3K/Akt/NF-κB pathway controlled CDDP resistance via EMT and NF-κB-mediated apoptosis. Baicalein significantly suppressed the PI3K/Akt/NF-κB pathway, leading to conversion of EMT to mesenchymal-epithelial transition (MET, the reciprocal mesenchymal to epithelial transition), and inhibition of NF-κB-mediated antiapoptotic proteins in A549/CDDP cells. In conclusion, our study demonstrated that baicalein reversed the resistance of human A549 lung adenocarcinoma cells to cisplatin by inhibiting EMT and attenuating apoptosis via PI3K/Akt/NF-κB pathway.


Red Sea corals under Artificial Light Pollution at Night (ALAN) undergo oxidative stress and photosynthetic impairment.
Coral reefs represent the most diverse marine ecosystem on the planet, yet they are undergoing an unprecedented decline due to a combination of increasing global and local stressors. Despite the wealth of research investigating these stressors, Artificial Light Pollution at Night (ALAN) or "ecological light pollution" represents an emerging threat that has received little attention in the context of coral reefs, despite the potential of disrupting the chronobiology, physiology, behavior, and other biological processes of coral reef organisms. Scleractinian corals, the framework builders of coral reefs, depend on lunar illumination cues to synchronize their biological rhythms such as behavior, reproduction and physiology. While, light pollution (POL) may mask and lead de-synchronization of these biological rhythms process. To reveal if ALAN impacts coral physiology, we have studied two coral species, Acropora eurystoma and Pocillopora damicornis, from the Gulf of Eilat/Aqaba, Red Sea, which is undergoing urban development that has led to severe POL at night. Our two experimental design data revealed that corals exposed to ALAN face an oxidative stress condition, show lower photosynthesis performances measured by electron transport rate (ETR), as well as changes in chlorophyll and algae density parameters. Testing different lights such as Blue LED and White LED spectrum showed more extreme impact in comparison to Yellow LEDs on coral physiology. The finding of this work sheds light on the emerging threat of POL and the impacts on the biology and ecology of Scleractinian corals, and will help to formulate specific management implementations to mitigate its potentially harmful impacts.


Collaboration rules.
Corporate leaders seeking to boost growth, learning, and innovation may find the answer in a surprising place: the Linux open-source software community. Linux is developed by an essentially volunteer, self-organizing community of thousands of programmers. Most leaders would sell their grandmothers for workforces that collaborate as efficiently, frictionlessly, and creatively as the self-styled Linux hackers. But Linux is software, and software is hardly a model for mainstream business. The authors have, nonetheless, found surprising parallels between the anarchistic, caffeinated, hirsute world of Linux hackers and the disciplined, tea-sipping, clean-cut world of Toyota engineering. Specifically, Toyota and Linux operate by rules that blend the self-organizing advantages of markets with the low transaction costs of hierarchies. In place of markets' cash and contracts and hierarchies' authority are rules about how individuals and groups work together (with rigorous discipline); how they communicate (widely and with granularity); and how leaders guide them toward a common goal (through example). Those rules, augmented by simple communication technologies and a lack of legal barriers to sharing information, create rich common knowledge, the ability to organize teams modularly, extraordinary motivation, and high levels of trust, which radically lowers transaction costs. Low transaction costs, in turn, make it profitable for organizations to perform more and smaller transactions--and so increase the pace and flexibility typical of high-performance organizations. Once the system achieves critical mass, it feeds on itself. The larger the system, the more broadly shared the knowledge, language, and work style. The greater individuals' reputational capital, the louder the applause and the stronger the motivation. The success of Linux is evidence of the power of that virtuous circle. Toyota's success is evidence that it is also powerful in conventional companies.


Current and future molecular diagnostics for ocular infectious diseases.
Confirmation of ocular infections can pose great challenges to the clinician. A fundamental limitation is the small amounts of specimen that can be obtained from the eye. Molecular diagnostics can circumvent this limitation and have been shown to be more sensitive than conventional culture. The purpose of this review is to describe new molecular methods and to discuss the applications of next-generation sequencing-based approaches in the diagnosis of ocular infections. Efforts have focused on improving the sensitivity of pathogen detection using molecular methods. This review describes a new molecular target for Toxoplasma gondii-directed polymerase chain reaction assays. Molecular diagnostics for Chlamydia trachomatis and Acanthamoeba species are also discussed. Finally, we describe a hypothesis-free approach, metagenomic deep sequencing, which can detect DNA and RNA pathogens from a single specimen in one test. In some cases, this method can provide the geographic location and timing of the infection. Pathogen-directed PCRs have been powerful tools in the diagnosis of ocular infections for over 20 years. The use of next-generation sequencing-based approaches, when available, will further improve sensitivity of detection with the potential to improve patient care.


Teaching reciprocal imitation skills to young children with autism using a naturalistic behavioral approach: effects on language, pretend play, and joint attention.
Children with autism exhibit significant deficits in imitation skills which impede the acquisition of more complex behaviors and socialization, and are thus an important focus of early intervention programs for children with autism. This study used a multiple-baseline design across five young children with autism to assess the benefit of a naturalistic behavioral technique for teaching object imitation. Participants increased their imitation skills and generalized these skills to novel environments. In addition, participants exhibited increases in other social-communicative behaviors, including language, pretend play, and joint attention. These results provide support for the effectiveness of a naturalistic behavioral intervention for teaching imitation and offer a new and potentially important treatment option for young children who exhibit deficits in social-communicative behaviors.


Homing in on acute graft vs. host disease: tissue-specific T regulatory and Th17 cells.
Acute graft vs. host disease (aGVHD) is a major limitation of hematopoietic stem cell transplantation (HSCT), and it causes significant morbidity and mortality for this patient population. This immune-mediated injury occurs unpredictably and is caused by donor-derived T cells reacting to recipient alloantigens. Although donor Th1 cells play a critical role in aGVHD generation, numerous arms of both the innate and the adaptive immune systems along with determinants of lymphocyte trafficking are likely involved in the multifaceted cascade of immunological events that culminates in clinical aGVHD. T regulatory and Th17 cells are T cell subsets distinct from Th1 cells that are likely involved with aGVHD. Regulatory T cells (Tregs) have been implicated in the prevention of aGVHD in both mouse and man, while Th17 cells may modulate early inflammatory responses associated with aGVHD, especially those involving the skin and the lungs. Interestingly, these two lymphocyte subsets appear to be reciprocally regulated in part through retinoic acid, through cytokines such as IL-6, and via interactions with dendritic cells. Another area under tight regulation appears to be the homing of lymphocytes to lymph nodes, skin, and gut. Adhesion molecules including chemokine receptors, selectins, and integrins may identify specific T cell subsets with unique migratory functional properties during HSCT. Controlling the migration patterns of Th17 cells and Tregs represents a potential therapeutic target. A major goal of HSCT research will be to develop approaches to pharmacologically manipulate T cell subsets in vivo or to select, expand, and infuse T cell subsets that will maximize the targeted graft vs. tumor effect while minimizing the potentially fatal side effects of aGVHD. A better understanding of Tregs and their tissue specificity should lead to improvement in the success of HSCT.


[Reduction in the consumption of benzodiazepines due to a letter to chronic users from their own general practitioner].
To investigate the effectiveness of a minimal intervention in reducing the volume of prescription of benzodiazepines at the regional level. Prospective. Patients on compulsory health insurance who had received 180 or more daily doses of benzodiazepines in the course of one year received a letter from their general practitioner (GP) with information about the risks of chronic use, the advice to reduce or stop use, and an invitation to make an appointment to discuss the problem. This intervention took place in 19 GP practices in East Groningen. A reminder was sent six months later to the patients who had not responded in 9 randomly selected GP-practices. Thirty-seven practices in East Groningen and 91 practices in Northwest Groningen served as controls. Outcome measures were: (a) the percentage of patients who stopped, and (b) the change in average benzodiazepine consumption. During the period 6-12 months after the first letter in the intervention group (n = 1343), 11.3% of the patients (95% CI: 9.6-13.1) received no prescription whatsoever for benzodiazepines compared to 5.4% (4.6-6.3) and 4.9% (4.2-5.5) in East Groningen (n = 2932) and in Northwest Groningen (n = 4562), respectively. The average decrease in prescription volume was 13% (-9.9 to -15.1) in the intervention group compared to 3% (-0.1 to -4.1) and 3% (-1.5 to -4) in the control groups. The reminder sent half a year later had no additional effect. The informative letter from the GP to chronic users of benzodiazepines with the advice to stop or reduce this use was effective.


Advances in the determination of inorganic anions by ion chromatography.
The time period covered for this review includes articles published from 1997 to 1999, with the addition of a few classic references. The purpose of the review is to include the most relevant works from each topic area of the determination of inorganic anions by ion chromatography, including new sample pretreatments, new separation methods, new detection systems and the latest applications in the field of environmental, water, foods, etc. samples. Experimental conditions such as stationary phase, eluent, detection mode, as well as matrix are summarized in a table.


[Enzyme inhibitors from actinomycetes. II. Acid protease inhibitors].
In the course of a screening program directed toward the isolation and evaluation of inhibitors of carboxyl (acid) proteinases, we have found 0.39% of the investigated strains of microorganisms as producers of pepsin inhibitors. In more than 1543 strains which were isolated from various geographical areas and deposited with the IMET Culture Collection, Jena-GDR, and which belong to different genera of actinomycetes such as Streptomyces, Actinomadura, Actinoplanes, Micromonospora and non-identified strains, we have found 6 Streptomyces strains only. Surprisingly, the screening efficiency of the used casein agar spot test and the fibrin-blue-technique was very low. As compared with the inhibitor activity of pepstatin the yield of the fermentation broth of 3 wild type strains ranged from 6 to 27 micrograms/ml. A significant influence of the composition of culture fluids on the inhibitor yield was observed. Supplements by crude glutamine or mixtures of amino acids in culture fluids increased the yield from 3 to 15 times as compared with that of fermentation broth without supplements. After NMG-mutagenic treatment and use of familiar selection procedures 3.1% inhibitor-blocked mutants were detected among 480 strains selected.


Adjuvant and neoadjuvant radiotherapy and concurrent radiochemotherapy for rectal cancer.
Radical surgery with negative margins remains the most important prognostic factor in the treatment of rectal cancer. Combined modality treatment is the recommended standard adjuvant therapy for patients with locally advanced rectal cancer in the USA and in Germany. During the last decade substantial progress has been made in treatment modalities: surgical management currently includes a broad spectrum of operative procedures ranging from radical operations to innovative sphincter-preserving techniques. Specialized groups have reported excellent local control rates with total mesorectal excision (TME) alone. New and improved radiation techniques (conformal radiotherapy, intraoperative radiotherapy) and innovative schedules (protracted intravenous infusion, chronomodulated infusion) and combinations (oxaliplatin, irinotecan) of chemotherapy may have the potential to further increase the therapeutic benefit of adjuvant treatment. Moreover, the basic issue of timing of radio-(chemo-)therapy - preoperative versus postoperative - within a multimodality regimen is currently being addressed in prospective trials. Evidently, the current monolithic approaches, established by studies conducted more than a decade ago, to apply either the same schedule of postoperative radiochemotherapy to all patients with stage II/III rectal cancer or to give preoperative intensive short-course radiation according to the Swedish concept for all patients with resectable rectal cancer irrespective of tumor stage and treatment goal (e.g. sphincter preservation), need to be questioned. This review will discuss different irradiation settings in more recent and ongoing studies of perioperative radiotherapy for rectal cancer and will focus on the issue which patient should receive radiotherapy at all, and if so, how and when?


The impact of a character posture model on the communication of affect in an immersive virtual environment.
This paper presents the quantitative and qualitative findings from an experiment designed to evaluate a developing model of affective postures for full-body virtual characters in immersive virtual environments (IVEs). Forty-nine participants were each requested to explore a virtual environment by asking two virtual characters for instructions. The participants used a CAVE-like system to explore the environment. Participant responses and their impression of the virtual characters were evaluated through a wide variety of both quantitative and qualitative methods. Combining a controlled experimental approach with various data-collection methods provided a number of advantages such as providing a reason to the quantitative results. The quantitative results indicate that posture plays an important role in the communication of affect by virtual characters. The qualitative findings indicated that participants attribute a variety of psychological states to the behavioral cues displayed by virtual characters. In addition, participants tended to interpret the social context portrayed by the virtual characters in a holistic manner. This suggests that one aspect of the virtual scene colors the perception of the whole social context portrayed by the virtual characters. We conclude by discussing the importance of designing holistically congruent virtual characters especially in immersive settings.


Ammonification in Bacillus subtilis utilizing dissimilatory nitrite reductase is dependent on resDE.
During anaerobic nitrate respiration Bacillus subtilis reduces nitrate via nitrite to ammonia. No denitrification products were observed. B. subtilis wild-type cells and a nitrate reductase mutant grew anaerobically with nitrite as an electron acceptor. Oxygen-sensitive dissimilatory nitrite reductase activity was demonstrated in cell extracts prepared from both strains with benzyl viologen as an electron donor and nitrite as an electron acceptor. The anaerobic expression of the discovered nitrite reductase activity was dependent on the regulatory system encoded by resDE. Mutation of the gene encoding the regulatory Fnr had no negative effect on dissimilatory nitrite reductase formation.


Users' guides to the medical literature: XXIII. Qualitative research in health care A. Are the results of the study valid? Evidence-Based Medicine Working Group.
Quantitative research is designed to test well-specified hypotheses, determine whether an intervention did more harm than good, and find out how much a risk factor predisposes persons to disease. Equally important, qualitative research offers insight into emotional and experiential phenomena in health care to determine what, how, and why. There are 4 essential aspects of qualitative analysis. First, the participant selection must be well reasoned and their inclusion must be relevant to the research question. Second, the data collection methods must be appropriate for the research objectives and setting. Third, the data collection process, which includes field observation, interviews, and document analysis, must be comprehensive enough to support rich and robust descriptions of the observed events. Fourth, the data must be appropriately analyzed and the findings adequately corroborated by using multiple sources of information, more than 1 investigator to collect and analyze the raw data, member checking to establish whether the participants' viewpoints were adequately interpreted, or by comparison with existing social science theories. Qualitative studies offer an alternative when insight into the research is not well established or when conventional theories seem inadequate. JAMA. 2000;284:357-362


Osteogenic activity of OP-1 bone morphogenetic protein (BMP-7) in a human fibular defect.
We performed a prospective, randomised double-blind study in 24 patients undergoing high tibial osteotomy to evaluate the effectiveness of human recombinant osteogenic protein (OP-1) on a collagen type-I carrier in a critically-sized fibular defect. The study had two phases, each evaluated by clinical, radiological and DEXA methods during the first postoperative year. The first concerned the validation of the model of the fibular defect, using positive (demineralised bone) and negative (untreated) controls. The second phase concerned the osteogenic potential of OP-1 on collagen type-I v collagen type-I alone. The results of the first phase established the critically-sized nature of the defect. In the untreated group no bony changes were observed while, in the demineralised bone group, formation of new bone was visible from six weeks onwards. The results of the second phase showed no significant formation of new bone in the presence of collagen alone, while in the OP-1 group, all patients except one showed formation of new bone from six weeks onwards. This proved the osteogenic activity of OP-1 in a validated critically-sized human defect.


Mutations in isocitrate dehydrogenase 1 and 2 occur frequently in intrahepatic cholangiocarcinomas and share hypermethylation targets with glioblastomas.
Mutations in the genes encoding isocitrate dehydrogenase, IDH1 and IDH2, have been reported in gliomas, myeloid leukemias, chondrosarcomas and thyroid cancer. We discovered IDH1 and IDH2 mutations in 34 of 326 (10%) intrahepatic cholangiocarcinomas. Tumor with mutations in IDH1 or IDH2 had lower 5-hydroxymethylcytosine and higher 5-methylcytosine levels, as well as increased dimethylation of histone H3 lysine 79 (H3K79). Mutations in IDH1 or IDH2 were associated with longer overall survival (P=0.028) and were independently associated with a longer time to tumor recurrence after intrahepatic cholangiocarcinoma resection in multivariate analysis (P=0.021). IDH1 and IDH2 mutations were significantly associated with increased levels of p53 in intrahepatic cholangiocarcinomas, but no mutations in the p53 gene were found, suggesting that mutations in IDH1 and IDH2 may cause a stress that leads to p53 activation. We identified 2309 genes that were significantly hypermethylated in 19 cholangiocarcinomas with mutations in IDH1 or IDH2, compared with cholangiocarcinomas without these mutations. Hypermethylated CpG sites were significantly enriched in CpG shores and upstream of transcription start sites, suggesting a global regulation of transcriptional potential. Half of the hypermethylated genes overlapped with DNA hypermethylation in IDH1-mutant gliobastomas, suggesting the existence of a common set of genes whose expression may be affected by mutations in IDH1 or IDH2 in different types of tumors.


Ethical Inclusion of Health Care Workers in Covid-19 Research.
Employees are often considered a vulnerable research population due to concerns about consent and confidentiality, but there is insufficient guidance regarding their ethical inclusion in research. In the context of Covid-19, frontline health care workers comprise a particularly relevant research population in light of their risks of viral exposure and psychological strain, among other factors. They may therefore be targeted for research conducted at their place of employment and benefit from participating in such research. Beyond Covid-19, there are other circumstances in which health care workers may be considered for inclusion in research conducted by or with the involvement of their colleagues and employers. As investigators, sponsors, institutional review boards, and others assess the ethical permissibility of these scenarios, as well as relevant protections, we recommend systematic consideration of social and scientific value, validity, fairness, risks and benefits, voluntary consent, respect, and independent review. There is often good reason to specifically target health care workers for inclusion in Covid-19 research (beyond convenience), and they should not be excluded from research offering the prospect of direct benefit. However, additional safeguards may be necessary in employer-based research to avoid scientific bias, promote voluntariness, and solicit stakeholder input. Research personnel should be permitted to enroll in their own Covid-19 studies only when participation offers them the prospect of unique benefits.


Global gene expression analysis of murine limb development.
Detailed information about stage-specific changes in gene expression is crucial for understanding the gene regulatory networks underlying development and the various signal transduction pathways contributing to morphogenesis. Here we describe the global gene expression dynamics during early murine limb development, when cartilage, tendons, muscle, joints, vasculature and nerves are specified and the musculoskeletal system of limbs is established. We used whole-genome microarrays to identify genes with differential expression at 5 stages of limb development (E9.5 to 13.5), during fore- and hind-limb patterning. We found that the onset of limb formation is characterized by an up-regulation of transcription factors, which is followed by a massive activation of genes during E10.5 and E11.5 which levels off at later time points. Among the 3520 genes identified as significantly up-regulated in the limb, we find ~30% to be novel, dramatically expanding the repertoire of candidate genes likely to function in the limb. Hierarchical and stage-specific clustering identified expression profiles that are likely to correlate with functional programs during limb development and further characterization of these transcripts will provide new insights into specific tissue patterning processes. Here, we provide for the first time a comprehensive analysis of developmentally regulated genes during murine limb development, and provide some novel insights into the expression dynamics governing limb morphogenesis.


p38 MAPK activation controls the TLR3-mediated up-regulation of cytotoxicity and cytokine production in human NK cells.
Natural killer (NK) cells are a component of the innate immunity against viral infections through their rapid cytotoxic activity and cytokine production. Although the synthetic double-stranded (ds) RNA polyinosinic-polycytidylic acid (poly I:C), a mimic of a common product of viral infections, is known to rapidly up-regulate their in vivo functions, NK cell ability to directly respond to dsRNA is still mostly unknown. Our results show that treatment with poly I:C significantly up-regulates both natural and CD16-mediated cytotoxicity of highly purified human NK cells. Poly I:C also induces the novel capability of producing CXCL10 chemokine in human NK cells and synergistically enhances interferon-gamma (IFN-gamma) production induced by either adaptive or innate cytokines. In accordance with the expression of Toll-like receptor-3 (TLR3) and of TRIF/TICAM-1 adaptor, poly I:C stimulation induces the activation of interferon regulatory factor-3 (IRF-3) transcription factor and of p38 mitogen-activated protein kinase (MAPK) in human NK cells. Finally, we demonstrate that p38 MAPK activity is required for the dsRNA-dependent enhancement of cytotoxicity and CXCL10 production. The occurrence of dsRNA-induced signaling and functional events closely correlates with the TLR3 mRNAprofile in different NK cell populations. Taken together, these data identify p38 as a central component of NK cell ability to directly respond to dsRNA pathogen-associated molecular pattern (PAMP).


Correlation between reference point indentation and mechanical properties of 3D-printed polymers.
Reference point indentation (RPI) is a novel experimental technique designed to evaluate bone quality. This study utilizes two RPI instruments, BioDent and Osteoprobe, to investigate the mechanical responses of several 3D-printed polymers. We correlated the mechanical properties from a tensile test with the RPI parameters obtained from the BioDent and OsteoProbe. In addition, we tested the same polymers five years later (Age 5). The results show that for Age 0 polymers, the elastic modulus is highly correlated with average unloading slope (r = 0.87), first unloading slope (r = 0.85), bone material strength index (BMSi) (r = 0.85), average loading slope (r = 0.82), first indentation distance (r = 0.79), and total indentation distance (r = 0.76). The ultimate stress correlates significantly with first unloading slope (r = 0.85), average unloading slope (r = 0.83), BMSi (r = 0.81), first indentation distance (r = 0.73), average loading slope (r = 0.71), and total indentation distance (r = 0.70). The elongation has no significant correlation with the RPI parameters except with the average creep indentation distance (r = 0.60). For Age 5 polymers, correlations between mechanical properties and RPI parameters are low. This study illustrates the potential of RPI to assess the mechanical properties of polymers nondestructively with simple sample requirements. Furthermore, for the first time, 3D-printed polymers and aged polymers are investigated with RPI.


Communication strategies and media discourses in the age of COVID-19: an urgent need for action.
Identified in December 2019 in China, the coronavirus 2019 (COVID-19) has been declared a Public Health Emergency of International Concern (PHEIC). Pandemics share features that increase fear. While some fear can stimulate preventive health behaviors, extreme fear can lead to adverse psychological and behavioral response. The media play a major role shaping these responses. When dealing with a PHEIC, the authorities' communication strategies are embedded in a multilevel governance and a highly hierarchal system, which adds another layer of complexity. Carrying out more 'real-world research' is crucial to generate evidence relating to the psychosocial and behavioral aspects involved during the COVID-19 pandemic and how it is shaped by authorities and media discourses. Interdisciplinary research and international collaborations could contribute to improve our understanding and management of risk information. Emerging from a socio-ecological perspective, future research must integrate multilevel analytical elements, to ensure triangulation of evidence and co-constructing robust recommendations. A mixed-method approach should be privileged to address these issues. At the micro-level, a population-based survey could be conducted in various jurisdictions to assess and compare psychosocial issues according to sociocultural groups. Then, a quantitative/qualitative discourse analysis of the media could be performed. Finally, a network analysis could allow assessing how official information flows and circulates across levels of governance. The COVID-19 represents an opportunity to evaluate the impacts of information/communication strategy and misinformation on various cultural and socioeconomic groups, providing important lessons that could be applied to future health emergencies and disasters.


Identification of a novel glucocorticoid receptor mutation in budesonide-resistant human bronchial epithelial cells.
We developed a molecular genetic model to investigate glucocorticoid receptor (GR) signaling in human bronchial epithelial cells in response to the therapeutic steroid budesonide. Based on a genetic selection scheme using the human Chago K1 cell line and integrated copies of a glucocorticoid-responsive herpes simplex virus thymidine kinase gene and a green fluorescent protein gene, we isolated five Chago K1 variants that grew in media containing budesonide and ganciclovir. Three spontaneous budesonide-resistant subclones were found to express low levels of GR, whereas two mutants isolated from ethylmethane sulfonate-treated cultures contained normal levels of GR protein. Analysis of the GR coding sequence in the budesonide-resistant subclone Ch-BdE5 identified a novel Val to Met mutation at amino acid position 575 (GRV575M) which caused an 80% decrease in transcriptional regulatory functions with only a minimal effect on ligand binding activity. Homology modeling of the GR structure in this region of the hormone binding domain and molecular dynamic simulations suggested that the GRV575M mutation would have a decreased affinity for the LXXLL motif of p160 coactivators. To test this prediction, we performed transactivation and glutathione-S-transferase pull-down assays using the p160 coactivator glucocorticoid interacting protein 1 (GRIP1)/transcriptional intermediary factor 2 and found that GRV575M transcriptional activity was not enhanced by GRIP1 in transfected cells nor was it able to bind GRIP1 in vitro. Identification of the novel GRV575M variant in human bronchial epithelial cells using a molecular genetic selection scheme suggests that functional assays performed in relevant cell types could identify subtle defects in GR signaling that contribute to reduced steroid sensitivities in vivo.


Artificial metal with effective plasma frequency in near-infrared region.
We have proposed and demonstrated an artificial medium consisting of arrays of circular metal rods embedded in a dielectric host, which holds a real metal behavior but the extracted effective plasma frequency is in near-infrared region. The electromagnetic responses of such medium and the retrieved effective material parameters have been particularly shown. In addition, an analytic model about effective plasma frequency is constructed by uniquely considering the skin effect and introducing the parameter-skin depth, whose predicting results are in well accordance with the FDTD simulation. This artificial material may open possibilities for many metal-based applications in near-infrared regime.


The Case for Extragenital Screening of Chlamydia trachomatis and Neisseria gonorrhoeae in the College Health Setting.
Although the Centers for Disease Control and Prevention does not recommend routine oropharyngeal and anorectal screening for Chlamydia trachomatis and Neisseria gonorrhoeae in the general population, they do recommend it for men who have sex with men. However, risk-based extragenital screening of men may not have been adopted at all college health centers, and existing research has not focused on the college population. We examined health records of men at a college health center in a large urban university over 6 years to evaluate effectiveness of C. trachomatis and N. gonorrhoeae screening. We also evaluated the proportion of C. trachomatis and N. gonorrhoeae infections that would have been missed if risk-based extragenital screening were not performed. Decisions to screen at extragenital sites were based on patient-reported risk behavior. For 4093 male college students screened, 7.6% of the screening visits used extragenital screening in response to self-reported risk behaviors. The case positivity rate for C. trachomatis was 3.1% with urogenital-only screening and 3.7% with risk-prompted extragenital screening. The case positivity rate for N. gonorrhoeae was 0.7% with urogenital-only screening and 3.3% with risk-prompted extragenital screening. If the college health center had relied solely on urogenital screening rather than risk-based extragenital screening, 26.4% of C. trachomatis infections and 63.2% of N. gonorrhoeae infections would have been missed. One out of four C. trachomatis infections and 2 of 3 N. gonorrhoeae infections would have been missed without extragenital screening in this analysis of college men. This study reinforces Centers for Disease Control and Prevention recommendations for risk-based extragenital screening and is the first report to focus on college men. Because guidelines exist only for men, future studies should focus on extragenital screening in college women to build evidence for another group of patients that may benefit from this practice, given the high risk in young adults.


Improving the operating budget process at Abbott Northwestern Hospital.
To set an example for the organization, the senior management team at Abbott Northwestern Hospital used quality tools and techniques to improve a highly visible, problematic system--the operating budget process. Team members narrowed the root cause to a faulty, nonintegrated assumption process and identified additional problems, such as inadequate communication, unclear accountabilities, and lengthy timetables. Then, the team streamlined the budget process from a 6-month to a 10-week cycle, which aligned the process with one of the organization's shared quality goals--reducing cycle time. Cycle time was reduced by 60 percent, from an average of 6 months to 10 weeks. Budget accuracy, which is measured by how close initial budget submissions come to targeted net income projections, improved by $12 million or 500 percent. The amount of rework, which is measured by the number of budget revisions made prior to Board approval, dropped from eight revisions to four. Overall satisfaction and familiarity with the budget process increased by 40 percent among managers and supervisors.


GDF15 is a major determinant of ketogenic diet-induced weight loss.
A ketogenic diet (KD) has been promoted as an obesity management diet, yet its underlying mechanism remains elusive. Here we show that KD reduces energy intake and body weight in humans, pigs, and mice, accompanied by elevated circulating growth differentiation factor 15 (GDF15). In GDF15- or its receptor GFRAL-deficient mice, these effects of KD disappeared, demonstrating an essential role of GDF15-GFRAL signaling in KD-mediated weight loss. Gdf15 mRNA level increases in hepatocytes upon KD feeding, and knockdown of Gdf15 by AAV8 abrogated the obesity management effect of KD in mice, corroborating a hepatic origin of GDF15 production. We show that KD activates hepatic PPARγ, which directly binds to the regulatory region of Gdf15, increasing its transcription and production. Hepatic Pparγ-knockout mice show low levels of plasma GDF15 and significantly diminished obesity management effects of KD, which could be restored by either hepatic Gdf15 overexpression or recombinant GDF15 administration. Collectively, our study reveals a previously unexplored GDF15-dependent mechanism underlying KD-mediated obesity management.


The folate precursor para-aminobenzoic acid elicits induced resistance against Cucumber mosaic virus and Xanthomonas axonopodis.
The use of vitamins including vitamin B1, B2 and K3 for the induction of systemic acquired resistance (SAR) to protect crops against plant pathogens has been evaluated previously. The use of vitamins is beneficial because it is cost effective and safe for the environment. The use of folate precursors, including ortho-aminobenzoic acid, to induce SAR against a soft-rot pathogen in tobacco has been reported previously. In the present study, para-aminobenzoic acid (PABA, also referred to as vitamin Bx) was selected owing to its effect on the induction of SAR against Xanthomonas axonopodis pv. vesicatoria in pepper plants through greenhouse screening. Dipping of pepper seedlings in a 1 mm PABA solution in field trials induced SAR against artificially infiltrated X. axonopodis pv. vesicatoria and naturally occurring cucumber mosaic virus. Expression of the Capsicum annuum pathogenesis-related 4 gene was primed in response to pathogen infection as assessed by quantitative real-time PCR. The accumulation of cucumber mosaic virus RNA was reduced in PABA-treated pepper plants at 40 and 105 d post-treatment. Unexpectedly, fruit yield was increased in PABA-treated plants, indicating that PABA-mediated SAR successfully protected pepper plants from infection by bacterial and viral pathogens without significant fitness allocation costs. The present study is the first to demonstrate the effective elicitation of SAR by a folate precursor under field conditions.


Influenza A/H1N1v in pregnancy: an investigation of the characteristics and management of affected women and the relationship to pregnancy outcomes for mother and infant.
In April 2009 a novel influenza A virus (AH1N1v) of swine origin (swine flu) emerged, spreading rapidly and achieving pandemic status in June 2009. Pregnant women were identified as being at high risk of severe influenza-related complications and as a priority group for vaccination against AH1N1v. Limited information was available about the maternal and fetal risks of AH1N1v infection or of antiviral drug or AH1N1v vaccine use in pregnancy. To assess rates of and risk factors for adverse outcomes following AH1N1v infection in pregnancy and to assess the adverse effects of the antiviral drugs and vaccines used in prevention and management. Prospective national cohort studies were conducted to identify pregnant women who were (1) suspected to be infected with AH1N1v or being treated with antiviral medication in primary care; (2) vaccinated against AH1N1v; and (3) admitted to hospital with confirmed AH1N1v. Characteristics of women with influenza-like illness (ILI) in primary care were compared with those of women without symptoms accepting or declining immunisation. Characteristics of women admitted to hospital with confirmed AH1N1v infection in pregnancy were compared with a historical cohort of over 1200 women giving birth in the UK who were uninfected with AH1N1v. Outcomes examined in hospitalised women included maternal death, admission to an intensive care unit, perinatal mortality and preterm birth. Risk factors for hospital and intensive care unit admission were examined in a full regression model. The weekly incidence of ILI among pregnant women averaged 51/100,000 over the study period. Antiviral drugs were offered to 4.8% [95% confidence interval (CI) 4.0% to 5.9%] and vaccination to 64.8% (95% CI 64.7% to 68.9%) of registered pregnant women. Ninety pregnant women with ILI presenting in primary care were reported to the research team, 55 of whom were prescribed antiviral drugs and in 42 (76%) cases this was within 2 days of symptom onset. After comparison with 1329 uninfected pregnant women offered vaccination, pre-existing asthma was the only maternal factor identified as increasing risk of ILI presentation [adjusted odds ratio (OR) 2.0, 95% CI 1.0 to 3.9]. Maternal obesity and smoking during pregnancy were also associated with hospital admission with AH1N1v infection. Overall, 241 pregnant women were admitted to hospital with laboratory-confirmed AH1N1v infection. Eighty-three per cent of these women were treated with antiviral agents, but only 6% received antiviral treatment before hospital admission. Treatment within 2 days of symptom onset was associated with an 84% reduction in the odds of admission to an intensive therapy unit (OR 0.16, 95% CI 0.08 to 0.34). Women admitted to hospital with AH1N1v infection were more likely to deliver preterm; a three times increased risk was suggested compared with an uninfected population cohort (OR 3.1, 95% CI 2.1 to 4.5). Earlier treatment with antiviral agents is associated with improved outcomes for pregnant women and further actions are needed in future pandemics to ensure that antiviral agents and vaccines are provided promptly to pregnant women, particularly in the primary care setting. Further research is needed on longer-term outcomes for infants exposed to AH1N1v influenza, antiviral drugs or vaccines during pregnancy.


The relationship between maternal smoking during pregnancy and parental-reported experience of dental caries in Indigenous Australian children.
This study aimed to test the association between maternal smoking during pregnancy and parental-reported experience of dental caries in Indigenous Australian children. Data were from the Longitudinal Study of Indigenous Children (LSIC); a population-based cohort study in Australia. Participants were 1,687 Indigenous Australian children aged 5 or less. Biological, social and behavioural variables were tested using log-linear modelling with binomial regression to determine the association with parental-reported experience of dental caries. Markov Chain Monte Carlo methods were used for multiple imputation of missing data. Overall 25.8% of Indigenous Australian children had dental caries as reported by a carer. In the multivariable model, increased prevalence of parental-reported caries was significantly associated with low maternal education levels (RR=1.60, 95%CI 1.17,2.20) and high sugar consumption (RR= 1.60, 95%CI 1.26,2.02). In the group of children whose mothers smoked tobacco during pregnancy, the association with parent-reported dental caries approached the threshold of significance, but was not significantly associated with caries status in children (RR=1.19, 95%CI 0.99,1.43). After multiple imputation, the most significant association was evident in children of the least educated mothers (RR=1.57, 95%CI 1.25,1.95), breastfeeding more than 12 months (RR=1.26, 95%CI 1.01,1.56), sweet intake more than 30% (RR=1.42, 95%CI 1.15,1.74) and 20-30% (RR=1.29 95%CI 1.04,1.59) and residing in outer regional (RR=1.56, 95%CI 1.19,2.05) or inner regional locations (RR=1.50, 95%CI 1.19,1.88). Mothers' tobacco smoking status showed a weak association with parent-reported dental decay (RR=1.42, 95%CI 1.20,1.68). This study suggests there is a weak association between maternal smoking during pregnancy and prevalence of parentally-reported dental caries in Indigenous Australian children.


In vivo Cross-Linking MS of the Complement System MAC Assembled on Live Gram-Positive Bacteria.
Protein-protein interactions are central in many biological processes, but they are challenging to characterize, especially in complex samples. Protein cross-linking combined with mass spectrometry (MS) and computational modeling is gaining increased recognition as a viable tool in protein interaction studies. Here, we provide insights into the structure of the multicomponent human complement system membrane attack complex (MAC) using in vivo cross-linking MS combined with computational macromolecular modeling. We developed an affinity procedure followed by chemical cross-linking on human blood plasma using live Streptococcus pyogenes to enrich for native MAC associated with the bacterial surface. In this highly complex sample, we identified over 100 cross-linked lysine-lysine pairs between different MAC components that enabled us to present a quaternary model of the assembled MAC in its native environment. Demonstrating the validity of our approach, this MAC model is supported by existing X-ray crystallographic and electron cryo-microscopic models. This approach allows the study of protein-protein interactions in native environment mimicking their natural milieu. Its high potential in assisting and refining data interpretation in electron cryo-tomographic experiments will be discussed.


The Neurosurgical Workforce of the Philippines.
No study has comprehensively examined the delivery of neurosurgical care in the Philippines, a lower-middle-income country in Southeast Asia with a total population of 109 million. We aimed to quantify the workforce, map the distribution, and characterize the clinical practice of neurosurgeons across the 17 regions and 81 provinces of the Philippines. An online survey was sent to all fellows of the Academy of Filipino Neurosurgeons and all graduates of neurosurgical training programs in the country. Neurosurgeons who have been in active clinical practice for at least 1 year were eligible to participate. A database of Filipino neurosurgeons was generated through personal communications, correspondence with neurosurgery departments, and accessing publicly available information. The top neurosurgical procedures were identified to estimate the overall volume of neurosurgical disease. There are 174 neurosurgeons practicing in the Philippines or approximately one neurosurgeon for every 600 000 people. In 9 provinces, neurosurgeons were only available part-time, and 35 provinces had no neurosurgeons at all, equivalent to an underserved population of 24 million people. Among 99 survey respondents, the median numbers of neurosurgical consults and operations every month were 30 (IQR:35) and 8 (IQR:8), respectively. The top neurosurgical procedures were burr holes/craniotomy for traumatic brain injury, craniotomy for stroke, and biopsy/resection of brain tumors. There are an estimated 93 498 cases requiring essential neurosurgery every year. Although positive trends have been observed in the number, distribution, and composition of neurosurgeons in the Philippines, there remains a large workforce deficit that needs to be addressed to provide timely, quality, and affordable neurosurgical care to the entire population.


Characterization of fibroblasts recruited from bone marrow-derived precursor in neonatal bronchopulmonary dysplasia mice.
We sought to determine whether the extrapulmonary origin of fibroblasts derived from bone marrow (BM) progenitor cells is essential to lung fibrosis in bronchopulmonary dysplasia (BPD). Neonate mice were durably engrafted with BM isolated from transgenic reporter mice that expressed green fluorescent protein (GFP). Such chimera mice were subjected to 60% O(2) exposure for 14 days. A large number of fibroblast-specific protein-1 (FSP1) and GFP-positive fibroblasts were identified in active fibrotic lesions. More surprisingly, however, FSP1(+) fibroblasts also arose in considerable numbers from BM-derived alveolar type II cells (AT2) through epithelial-mesenchymal transition (EMT) during lung fibrogenesis. Cultured lung fibroblasts could express the CXC chemokine receptor (CXCR4) and responded chemotactically to their cognate ligand, chemokine (C-X-C motif) ligand 12 (CXCL12), which were elevated in the serum of BPD mice. These data suggest that lung fibroblasts in BPD fibrosis could variably arise from BM progenitor cells. This finding, which suggests the pathophysiological process of fibrosis, could contribute to a therapy for BPD that is characterized by extensive interstitial fibrosis.


Animal Models of Type 2 Diabetes Complications: A Review.
Diabetes mellitus is a multifactorial metabolic disease, of which type 2 diabetes (T2D) is one of the most common. The complications of diabetes are far more harmful than diabetes itself. Type 2 diabetes complications include diabetic nephropathy (DN), diabetic heart disease, diabetic foot ulcers (DFU), diabetic peripheral neuropathy (DPN), and diabetic retinopathy (DR) et al. Many animal models have been developed to study the pathogenesis of T2D and discover an effective strategy to treat its consequences. In this sense, it is crucial to choose the right animal model for the corresponding diabetic complication. This paper summarizes and classifies the animal modeling approaches to T2D complications and provides a comprehensive review of their advantages and disadvantages. It is hopeful that this paper will provide theoretical support for animal trials of diabetic complications.


A new blind loach species, Triplophysa huanjiangensis (Teleostei: Balitoridae), from Guangxi, China.
A new blind loach of the genus Triplophysa was collected in 2007 from a cave in Leyi village, located near Chuanshan Town, Huangjiang County in the Guangxi Zhuang Autonomous Region of southern China. The new species, Triplophysa huangjiangensis sp. nov, can be distinguished from its congeners by eyes and scales absent; caudal-fin forked; adipose keel present; lips with furrows; distal margin of dorsal-fin truncate, dorsal fin origin much closer to caudal-fin base than snout tip; 8-9 branched dorsal-fin rays; 6-7 branched pelvic-fin rays; 6 branched anal-fin rays; 10-11 branched pectoral-fin rays; 13-14 branched caudal-fin rays; pectoral-fin length 52.3%-70.7% the distance between pectoral-fin origin to pelvic-fin origin; dorsal-fin origin posterior to vertical line of pelvic-fin origin; outer rostral barbel longer than other two pairs of barbels, with 47.0%-73.8% of lateral head length; anterior nostril with elongate barbel-like tip; and posterior chamber of gas-bladder developed, reaching pelvic-fin origin.


Microperimetry and fundus autofluorescence in diabetic macular edema: subthreshold micropulse diode laser versus modified early treatment diabetic retinopathy study laser photocoagulation.
The purpose of this study was to evaluate and compare microperimetry and fundus autofluorescence (FAF) after subthreshold micropulse diode laser versus modified Early Treatment Diabetic Retinopathy Study photocoagulation for clinically significant diabetic macular edema. A prospective randomized clinical trial including 62 eyes (50 patients) with untreated, center-involving, clinically significant diabetic macular edema was performed. All patients underwent best-corrected visual acuity determination (logarithm of the minimum angle of resolution), slit-lamp biomicroscopy, FAF, optical coherence tomography, microperimetry (macular sensitivity), and fluorescein angiography before and after treatment. Best-corrected visual acuity, optical coherence tomography, microperimetry, and FAF were repeated at 1-, 3-, 6-, 9-, and 12-month follow-up examinations. Fluorescein angiography was performed at baseline and at 6 and 12 months. Before treatment, demographic and macular parameters were not different between the two treatment groups. At 12 months, best-corrected visual acuity remained stable in both groups (P = 0.41 and P = 0.82), mean central retinal thickness decreased in both groups (P = 0.0002 and P < 0.0001), and mean central 4 degrees and 12 degrees retinal sensitivity increased in the micropulse diode laser group (P = 0.02 and P = 0.0075) and decreased in the Early Treatment Diabetic Retinopathy Study group (P = 0.2 and P = 0.0026). There was no significant difference in either best-corrected visual acuity or central retinal thickness between the 2 treatment groups (P = 0.48 and P = 0.29), whereas there was a significant difference in 4 degrees and 12 degrees retinal sensitivity (P = 0.04 and P < 0.0001). Fundus autofluorescence never changed in the micropulse diode laser group even after retreatment. In the Early Treatment Diabetic Retinopathy Study group, FAF increased up to 9 months and decreased in 6 eyes (20%) at 12 months. Micropulse diode laser seems to be as effective as modified Early Treatment Diabetic Retinopathy Study laser photocoagulation in the treatment of clinically significant diabetic macular edema. Micropulse diode laser treatment does not determine any change on FAF showing (at least) nonclinically visible damage of the retinal pigment epithelium. Microperimetry data encourage the use of a new, less aggressive laser therapeutic approach in the treatment of clinically significant diabetic macular edema.


[Needle stick injuries in health care - frequency, causes und preventive strategies].
Healthcare workers (HCW) are at risk for infections with blood-borne pathogens - especially hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) - resulting from occupational blood-exposure trough injuries with sharp instruments and needle sticks. Results of a study on the epidemiology of needle stick injuries (NSI) among HCW in two German hospitals indicate that 500,000 NSI occur annually in Germany. Most of these injuries occur during disposal of used syringes and "recapping". Administration of the post-exposure prophylaxis is recommended for HCW who are occupationally exposed to HBV (vaccine/immunoglobulin) and HIV (antiretroviral drugs) i.e. the immediately reporting of blood exposure is very important. Comprehensive programmes to prevent NSI - e.g. avoiding of recapping, use of disposal containers, surgical gloves and in particular safety devices - minimize a high cost of NSI due to the administration of PEP, developing of chronic hepatitis, cirrhosis and liver cancer.


Homologous monocuspid valve patch in right ventricular outflow tract reconstruction.
In the surgical repair of tetralogy of Fallot or pulmonary atresia, pulmonary regurgitation may be detrimental in the postoperative period. We have used homograft monocuspid valve patch to prevent pulmonary insufficiency. From September 1996 to December 1998, twenty-five patients, 4 months to 8 years of age (median 10.1 months) had homograft monocuspid valve in the procedure of right ventricular outflow tract reconstruction. The function of the monocuspid valve was assessed by echocardiogram and graded as trivial to mild, mild to moderate, moderate, and severe. We evaluated the degree of pulmonary insufficiency before discharge, at 3-6 months, and at 12 months after the operation. There was one hospital death due to fulminate adeno viral pneumonia. On echocardiogram, 21 patients (88%, 21/24) had no significant pulmonary insufficiency. Only one patient (4.5%) showed a moderate degree of pulmonary insufficiency. At 3-6 months, seventeen of twenty-one (81%) patients had no significant pulmonary insufficiency. There were fourteen patients who had follow-up over 1 year, and no patients showed newly developed significant pulmonary insufficiency. We concluded that the homograft monocuspid valve patch for right ventricular outflow tract reconstruction has provided excellent early results for the prevention of pulmonary insufficiency. However these effects are limited in duration and further close follow-up should be needed.


Primary central nervous system lymphoma presenting as bilateral tumors of the internal auditory canal.
The increasing incidence of central nervous system (CNS) lymphoma in the general population, especially in young adults with AIDS, should alert the otolaryngologist that in future there will be an even greater incidence of this rare disease. It may be responsible for hearing loss accompanied by other neurological deficits. CNS lymphoma often has a rapidly progressive course. Early diagnosis and rapid therapy are crucial for a better prognosis. We report a case of primary CNS lymphoma involving both internal auditory canals that presented with sudden deafness and disequilibrium accompanied by facial and abducens nerve palsy.


HPLC-DAD-qTOF Compositional Analysis of the Phenolic Compounds Present in Crude Tomato Protein Extracts Derived from Food Processing.
The conversion of raw fruits and vegetables, including tomatoes into processed food products creates side streams of residues that can place a burden on the environment. However, these processed residues are still rich in bioactive compounds and in an effort to valorize these materials in tomato by-product streams, the main aim of this study is to extract proteins and identify the main phenolic compounds present in tomato pomace (TP), peel and skins (TPS) by HPLC-DAD-ESI-QTOF. Forty different phenolic compounds were identified in the different tomato extracts, encompassing different groups of phenolic compounds, including derivatives of simple phenolic acid derivatives, hydroxycinnamoylquinic acid, flavones, flavonones, flavonol, and dihydrochalcone. In the crude protein extract (TPE) derived from tomatoes, most of these compounds were still present, confirming that valuable phenolic compounds were not degraded during food processing of these co-product streams. Moreover, phenolic compounds present in the tomato protein crude extract could provide a valuable contribution to the required daily intake of phenolics that are usually supplied by consuming fresh vegetables and fruits.