Stereoselective uptake of atenolol into storage granules isolated from PC12 cells.
Atenolol has been shown to be stored and secreted from PC12 cells by calcium-dependent and stereoselective mechanisms. The present study was designed to determine if the cytoplasmic amine storage granule was the site for storage and release of atenolol and if the drug was, in fact, an enantiomeric selective substrate for the vesicular amine transport protein. Uptake of racemic [3H]atenolol and (-)-[3H]norepinephrine was studied in a vesicular preparation of storage granules isolated from PC12 cells. Uptake of both molecules was found to be ATP-dependent (80%) and to reach steady state in approximately 10 to 15 min. Uptake and storage was inhibited (95%) by either reserpine, an inhibitor of the vesicular amine carrier, or by nigericin which dissipates the proton gradient across the membrane. Uptake of neither compound was inhibited by desipramine, an inhibitor of uptake 1, or oligomycin, an inhibitor of mitochondrial adenosine triphosphatase. Furthermore, uptake of the (-)-enantiomer of atenolol was found to be approximately 5-fold greater than the (+)-enantiomer. Kinetic studies revealed a Km of 184 microM for (+/-)-atenolol and 79 microM for (-)-norepinephrine and Vmax values of 750 and 503 pmol/min/mg of protein for atenolol and norepinephrine, respectively. The interaction of the two compounds with the transport process was found to be kinetically competitive. In separate experiments, atenolol was also found to be transported stereoselectively into bovine adrenal chromaffin granule ghosts and to be ATP dependent (85%) and reserpine sensitive (44%).(ABSTRACT TRUNCATED AT 250 WORDS)


Pyridoxamine analogues scavenge lipid-derived gamma-ketoaldehydes and protect against H2O2-mediated cytotoxicity.
Isoketals and levuglandins are highly reactive gamma-ketoaldehydes formed by oxygenation of arachidonic acid in settings of oxidative injury and cyclooxygenase activation, respectively. These compounds rapidly adduct to proteins via lysyl residues, which can alter protein structure/function. We examined whether pyridoxamine, which has been shown to scavenge alpha-ketoaldehydes formed by carbohydrate or lipid peroxidation, could also effectively protect proteins from the more reactive gamma-ketoaldehydes. Pyridoxamine prevented adduction of ovalbumin and also prevented inhibition of RNase A and glutathione reductase activity by the synthetic gamma-ketoaldehyde, 15-E2-isoketal. We identified the major products of the reaction of pyridoxamine with the 15-E2-isoketal, including a stable lactam adduct. Two lipophilic analogues of pyridoxamine, salicylamine and 5'-O-pentylpyridoxamine, also formed lactam adducts when reacted with 15-E2-isoketal. When we oxidized arachidonic acid in the presence of pyridoxamine or its analogues, pyridoxamine-isoketal adducts were found in significantly greater abundance than the pyridoxamine-N-acyl adducts formed by alpha-ketoaldehyde scavenging. Therefore, pyridoxamine and its analogues appear to preferentially scavenge gamma-ketoaldehydes. Both pyridoxamine and its lipophilic analogues inhibited the formation of lysyl-levuglandin adducts in platelets activated ex vivo with arachidonic acid. The two lipophilic pyridoxamine analogues provided significant protection against H2O2-mediated cytotoxicity in HepG2 cells. These results demonstrate the utility of pyridoxamine and lipophilic pyridoxamine analogues to assess the potential contributions of isoketals and levuglandins in oxidant injury and inflammation and suggest their potential utility as pharmaceutical agents in these conditions.


Insights into suckling rabbit feeding behaviour: acceptability of different creep feed presentations and attractiveness for sensory feed additives.
In young rabbit, digestive disorders are frequently observed around weaning. Stimulating the onset of feed intake in the suckling rabbit might be a way to promote gut health. The aim of this study was to determine the rabbit's acceptability for different feed presentations and its preferences for flavours at an early stage of life. Two trials were conducted to evaluate the effects of physical form and flavouring on creep feed attractiveness. All the diets tested were provided in the nest from 3 to 17 days, and the daily intake per litter was recorded as of 8 days of age. In the first trial, five feed presentations were tested separately (n = 60 litters). Three dry presentations were chosen: commercial pellet (P), crumb from commercial pellet (cP) and crumb from beet pulp pellet (cBP). Hydrated feeds were also provided with either raw fodder beetroot (B) or a semi-solid feed in agar gel form produced with fodder beetroot juice and pulp (gB). In the second trial, double-choice tests were performed on four feed gels (n = 72 litters), leading to six comparison treatments. These agar gels were made of pellet mash without or with a sensory additive: one non-odorised control gel and three gels with 0.20% banana flavour, 0.06% red berry flavour and 0.10% vanilla flavour, respectively. In the first trial, kits ate more gB in fresh matter than other feed presentations (P < 0.001), with a total intake of 7.0 ± 1.8 g/rabbit from 8 to 17 days. In DM, the total consumption of pellets P (1.6 ± 0.4 g of DM/rabbit) was the highest together with the gB form (1.4 ± 0.4 g of DM/rabbit), whereas cBP was barely consumed (0.3 ± 0.1 g of DM/rabbit). Gel feed supplemented with vanilla was slightly more consumed than other flavoured and non-odorised gels (relative consumption of 57% when compared to control gel; P = 0.001). The gel feed intake was independent of the milk intake but was correlated with the litter weight at 3 days (r = 0.40, P < 0.001). In both trials, rabbit growth before and after weaning was not affected by the type of creep feed provided. Our results confirmed that providing creep feed promotes the solid intake of rabbits at early stages. Gel feed form motivated rabbits to eat and vanilla flavour supplementation increased the feed palatability. Those creep feed characteristics should be explored further for seeking effective stimulation of the onset of the feed intake in suckling rabbit.


Analysis of deposit depth and morphology in granular corneal dystrophy type 2 using fourier domain optical coherence tomography.
Granular corneal dystrophy type 2 (GCD2) causes the formation of corneal deposits having 3 different morphological types. We used Fourier domain optical coherence tomography to assess the depths of each type according to the morphology. A prospective study was performed in 54 eyes of 54 heterozygous patients with GCD2. Corneal deposits of 54 patients with GCD2 were classified into 3 morphological types: type 1, diffuse haze; type 2, granular shape (2 subgroups: type 2a, round granulated and type 2b, round spiculated); and type 3, linear shape (2 subgroups: type 3a, short side branched and type 3b, long side branched). Using Fourier domain optical coherence tomography, we measured the distances from the Bowman layer to the upper surface of the deposits (USBL), to the lower surface of the deposits (LSBL), and the thickness of the deposits (TD). The deposits formed along the flap interface were also examined among 19 patients who had LASIK. Types 1 and 2 deposits were always adjacent to the Bowman layer; thus the USBLs for each were 0.0 ± 0.0 μm, whereas that of type 3 deposits was 65.4 ± 48.0 μm (P < 0.0001). The LSBL and TD of linear deposits with long side branches (type 3) (313.3 ± 71.4 and 246.2 ± 71.9 μm) were greater than those of type 1 (47.7 ± 10.2 and 47.7 ± 10.2 μm) and type 2 (91.3 ± 39.5 and 91.3 ± 39.5 μm) (P < 0.0001). There were no differences in the measurements between the subgroups type 2a and type 2b or between types 3a and 3b. USBL of the laser in situ keratomileusis group was 54.5 ± 29.8 μm. The depths of corneal deposits in patients with GCD2 were associated with the morphology of the deposits. The linear deposits were located most deeply in the cornea, followed by granular deposits and diffuse haze moving anteriorly. Several deposits have distinct depths according to the morphological types.


Activation of HIV-1 with Nanoparticle-Packaged Small-Molecule Protein Phosphatase-1-Targeting Compound.
Complete eradication of HIV-1 infection is impeded by the existence of latent HIV-1 reservoirs in which the integrated HIV-1 provirus is transcriptionally inactive. Activation of HIV-1 transcription requires the viral Tat protein and host cell factors, including protein phosphatase-1 (PP1). We previously developed a library of small compounds that targeted PP1 and identified a compound, SMAPP1, which induced HIV-1 transcription. However, this compound has a limited bioavailability in vivo and may not be able to reach HIV-1-infected cells and induce HIV-1 transcription in patients. We packaged SMAPP1 in polymeric polyethylene glycol polymethyl methacrylate nanoparticles and analyzed its release and the effect on HIV-1 transcription in a cell culture. SMAPP1 was efficiently packaged in the nanoparticles and released during a 120-hr period. Treatment of the HIV-1-infected cells with the SMAPP1-loaded nanoparticles induced HIV-1 transcription. Thus, nanoparticles loaded with HIV-1-targeting compounds might be useful for future anti-HIV-1 therapeutics.


Postinduction dexamethasone and individualized dosing of Escherichia Coli L-asparaginase each improve outcome of children and adolescents with newly diagnosed acute lymphoblastic leukemia: results from a randomized study--Dana-Farber Cancer Institute ALL Consortium Protocol 00-01.
We assessed the toxicity and efficacy of dexamethasone and a novel dosing method of Escherichia coli L-asparaginase (EC-Asnase) in children and adolescents with newly diagnosed acute lymphoblastic leukemia (ALL). Patients achieving complete remission (CR) on Dana-Farber Cancer Institute ALL Consortium Protocol 00-01 were eligible for random assignment to 1) dexamethasone or prednisone, administered as 5-day pulses, every 3 weeks, and 2) weekly EC-Asnase, administered as a 25,000 IU/m(2) fixed dose (FD) or individualized dose (ID) starting at 12,500-IU/m(2), adjusted every 3 weeks based on nadir serum asparaginase activity (NSAA) determinations. Between 2000 and 2004, 492 evaluable patients (ages 1 to 18 years) enrolled; 473 patients (96%) achieved CR. Four hundred eight patients (86%) participated in the corticosteroid randomization and 384 patients (81%) in the EC-Asnase randomization. With 4.9 years of median follow-up, dexamethasone was associated with superior 5-year event-free survival (EFS; 90% v 81% for prednisone; P = .01) but higher rates of infection (P = .03) and, in older children, higher cumulative incidence of osteonecrosis (P = .02) and fracture (P = .06). ID EC-Asnase had superior 5-year EFS (90% v 82% for FD; P = .04), but did not reduce the frequency of asparaginase-related toxicity. Multivariable analysis identified both dexamethasone and ID EC-Asnase as independent predictors of favorable EFS. There was no overall difference in skeletal toxicity by corticosteroid type; dexamethasone was associated with more infections and, in older children, increased incidence of osteonecrosis and fracture. There was no difference in asparaginase-related toxicity by EC-Asnase dosing method. Dexamethasone and ID EC-Asnase were each associated with superior EFS. Monitoring NSAA during treatment with EC-Asnase may be an effective strategy to improve outcome in pediatric ALL.


The effect of Stage II posterior tibial tendon dysfunction on deep compartment muscle strength: a new strength test.
The purpose of this study was to compare isometric subtalar inversion and forefoot adduction strength in subjects with Stage II posterior tibial tendon dysfunction (PTTD) to controls. Twenty four subjects with Stage II PTTD and fifteen matched controls volunteered for this study. A force transducer (Model SML-200, Interface, Scottsdale, AZ) was connected with a resistance plate and oscilloscope (TDS 410A, Tektronix, Beaverton, OR) to the foot. Via the oscilloscope, subjects were given feedback on the amount of force produced and muscle activation of the anterior tibialis (AT) muscle. Subjects were instructed to maintain a plantar flexion force while performing a maximal voluntary subtalar inversion and forefoot adduction effort. A two-way ANOVA model with the factors including, side (involved/uninvolved) and group (control/PTTD) was used. The PTTD group on the involved side showed significantly decreased subtalar inversion and foot adduction strength (0.70 +/- 0.24 N/Kg) compared to the uninvolved side (0.94 +/- 0.24 N/Kg) and controls (involved side = 0.99 +/- 0.24 N/Kg, uninvolved side = 0.97 +/- 0.21 N/Kg). The average AT activation was between 11% to 17% for both groups, however, considerable variability in subjects with PTTD. These data confirm a subtalar inversion and forefoot adduction strength deficit by 20% to 30% in subjects with Stage II PTTD. Although isolating the PT muscle is difficult, a test specific to subtalar inversion and forefoot adduction demonstrated the weakness in this population.


Optimising non-viral gene delivery in a tumour spheroid model.
Our current understanding of how the unique tumour microenvironment influences the efficacy of gene delivery is limited. The current investigation systematically examines the efficiency of several non-viral gene transfer agents to transfect multicellular tumour spheroids (MCTS), an in vitro model that displays a faithful three-dimensional (3D) representation of solid tumour tissue. Using a luciferase reporter assay, gene transfer to MCTS was optimised for 22 kDa linear and 25 kDa branched polyethyleneimine (PEI), the cationic lipids Lipofectamine(trade mark) and DCChol : DOPE, and the physical approach of tissue electroporation. Confocal microscopy was used to take optical tissue slices to identify the tissue localisation of green fluorescent protein (GFP) reporter gene expression and the distribution of fluorescently labelled complexes. A MCTS model of quiescent tumour regions was used to establish the influence of cellular proliferation status on gene transfer efficiency. Of the polyplexes tested, 22 kDa linear PEI provided optimal gene delivery, with gene expression peaking at 46 h. Despite being the optimal vector tested, PEI-mediated transfection was limited to cells at the MCTS periphery. Using fluorescent PEI, it was found that complexes could only penetrate the outer 3-5 proliferating cell layers of the MCTS, sparing the deeper quiescent cells. Gene delivery in an MCTS model comprised entirely of quiescent cells demonstrated that in addition to being inaccessible to the vector, quiescent tumour regions are inherently less susceptible to PEI-mediated transfection than proliferating regions. This 'resistance' to transfection observed in quiescent cells was overcome through the use of electroporation. Despite the improved efficacy of electroporation in quiescent tissue, the gene expression was still confined to the outer regions of MCTS. The results suggest that limited access to central regions of an MCTS remain a significant barrier to gene delivery. This data provides new insights into tumour-specific factors affecting non-viral gene transfer and highlights the difficulties in delivering genes to avascular tumour regions. The MCTS model is a useful system for the initial screening of future gene therapy strategies for solid tumours.


The contourlet transform: an efficient directional multiresolution image representation.
The limitations of commonly used separable extensions of one-dimensional transforms, such as the Fourier and wavelet transforms, in capturing the geometry of image edges are well known. In this paper, we pursue a "true" two-dimensional transform that can capture the intrinsic geometrical structure that is key in visual information. The main challenge in exploring geometry in images comes from the discrete nature of the data. Thus, unlike other approaches, such as curvelets, that first develop a transform in the continuous domain and then discretize for sampled data, our approach starts with a discrete-domain construction and then studies its convergence to an expansion in the continuous domain. Specifically, we construct a discrete-domain multiresolution and multidirection expansion using nonseparable filter banks, in much the same way that wavelets were derived from filter banks. This construction results in a flexible multiresolution, local, and directional image expansion using contour segments, and, thus, it is named the contourlet transform. The discrete contourlet transform has a fast iterated filter bank algorithm that requires an order N operations for N-pixel images. Furthermore, we establish a precise link between the developed filter bank and the associated continuous-domain contourlet expansion via a directional multiresolution analysis framework. We show that with parabolic scaling and sufficient directional vanishing moments, contourlets achieve the optimal approximation rate for piecewise smooth functions with discontinuities along twice continuously differentiable curves. Finally, we show some numerical experiments demonstrating the potential of contourlets in several image processing applications. Index Terms-Contourlets, contours, filter banks, geometric image processing, multidirection, multiresolution, sparse representation, wavelets.


Levels of Health Theory With the Example of a Case of Juvenile Rheumatoid Arthritis.
Contemporary medicine is in great need of a new patient health group classification that could be the basis for in-depth pathology assessments, disease development and prognosis, and the possibility of healing, as well as for possible complications of organism reactions to treatment processes. This classification is possible if founded on holistic approaches at the level of health assessment from the point of view of organism reactivity and resistance. Such a classification, assigning 12 levels and 4 health groups, exists in classical homeopathy. A new method for determining the group and level of health is shown in a case of juvenile rheumatoid arthritis of a generalized form in an 11-year-old girl, treated with classical homeopathy. The follow-up of the case is 18 years. The method allows the physician to assess the organism dynamics as a whole during the pathology development.


NIH conference. Airway inflammation.
Diseases characterized by airway inflammation, excessive airway secretion, and airway obstruction affect a substantial proportion of the population. These diseases include asthma, chronic bronchitis, bronchiectasis, and cystic fibrosis. Asthma and chronic bronchitis may affect 25 million persons in the United States. Much progress has been made in the last decade toward an understanding of the mechanisms underlying chronic airway inflammation; recent work has resulted in several new concepts of the initiation and maintenance of airway inflammation. Airway production of chemokines, cytokines, and growth factors in response to irritants, infectious agents, and inflammatory mediators may play an important role in the modulation of acute and chronic airway inflammation. Lipid mediators may be produced by resident airway cells and by inflammatory cells; production of these mediators may also be altered by inflammatory cytokines. Increased airway obstruction may be related to intercurrent viral respiratory infection and to the induction of airway inflammation and airway hyperreactivity that results from such infection. Furthermore, several models exist to explain the processes by which airway inflammation is perpetuated in diseases such as asthma and chronic bronchitis. These include neurogenic inflammation, the perpetuation of the acute inflammatory response, and cycles of airway epithelial cell-mediated and inflammatory cell-mediated recruitment and activation of inflammatory cells. An understanding of these mechanisms of airway inflammation may provide the clinician with new therapeutic approaches to the treatment of these common and chronic diseases.


Evolution and gene capture in ancient endogenous retroviruses - insights from the crocodilian genomes.
Crocodilians are thought to be hosts to a diverse and divergent complement of endogenous retroviruses (ERVs) but a comprehensive investigation is yet to be performed. The recent sequencing of three crocodilian genomes provides an opportunity for a more detailed and accurate representation of the ERV diversity that is present in these species. Here we investigate the diversity, distribution and evolution of ERVs from the genomes of three key crocodilian species, and outline the key processes driving crocodilian ERV proliferation and evolution. ERVs and ERV related sequences make up less than 2% of crocodilian genomes. We recovered and described 45 ERV groups within the three crocodilian genomes, many of which are species specific. We have also revealed a new class of ERV, ERV4, which appears to be common to crocodilians and turtles, and currently has no characterised exogenous counterpart. For the first time, we formally describe the characteristics of this ERV class and its classification relative to other recognised ERV and retroviral classes. This class shares some sequence similarity and sequence characteristics with ERV3, although it is phylogenetically distinct from the other ERV classes. We have also identified two instances of gene capture by crocodilian ERVs, one of which, the capture of a host KIT-ligand mRNA has occurred without the loss of an ERV domain. This study indicates that crocodilian ERVs comprise a wide variety of lineages, many of which appear to reflect ancient infections. In particular, ERV4 appears to have a limited host range, with current data suggesting that it is confined to crocodilians and some lineages of turtles. Also of interest are two ERV groups that demonstrate evidence of host gene capture. This study provides a framework to facilitate further studies into non-mammalian vertebrates and highlights the need for further studies into such species.


Antibacterial activity of isolated phenolic compounds from cranberry (Vaccinium macrocarpon) against Escherichia coli.
Phenolic compounds from a cranberry extract were isolated in order to assess their contribution to the antibacterial activity against uropathogenic strains of Escherichia coli (UPEC). With this purpose, a total of 25 fractions from a cranberry extract were isolated using semipreparative high performance liquid chromatography (HPLC) and characterized based on the results obtained by reversed-phase HPLC coupled to mass spectrometry detection. Then, the effects on UPEC surface hydrophobicity and biofilm formation of the cranberry extract as well as the purest fractions (a total of 13) were tested. As expected, the whole extract presented a powerful antibacterial activity against UPEC while the selected fractions presented a different behavior. Myricetin and quercitrin significantly decreased (p < 0.05) E. coli biofilm formation compared with the control, while dihydroferulic acid glucuronide, procyanidin A dimer, quercetin glucoside, myricetin and prodelphinidin B led to a significant decrease of the surface hydrophobicity compared with the control. The results suggest that apart from proanthocyanidins, other compounds, mainly flavonoids, can act against E. coli biofilm formation and also modify UPEC surface hydrophobicity in vitro, one of the first steps of adhesion.


Immune anaemias in patients with chronic lymphocytic leukaemia treated with fludarabine, cyclophosphamide and rituximab--incidence and predictors.
Immune anaemias (IA) [auto-immune haemolytic anaemia (AIHA) and pure red cell aplasia (PRCA)] are complications of chronic lymphocytic leukaemia (CLL). Fludarabine has been associated with AIHA, whereas both rituximab and cyclophosphamide have been used to treat this condition. Combining these agents with fludarabine may reduce the likelihood of AIHA. We report on the incidence, outcome and pretreatment predictors of IA in 300 patients treated with fludarabine, cyclophosphamide and rituximab (FCR). Nineteen patients (6.5%) developed IA [AIHA (5.8%), PRCA (0.7%)] on or after treatment with FCR. Most patients (82.4%) with AIHA had a negative direct antiglobulin test (DAT). Additional markers of haemolysis (indirect hyperbilirubinaemia, reticulocytosis, low haptoglobin and elevated lactate dehydrogenase levels) confirmed the presence of AIHA in these patients. The majority of patients responded to therapies including steroids, ciclosporin, i.v. immunoglobulin, etc. High pretreatment levels of beta-2 microglobulin predicted for development of IA. No haemolytic crisis was observed during FCR therapy in eight patients with AIHA prior to FCR. Thus, the incidence of IA among CLL patients treated with FCR was comparable with historical rates. The diagnosis of AIHA can be considered even if the DAT is negative. Pre-existing AIHA need not preclude front-line FCR therapy.


White blood cell and cell-free DNA analyses for detection of residual disease in gastric cancer.
Liquid biopsies are providing new opportunities for detection of residual disease in cell-free DNA (cfDNA) after surgery but may be confounded through identification of alterations arising from clonal hematopoiesis. Here, we identify circulating tumor-derived DNA (ctDNA) alterations through ultrasensitive targeted sequencing analyses of matched cfDNA and white blood cells from the same patient. We apply this approach to analyze samples from patients in the CRITICS trial, a phase III randomized controlled study of perioperative treatment in patients with operable gastric cancer. After filtering alterations from matched white blood cells, the presence of ctDNA predicts recurrence when analyzed within nine weeks after preoperative treatment and after surgery in patients eligible for multimodal treatment. These analyses provide a facile method for distinguishing ctDNA from other cfDNA alterations and highlight the utility of ctDNA as a predictive biomarker of patient outcome to perioperative cancer therapy and surgical resection in patients with gastric cancer.


Pseudorabies virus glycoprotein gIII is a major target antigen for murine and swine virus-specific cytotoxic T lymphocytes.
Pseudorabies virus (PrV) is the etiological agent of Aujeszky's disease, a disease that causes heavy economic losses in the swine industry. A rational approach to the generation of an effective vaccine against this virus requires an understanding of the immune response induced by it and of the role of the various viral antigens in inducing such a response. We have constructed mutants of PrV [strain PrV (Ka)] that differ from each other only in expression of the viral nonessential glycoproteins gI, gp63, gX, and gIII (i.e., are otherwise isogenic). These mutants were used to ascertain the importance of each of the nonessential glycoproteins in eliciting a PrV-specific cytotoxic T-lymphocyte (CTL) response in mice and pigs. Immunization of DBA/2 mice and pigs with a thymidine kinase-deficient (TK-) mutant of PrV elicits the formation of cytotoxic cells that specifically lyse syngeneic infected target cells. These PrV-specific cytolytic cells have the phenotype of major histocompatibility complex class I antigen-restricted CTLs. The relative number of CTLs specific for glycoproteins gI, gp63, gX, and gIII induced in mice vaccinated with a TK- mutant of PrV was ascertained by comparing their levels of cytotoxicity against syngeneic cells infected with either wild-type virus or gI-/gp63-, gX-, or gIII- virus deletion mutants. The PrV-specific CLTs were significantly less effective in lysing gIII(-)-infected targets than in lysing gI-/gp63-, gX-, or wild-type-infected targets. The in vitro secondary CTL response of lymphocytes obtained from either mice or pigs 6 or more weeks after immunization with a TK- mutant of PrV was also tested. Lymphocytes obtained from these animals were cultured with different glycoprotein-deficient mutants of PrV, and their cytolytic activities against wild-type-infected targets were ascertained. The importance of each of the nonessential viral glycoproteins in eliciting CTLs was assessed from the effectiveness of each of the virus mutants to stimulate the secondary anti-PrV CTL response. Cultures of both murine or swine lymphocytes that had been stimulated with gIII- virus contained only approximately half as many lytic units as did those stimulated with either wild-type virus, a gX- virus mutant, or a gI-/gp63- virus mutant. Thus, a large proportion of the PrV-specific CTLs that are induced by immunization with PrV of both mice and pigs are directed against gIII. Furthermore, glycoproteins gI, gp63, and gX play at most a minor role in the CTL response of these animals to PrV.


Preparation of surfactant-free nanoparticles of methacrylic acid copolymers used for film coating.
The aim of the present study was to prepare surfactant-free pseudolatexes of various methacrylic acid copolymers. These aqueous colloidal dispersions of polymeric materials for oral administration are intended for film coating of solid dosage forms or for direct manufacturing of nanoparticles. Nanoparticulate dispersions were produced by an emulsification-diffusion method involving the use of partially water-miscible solvents and the mutual saturation of the aqueous and organic phases prior to the emulsification in order to reduce the initial thermodynamic instability of the emulsion. Because of the self-emulsifying properties of the methacrylic acid copolymers, it was possible to prepare aqueous dispersions of colloidal size containing up to 30% wt/vol of Eudragit RL, RS, and E using 2-butanone or methyl acetate as partially water-miscible solvents, but without any surfactant. However, in the case of the cationic Eudragit E, protonation of the tertiary amine groups by acidification of the aqueous phase was necessary to improve the emulsion stability in the absence of surfactant and subsequently to prevent droplet coalescence during evaporation. In addition, a pseudolatex of Eudragit E was used to validate the coating properties of the formulation for solid dosage forms. Film-coated tablets of quinidine sulfate showed a transparent glossy continuous film that was firmly attached to the tablet. The dissolution profile of quinidine sulfate from the tablets coated with the Eudragit E pseudolatex was comparable to that of tablets coated with an acetonic solution of Eudragit E. Furthermore, both types of coating ensured similar taste masking. The emulsification-evaporation method used was shown to be appropriate for the preparation of surfactant-free colloidal dispersions of the 3 types of preformed methacrylic acid copolymers; the dispersions can subsequently be used for film coating of solid dosage forms.


[Attitudes of personnel monitoring intra-hospital adverse events in Colombia].
Detecting adverse events (AE) is part of managing hospitalised patients' safety. Suitable AE monitoring is affected by many factors regarding any particular institution and its workers. This article was aimed at describing the difficulties emerging from identifying and evaluating events and suggested interventions from the workers' viewpoint. Qualitative research. A focus group was formed with key informants from previous research entitled 'The incidence and avoidability of adverse events in three institutions in Colombia, 2006.' A conceptual framework was constructed based on publications pertaining to AEs and health-providing institutions; summaries were made by topic regarding the content of the focus group's work by systematising, categorising and readjusting the data. The triangulation method was used for guar-anteeing its credibility, transferability, reliability and that it could be confirmed. It is commonly thought that monitoring AE only consists of taking inventories related to negative work consequences into account; this opinion has been determined by prior organisational culture. Strategies used for increasing AE reporting were: intrapersonal work, raising awareness that nobody is exempt from being involved in an adverse event and encouraging administrative support for resolving deficiencies. The prospective monitoring method becomes hampered when applying it to services involving a large volume of information. A tendency was observed for specialist committees to underestimate an event's association. Heath workers, support personnel and management must have mutual confidence and adopt team-work so that future AE may be prevented.


Dental anxiety and fear: relationship with oral health behavior in a Turkish population.
The aim of this study was to assess fear and anxiety in dental patients. Five hundred patients were evaluated using the Modified Dental Anxiety Scale and the Dental Fear Scale, along with a questionnaire. Oral health status was assessed using the Decayed, Missing, and Filled Teeth (DMFT)/Decayed, Missing, and Filled Surfaces (DMFS) index. Statistic al analysis was performed (P < .05). Sex significantly affected dental anxiety (P < .05), and sex, marital status, having children, and time elapsed since last visit to clinician affected dental fear (P < .05). DMFT/DMFS scores were not related to dental anxiety or fear (P > .05). Female sex alone was a significant predictor of dental anxiety; female sex, adulthood, marriage, having children, and time passed since last visit to a clinician are significant predictors of fear.


TMEM70 deficiency: long-term outcome of 48 patients.
TMEM70 deficiency is the most common nuclear-encoded defect affecting the ATP synthase. In this multicentre retrospective study we characterise the natural history of the disease, treatment and outcome in 48 patients with mutations in TMEM70. Eleven centers from eight European countries, Turkey and Israel participated. All 27 Roma and eight non-Roma patients were homozygous for the common mutation c.317-2A > G. Five patients were compound heterozygotes for the common mutation and mutations c.470 T > A, c.628A > C, c.118_119insGT or c.251delC. Six Arab Muslims and two Turkish patients were homozygous for mutations c.238C > T, c.316 + 1G > T, c.336 T > A, c.578_579delCA, c.535C > T, c.359delC. Age of onset was neonatal in 41 patients, infantile in six cases and two years in one child. The most frequent symptoms at onset were poor feeding, hypotonia, lethargy, respiratory and heart failure, accompanied by lactic acidosis, 3-methylglutaconic aciduria and hyperammonaemia. Symptoms further included: developmental delay (98%), hypotonia (95%), faltering growth (94%), short stature (89%), non-progressive cardiomyopathy (89%), microcephaly (71%), facial dysmorphism (66%), hypospadias (50% of the males), persistent pulmonary hypertension of the newborn (22%) and Wolff-Parkinson-White syndrome (13%). One or more acute metabolic crises occurred in 24 surviving children, frequently followed by developmental regression. Hyperammonaemic episodes responded well to infusion with glucose and lipid emulsion, and ammonia scavengers or haemodiafiltration. Ten-year survival was 63%, importantly for prognostication, no child died after the age of five years. TMEM70 deficiency is a panethnic, multisystemic disease with variable outcome depending mainly on adequate management of hyperammonaemic crises in the neonatal period and early childhood.


Pulmonary gas conducting interstitial pathway.
In spite of the growing efforts oriented towards revealing different aspects of emphysema, the persistence of the emphysematous or emphysema-like changes (ELCs) is not explored yet in the open literature. In this study we demonstrate the persistence of an ELC for 22 years in a spontaneous pneumothorax (SP) patient which indicates a hitherto unknown gas supply to the ELC. For this purpose we used high resolution computed tomography (HRCT) images processed into three-dimensional (3D) geometry. By the same token, not only a long persistence but also the volume increase of this ELC between 2002 and 2010 was demonstrated. The 3D geometry visualized an aerated interstitial structure between the sites of supposed gas leakage at the wall of the third generation airways and the ELC. This potential gas conducting interstitial pathway is not a continuation and has neither the form nor the structure of a bronchus. The finding suggests that in this patient the intrabronchial gas passes through the bronchial wall and via a gas conducting interstitial pathway reaches the ELC. Despite the availability of the presently employed techniques for at least 15 years, such case and phenomenon have not been described previously. The retrieval of the patient suggests that the findings could be relevant for a considerable proportion of the population.


Carcinogenesis Bioassay of Vinylidene Chloride (CAS No. 75-35-4) in F344 Rats and B6C3F1 Mice (Gavage Study).
A subchronic and a chronic carcinogenesis study of vinylidene chloride (99% pure), a widely used chemical intermediate and monomer, were conducted in F344/N rats and B6C3F1/N mice. In subchronic studies, groups of 10 rats and 10 mice of either sex were administered vinylidene chloride in corn oil by gavage five times per week at 0, 5, 15, 50, 100, or 250 mg/kg body weight for 13 weeks. At the end of this study, representative tissues from these animals were subjected to histopathological examination. The liver was identified as a target organ for vinylidene chloride toxicity. In the 104-week chronic exposure study, conducted primarily to determine possible carcinogenic potential of vinylidene chloride by the oral route, 50 F344/N rats and 50 B6C3F1/N mice of either sex were gavaged with vinylidene chloride suspended in corn oil at dose levels of 1 or 5 mg/kg (rats) and 2 or 10 mg/kg (mice). Groups of 50 rats and 50 mice of either sex received corn oil alone and served as vehicle controls. Throughout most of the study, mean body weights of the dosed rats of either sex and high-dose female mice were slightly lower than those of the controls. The absence of compound-related effects on survival or clinical signs suggests that the rats and mice of either sex could have tolerated higher doses. While no significant differences in survival were observed for any group of rats, 12 control and 10 low-dose males were killed accidentally during week 82; this may have compromised the sensitivity of the male rats study. The results of histopathological examination indicated an increased incidence of necrosis of the liver in high-dose male and low-dose female mice and chronic renal inflammation in high-dose rats of either sex. The only observed significant (P>0.05) increase in tumor incidence occurred in low-dose female mice: lymphoma (2/48, 9/49, 6/50) and lymphoma or leukemia (7/48, 15/49, 7/50). These increases were not considered to be related to vinylidene chloride administration because similar effects were not found in the high-dose female mice or in male mice or rats. Under the conditions of this bioassay, vinylidene chloride administered by gavage was not carcinogenic for F344/N rats or B6C3F1/N mice of either sex. However, since the use of a maximum tolerated dose in this study has not been clearly demonstrated and since previously reported studies have shown that carcinogenicity is associated with inhalation exposure to vinylidene chloride, this study should not be taken as proof that the chemical is not a carcinogen. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonyms: 1,1-dichloroethylene; VDC; 1,1-DCE


S6K1 is a multifaceted regulator of Mdm2 that connects nutrient status and DNA damage response.
p53 mediates DNA damage-induced cell-cycle arrest, apoptosis, or senescence, and it is controlled by Mdm2, which mainly ubiquitinates p53 in the nucleus and promotes p53 nuclear export and degradation. By searching for the kinases responsible for Mdm2 S163 phosphorylation under genotoxic stress, we identified S6K1 as a multifaceted regulator of Mdm2. DNA damage activates mTOR-S6K1 through p38alpha MAPK. The activated S6K1 forms a tighter complex with Mdm2, inhibits Mdm2-mediated p53 ubiquitination, and promotes p53 induction, in addition to phosphorylating Mdm2 on S163. Deactivation of mTOR-S6K1 signalling leads to Mdm2 nuclear translocation, which is facilitated by S163 phosphorylation, a reduction in p53 induction, and an alteration in p53-dependent cell death. These findings thus establish mTOR-S6K1 as a novel regulator of p53 in DNA damage response and likely in tumorigenesis. S6K1-Mdm2 interaction presents a route for cells to incorporate the metabolic/energy cues into DNA damage response and links the aging-controlling Mdm2-p53 and mTOR-S6K pathways.


Effect of hydration on exercise-induced growth hormone response.
Growth hormone (GH) has demonstrated water-retaining effects in subjects at rest, whereas other research has indicated that GH may stimulate sweating. Thus, the aim of this study was to investigate the effect of fluid intake on the exercise-induced GH response. Seven healthy male volunteers (age: 27.4+/-1.3 years, weight: 74.5+/-1.1 kg, height: 179.3+/-2.3 cm) performed a 40-min submaximal rectangular cycling exercise in two different sessions. The first session (Session 1) was performed without water intake, and the second (Session 2) involved the ingestion of spring water (four intakes) corresponding to the volume of water lost during the first session. In session 1, the water loss was 568+/-32 ml. In Session 2, the volume of water loss was not significantly different from the volume of fluid intake (524+/-16 versus 568+/-32 ml respectively). The decrease in plasma volume was significantly reduced in Session 2 (-6.69+/-1.59% versus -11.3+/-1.89%; P<0.05). In Session 1, the GH concentration was significantly lower than that during Session 2 after 25 min (3.04+/-1.05 versus 5.26+/-1.81; P<0.05) and after 40 min (13.7+/-3.55 versus 17.60+/-4.14 ng/ml; P<0.05) of exercise. The total GH response was significantly lower in Session 1 than in Session 2 (136.6+/-39.2 versus 202.4+/-58.9 ng/ml x min; P<0.05). We conclude that the exercise-induced GH response decreases when exercise is performed without fluid intake.


Comparison of P388D1 mouse macrophage cell line and human monocytes for assay of dengue-2 infection-enhancing antibodies.
Tissue culture-adapted dengue 2 virus (DEN 2), strain 16681, exhibits antibody-dependent enhancement of infection (ADE) in P388D1 cells, a mouse macrophage-like cell line. ADE is dependent upon maintaining DEN 2 multiplicity of infection at between 0.1 and 0.001, and can be simply measured in multi-well plastic plates. The assay uses either trypsinized or non-trypsinized P388D1 cells at 5 x 10(5) cells per ml, an appropriate dilution of DEN 2 virus, and a source of antibody, and is most conveniently performed without further washing of stationary cultures, which are incubated in 5% CO2. Trypsinization of P388D1 cells prior to the addition of virus-serum mixtures reduced infection in control cultures thus increasing ADE. When cells were washed after incubation of virus-serum mixtures for 1 hour, a paradoxical increase of infection in cultures exposed to virus plus normal serum was noted, which reduced the sensitivity of the ADE assay. Using human cord blood sera, ADE titers measured in human monocytes and P388D1 cells were closely similar. This convenient and economical assay will facilitate large scale biological and epidemiological studies of dengue virus enhancing antibodies.


Effect of intravenous β-2 agonist treatment on clinical outcomes in acute respiratory distress syndrome (BALTI-2): a multicentre, randomised controlled trial.
In a previous randomised controlled phase 2 trial, intravenous infusion of salbutamol for up to 7 days in patients with acute respiratory distress syndrome (ARDS) reduced extravascular lung water and plateau airway pressure. We assessed the effects of this intervention on mortality in patients with ARDS. We did a multicentre, placebo-controlled, parallel-group, randomised trial at 46 UK intensive-care units between December, 2006, and March, 2010. Intubated and mechanically ventilated patients (aged ≥16 years) within 72 h of ARDS onset were randomly assigned to receive either salbutamol (15 μg/kg ideal bodyweight per h) or placebo for up to 7 days. Randomisation was done by a central telephone or web-based randomisation service with minmisation by centre, pressure of arterial oxygen to fractional inspired oxygen concentration (PaO(2)/F(I)O(2)) ratio, and age. All participants, caregivers, and investigators were masked to group allocation. The primary outcome was death within 28 days of randomisation. Analysis was by intention-to-treat. This trial is registered, ISRCTN38366450 and EudraCT number 2006-002647-86. We randomly assigned 162 patients to the salbutamol group and 164 to the placebo group. One patient in each group withdrew consent. Recruitment was stopped after the second interim analysis because of safety concerns. Salbutamol increased 28-day mortality (55 [34%] of 161 patients died in the salbutamol group vs 38 (23%) of 163 in the placebo group; risk ratio [RR] 1·47, 95% CI 1·03-2·08). Treatment with intravenous salbutamol early in the course of ARDS was poorly tolerated. Treatment is unlikely to be beneficial, and could worsen outcomes. Routine use of β-2 agonist treatment in ventilated patients with this disorder cannot be recommended. UK Medical Research Council, UK Department of Health, UK Intensive Care Foundation.


Chemical nature of nitric oxide storage forms in rat vascular tissue.
Endothelial NO production results in local formation of adducts that may act as storage forms of NO. Because little is known about their chemical nature, concentrations, and possible role in vascular biology, we sought to characterize those species basally present in rat aorta, using two independent approaches. In the first approach, tissue homogenates were analyzed by using chemiluminescence- and ion-chromatography-based techniques that allow trace-level quantification of NO-related compounds in complex biological matrices. In the second approach, NO stores were characterized by their ability to release NO when illuminated with light and subsequently relax vascular smooth muscle (photorelaxation). The latter included a careful assessment of action spectra for photorelaxation, taking into account the light-scattering properties of the tissue and the storage depletion rates induced by exposure to controlled levels of light. Biochemical analyses revealed that aortic tissues contained 10 +/- 1 microM nitrite, 42 +/- 7 microM nitrate, 40 +/- 6 nM S-nitroso, and 33 +/- 6 nM N-nitroso compounds (n = 4-8). The functional data obtained suggest that the NO photolytically released in the tissue originated from species with photophysical properties similar to those reported for low-molecular-weight S-nitrosothiols, as well as from nitrite. The relative contribution of these potential NO stores to the extent of photorelaxation was consistent with their concentrations detected biochemically in vascular tissue when their photoactivity was taken into account. We conclude that intravascular nitroso species and nitrite both have the potential to release physiologically relevant quantities of NO independent of enzymatic control by NO synthase.


[Certain clinico-anatomic features of large and giant arterial aneurysms of cerebral vessels].
On the basis of examining 72 patients 14 of whom later died it has been concluded that the most substantial differences of large and giant aneurysms of cerebral arteries from small ones consist in development of focal neurological symptoms and intracranial hypertension: this is due to both compression of the brain structures and disturbance of the cerebral circulation caused by aneurysms more than 2 cm long. The intracranial hypertension is the most frequent in aneurysms over 3 cm long, especially, if those aneurysms are localized in the vicinity of cerebral liquor ducts. Patients with arterial aneurysms exceeding 2 cm develop hemorrhagic complications in 2/3 of the cases. While using modern treatment methods (stereotaxic and intravascular) one should take into account the peculiarities of the structure of large and giant aneurysms in the brain.


A Protective Vaccine against Chlamydia Genital Infection Using Vault Nanoparticles without an Added Adjuvant.
Chlamydia trachomatis genital infection is the most common sexually transmitted bacterial disease, causing a significant burden to females due to reproductive dysfunction. Intensive screening and antibiotic treatment are unable to completely prevent female reproductive dysfunction, thus, efforts have become focused on developing a vaccine. A major impediment is identifying a safe and effective adjuvant which induces cluster of differentiation 4 (CD4) cells with attributes capable of halting genital infection and inflammation. Previously, we described a natural nanocapsule called the vault which was engineered to contain major outer membrane protein (MOMP) and was an effective vaccine which significantly reduced early infection and favored development of a cellular immune response in a mouse model. In the current study, we used another chlamydial antigen, a polymorphic membrane protein G-1 (PmpG) peptide, to track antigen-specific cells and evaluate, in depth, the vault vaccine for its protective capacity in the absence of an added adjuvant. We found PmpG-vault immunized mice significantly reduced the genital bacterial burden and histopathologic parameters of inflammation following a C. muridarum challenge. Immunization boosted antigen-specific CD4 cells with a multiple cytokine secretion pattern and reduced the number of inflammatory cells in the genital tract making the vault vaccine platform safe and effective for chlamydial genital infection. We conclude that vaccination with a Chlamydia-vault vaccine boosts antigen-specific immunities that are effective at eradicating infection and preventing reproductive tract inflammation.


Pregabalin attenuates excitotoxicity in diabetes.
Diabetes can exacerbate seizures and worsen seizure-related brain damage. In the present study, we aimed to determine whether the standard antiepileptic drug pregabalin (PGB) protects against pilocarpine-induced seizures and excitotoxicity in diabetes. Adult male Sprague-Dawley rats were divided into either a streptozotocin (STZ)-induced diabetes group or a normal saline (NS) group. Both groups were further divided into subgroups that were treated intravenously with either PGB (15 mg/kg) or a vehicle; all groups were treated with subcutaneous pilocarpine (60 mg/kg) to induce seizures. To evaluate spontaneous recurrent seizures (SRS), PGB-pretreated rats were fed rat chow containing oral PGB (450 mg) for 28 consecutive days; vehicle-pretreated rats were fed regular chow. SRS frequency was monitored for 2 weeks from post-status epilepticus day 15. We evaluated both acute neuronal loss and chronic mossy fiber sprouting in the CA3 area. In addition, we performed patch clamp recordings to study evoked excitatory postsynaptic currents (eEPSCs) in hippocampal CA1 neurons for both vehicle-treated rats with SRS. Finally, we used an RNA interference knockdown method for Kir6.2 in a hippocampal cell line to evaluate PGB's effects in the presence of high-dose ATP. We found that compared to vehicle-treated rats, PGB-treated rats showed less severe acute seizure activity, reduced acute neuronal loss, and chronic mossy fiber sprouting. In the vehicle-treated STZ rats, eEPSC amplitude was significantly lower after PGB administration, but glibenclamide reversed this effect. The RNA interference study confirmed that PGB could counteract the ATP-sensitive potassium channel (KATP)-closing effect of high-dose ATP. By opening KATP, PGB protects against neuronal excitotoxicity, and is therefore a potential antiepileptogenic in diabetes. These findings might help develop a clinical algorithm for treating patients with epilepsy and comorbid metabolic disorders.


Putative glycoprotein and glycolipid polymorphonuclear leukocyte receptors for the Actinomyces naeslundii WVU45 fimbrial lectin.
Recognition of receptors on sialidase-treated polymorphonuclear leukocytes (PMNs) by the Gal/GalNAc lectin associated with the type 2 fimbriae of certain strains of actinomyces results in activation of the PMNs, phagocytosis, and destruction of the bacteria. In the present study, plant lectins were utilized as probes to identify putative PMN receptors for the actinomyces lectin. The Gal-reactive lectin from Ricinus communis (RCAI), the Gal/GalNAc-reactive lectins from R. communis (RCAII) and Bauhinia purpurea (BPA), as well as the Gal beta 1-3GalNAc-specific lectins from Arachis hypogaea (PNA) and Agaricus bisporus (ABA) inhibited killing of Actinomyces naeslundii WVU45 by sialidase-treated PMNs. These five lectins detected a 130-kDa surface-labeled glycoprotein on nitrocellulose transfers of PMN extracts separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This glycoprotein was revealed only after treatment of the transfers with sialidase, a condition analogous to the sialidase dependence of the lectin-mediated biological responses of the PMNs to the actinomyces. The mannose-reactive lectin concanavalin A did not inhibit killing of the actinomyces and failed to detect the 130-kDa glycoprotein but did block PMN-dependent killing of Escherichia coli B, a bacterium that possesses mannose-sensitive fimbriae. Therefore, the PMN glycoprotein receptor for A. naeslundii is clearly distinct from those recognized by E. coli. Two major putative glycolipid receptors were also identified by actinomyces and RCAI overlays on sialidase-treated thin-layer chromatograms of PMN gangliosides. Thus, both a 130-kDa glycoprotein and certain gangliosides are implicated in the attachment of the actinomyces to PMNs.


NF beta A, a factor required for maximal interleukin-1 beta gene expression is identical to the ets family member PU.1. Evidence for structural alteration following LPS activation.
We have previously identified and characterized the macrophage-, neutrophil- and B cell-specific nuclear factor beta A (NF beta A), which is involved in transcriptional regulation of the interleukin-1 beta (IL-1 beta) gene. NF beta A binds to a highly conserved sequence element located 6 bp upstream of the TATA motif within the IL-1 beta promoter and is required for maximal expression of the IL-1 beta gene. Here we show that NF beta A is identical to the previously identified ets gene family member PU.1. The NF beta A binding element shares 100% sequence identity with a novel PU.1 binding element recently found in the immunoglobulin J-chain promoter. Methylation interference DNA footprinting data demonstrated that NF beta A and PU.1 make identical protein/DNA contacts. In vitro synthesized PU.1 possesses a mobility and binding specificity identical to NF beta A as determined by electrophoretic mobility shift analysis (EMSA). Antisera directed against amino acids 39-55 of PU.1 recognizes NF beta A in a manner indistinguishable from PU.1 in EMSA 'supershift' studies. NF beta A and PU.1 also possess similar protein structure as determined by proteolytic clipping bandshift analysis. Furthermore, we show that PU.1 is able to transactivate an NF beta A-dependent promoter when co-transfected into HeLa cells which lack PU.1/NF beta A. EMSA studies using recombinant TATA binding protein (TBP) and PU.1 suggest that PU.1 may induce assembly of a distinct TBP-dependent complex on the IL-1 beta promoter. Finally, immunohistochemical confocal laser scanning microscopy studies suggest that LPS stimulation of RAW macrophages induces a structural change in the N-terminal transcriptional activation domain of PU.1.


Macrophages and their interactions with oncolytic viruses.
Macrophages are a highly plastic cell type and exhibit a range of defensive and regulatory functions in normal physiology. Phagocytic macrophages play an important role in defending against virus infection and they provide an important barrier that can limit the delivery of therapeutic viruses from the injection to the tumour. Within tumours, macrophages generally adopt an immunosuppressive phenotype and are associated with poor clinical prognosis. However their plasticity also provides the opportunity for therapeutic 're-education' of tumour-associated macrophages (TAMs) to adopt an active anticancer role. Oncolytic viruses present the possibility for non-specific stimulation of TAMs, and also the option for tumour-targeted expression of cytokines chosen specifically to modulate macrophage activation.


Effects of fibrin glue on hemostasis.
In order to decrease complications following incomplete hemostasis, we tried to make a safe, efficient and highly concentrated fibrin glue by thawing single-donor fresh frozen plasma. Using a unique animal model, in which arterial bleeding was created, fibrin glue and some related hemostatic agents were tested to evaluate their hemostatic effectiveness. The results demonstrated that: 1) the concomitant use of cryoprecipitate-thrombin tissue glue with adjuvant (aprotinin or calcium chloride) had a better hemostatic effect than the use of cryoprecipitate-thrombin tissue glue alone (p < 0.05); 2) impregnation of fibrin glue with a suitable vehicle was advisable to accelerate the coagulation plug formation and to enhance the mechanical strength of the adhesive plug; 3) Gelform, used as a vehicle to hold the fibrin glue, had a more efficient hemostatic effect than gauze, collagen fleece and Surgicel (p < 0.05); 4) systemic heparinization attenuated the effectiveness of the hemostatic agents and aggravated the bleeding problem, but a low hematocrit level did not; 5) fibrin glue had its own limitations, especially under systemic heparinization, on hemostatic effectiveness in a high-pressure system. Understanding the characteristics of fibrin glue, as mentioned above, definitely improved the hemostatic effectiveness of the glue, especially after failure of the usual methods of controlling bleeding.


Determinants of early recurrence of cerebral infarction. The Stroke Data Bank.
We studied 1,273 patients with ischemic cerebral infarction who were entered into the Stroke Data Bank, a prospective, observational study involving four university hospitals and the Biometry and Field Studies Branch of the National Institute of Neurological Disorders and Stroke. Forty patients had noniatrogenic recurrent stroke within 30 days after the index cerebral infarction. Using life tables, the 30-day cumulative +/- SE risk of early recurrence for all infarctions was 3.3 +/- 0.4%. The risk of early recurrence was greatest for atherothrombotic infarction (7.9 +/- 2.2%, eight of 113 patients) and least for lacunar infarction (2.2 +/- 1.2%, eight of 337 patients). Both cardioembolic infarction (4.3 +/- 0.9%, 10 of 246 patients) and infarction of undetermined cause (3.0 +/- 0.5%, 14 of 508 patients) had intermediate risks. History of hypertension and diabetes mellitus, as well as diastolic hypertension and elevated blood sugar concentration at admission, were associated with early recurrence. Logistic regression analysis estimated the risk of early recurrence to be 8.56% in those with coexisting hypertension and a glucose concentration of 300 mg/dl versus 0.77% in the absence of these two abnormalities. Early recurrence was associated with longer median duration of initial hospital stay (27 vs. 14 days) and a higher 30-day case-fatality rate (20% vs. 7.4%). Increased weakness scores were associated with early recurrent stroke. Identification of the determinants of early recurrent stroke may lead to better secondary prevention and may help select high-risk patients for further study.


Multimodal mimicry of hosts in a radiation of parasitic finches.
Brood parasites use the parental care of others to raise their young and sometimes employ mimicry to dupe their hosts. The brood-parasitic finches of the genus Vidua are a textbook example of the role of imprinting in sympatric speciation. Sympatric speciation is thought to occur in Vidua because their mating traits and host preferences are strongly influenced by their early host environment. However, this alone may not be sufficient to isolate parasite lineages, and divergent ecological adaptations may also be required to prevent hybridization collapsing incipient species. Using pattern recognition software and classification models, we provide quantitative evidence that Vidua exhibit specialist mimicry of their grassfinch hosts, matching the patterns, colors and sounds of their respective host's nestlings. We also provide qualitative evidence of mimicry in postural components of Vidua begging. Quantitative comparisons reveal small discrepancies between parasite and host phenotypes, with parasites sometimes exaggerating their host's traits. Our results support the hypothesis that behavioral imprinting on hosts has not only enabled the origin of new Vidua species, but also set the stage for the evolution of host-specific, ecological adaptations.


Quantitative assessment of hip morphology to enhance the identification of hip dysplasia in German Shepherd Dogs.
To develop quantitative measures that, when combined with the Fédération Cynologique Internationale (FCI) score, would potentially enhance the accuracy of the scoring process. 153 client-owned purebred German Shepherd Dogs with normal and near normal (71 dogs) and dysplastic coxofemoral joint (82 dogs). Center edge (CE) angle, Norberg angle (NA), indexes of dorsal acetabular femoral head (AFH) coverage width and area, acetabular index angle, and inclination angle were determined. We also investigated the correlation between selected variables. Coxofemoral joints were classified into normal, near normal, and mildly, moderately, and severely dysplastic joints based on the morphometric criteria previously established by the conventional FCI scoring. Variables were compared among the 5 groups using ANOVA. Linear relationships were determined using Spearman correlation coefficients. All radiographic measurements differed significantly (P < .0001) among the 5 assigned groups (normal, near normal, mildly dysplastic, moderately dysplastic, and severely dysplastic hip joints). NA was the only measure that differed significantly (P ≤ .03) between the 5 assigned groups. Positive correlations were identified between Norberg and CE angles (rs = 0.93), between width and area indexes of dorsal AFH coverage (rs = 0.92), and between the measurement techniques utilized to assess lateral versus dorsal AFH coverage (rs ≥ 0.65). Evaluation of lateral and dorsal AFH coverage may help to refine the scoring system used to select German Shepherd Dogs for breeding. German Shepherd Dogs with NA < 103°, CE angle < 20.8°, dorsal AFH coverage width index ≤ 49%, and/or dorsal AFH coverage area index ≤ 51% should be considered to have mild, moderate, or severe hip dysplasia and are therefore not good candidates for breeding. Borderline values between near normal and mildly dysplastic joints should be reevaluated.


Abraxas and RAP80 form a BRCA1 protein complex required for the DNA damage response.
The BRCT repeats of the breast and ovarian cancer predisposition protein BRCA1 are essential for tumor suppression. Phosphopeptide affinity proteomic analysis identified a protein, Abraxas, that directly binds the BRCA1 BRCT repeats through a phospho-Ser-X-X-Phe motif. Abraxas binds BRCA1 to the mutual exclusion of BACH1 (BRCA1-associated C-terminal helicase) and CtIP (CtBP-interacting protein), forming a third type of BRCA1 complex. Abraxas recruits the ubiquitin-interacting motif (UIM)-containing protein RAP80 to BRCA1. Both Abraxas and RAP80 were required for DNA damage resistance, G(2)-M checkpoint control, and DNA repair. RAP80 was required for optimal accumulation of BRCA1 on damaged DNA (foci) in response to ionizing radiation, and the UIM domains alone were capable of foci formation. The RAP80-Abraxas complex may help recruit BRCA1 to DNA damage sites in part through recognition of ubiquitinated proteins.


A Near-Global Slowing of Background Activity and Epileptic Discharges in Children With Mild to Moderately Symptomatic COVID-19 Infection: An Electro-Neurophysiological Study.
Background. To assess the functional involvement of the central nervous system (CNS) via quantitative electroencephalography (EEG) analysis in children with mild to moderate COVID-19 infection who were otherwise previously healthy children. Methods. This prospective, case-control study was conducted between June and September 2020. Sleep EEG records of at least 40 min were planned for children who tested positive for COVID-19 using real-time PCR analysis and within 4-6 months post-recovery. All of the EEG analyses in this study were performed on an Ubuntu 20.04.2 LTS Operating System with the developed software using Python 3.7.6. The quantitative analysis of the epileptic discharges within the EEG records was performed using random forest after elimination of the artifacts with a model training accuracy of 98% for each sample data point. The frequency analysis was performed using the Welch method. Results. Among the age and sex-matched groups, the global mean frequency was significantly lower among the COVID-19 patients, with a P-value of 0.004. The spike slow-wave and sharp slow-wave indices were significantly higher in the patients when compared to the controls. The mean frequency values were significantly lower in almost all of the electrodes recording the frontal, central, and occipital areas. For the temporal and parietal areas, those significantly low mean frequencies were limited to the right hemisphere. Conclusion. A near-global involvement of background activity with decreased frequency, in addition to epileptic discharges, was recorded in mild to moderately COVID-19 infected children post-infection.


A Minimal DNA Methylation Signature in Oral Tongue Squamous Cell Carcinoma Links Altered Methylation with Tumor Attributes.
Oral tongue squamous cell carcinomas (OTSCC) are a homogenous group of aggressive tumors in the head and neck region that spread early to lymph nodes and have a higher incidence of regional failure. In addition, there is a rising incidence of oral tongue cancer in younger populations. Studies on functional DNA methylation changes linked with altered gene expression are critical for understanding the mechanisms underlying tumor development and metastasis. Such studies also provide important insight into biomarkers linked with viral infection, tumor metastasis, and patient survival in OTSCC. Therefore, we performed genome-wide methylation analysis of tumors (N = 52) and correlated altered methylation with differential gene expression. The minimal tumor-specific DNA 5-methylcytosine signature identified genes near 16 different differentially methylated regions, which were validated using genomic data from The Cancer Genome Atlas cohort. In our cohort, hypermethylation of MIR10B was significantly associated with the differential expression of its target genes NR4A3 and BCL2L11 (P = 0.0125 and P = 0.014, respectively), which was inversely correlated with disease-free survival (P = 9E-15 and P = 2E-15, respectively) in patients. Finally, differential methylation in FUT3, TRIM5, TSPAN7, MAP3K8, RPS6KA2, SLC9A9, and NPAS3 genes was found to be predictive of certain clinical and epidemiologic parameters. This study reveals a functional minimal methylation profile in oral tongue tumors with associated risk habits, clinical, and epidemiologic outcomes. In addition, NR4A3 downregulation and correlation with patient survival suggests a potential target for therapeutic intervention in oral tongue tumors. Data from the current study are deposited in the NCBI Geo database (accession number GSE75540). Mol Cancer Res; 14(9); 805-19. ©2016 AACR.


Lectin switching during dengue virus infection.
Dengue virus receptors are relatively poorly characterized, but there has been recent interest in 2 C-type lectin molecules, dendritic cell-specific intercellular adhesion molecule 3 (ICAM-3)-grabbing nonintegrin (DC-SIGN) and its close homologue liver/lymph node-specific ICAM-3-grabbing integrin (L-SIGN), which can both bind dengue and promote infection. In this report we have studied the interaction of dengue viruses produced in insect cells, tumor cell lines, and primary human dendritic cells (DCs) with DC-SIGN and L-SIGN. Virus produced in primary DCs is unable to interact with DC-SIGN but remains infectious for L-SIGN-expressing cells. Skin-resident DCs may thus be a site of initial infection by insect-produced virus, but DCs will likely not participate in large-scale virus replication during dengue infection. These results reveal that differential glycosylation of dengue virus envelope protein is highly dependent on cell state and suggest that studies of virus tropism using virus prepared in insect cells or tumor cell lines should be interpreted with caution.


Osteochondral Allograft Transplantation in the Patellofemoral Joint: A Systematic Review.
The initial focus of cartilage restoration algorithms has been on the femur; however, the patellofemoral compartment accounts for 20% to 30% of significant symptomatic chondral pathologies. While patellofemoral compartment treatment involves a completely unique subset of comorbidities, with a comprehensive and thoughtful approach many patients may benefit from osteochondral allograft treatment. To perform a systematic review of clinical outcomes and failure rates after osteochondral allograft transplantation (OCA) of the patellofemoral joint at a minimum 18-month follow-up. Systematic review; Level of evidence, 4. A systematic review of the literature regarding the existing evidence for clinical outcomes and failure rates of OCA for patellofemoral joint chondral defects was performed with the Cochrane Database of Systematic Reviews, the Cochrane Central Register of Controlled Trials, PubMed, and MEDLINE from studies published between 1990 and 2017. Inclusion criteria were as follows: clinical outcomes and failure rates of OCA for the treatment of chondral defects in the patellofemoral joint, English language, minimum follow-up of 18 months, minimum study size of 5 patients, and human studies. The methodological quality of each study was assessed with a modified version of the Coleman methodology score. The systematic search identified 8 studies with a total of 129 patients. The methods of graft procurement and storage time included fresh (121 patients, 93.8%), and cryopreserved (8 patients, 6.2%) grafts. The mean survival rate was 87.9% at 5 years and 77.2% at 10 years. The following outcome scores showed significant improvement from pre- to postoperative status: modified d'Aubigné-Postel, International Knee Documentation Committee, Knee Society Score-Function, and Lysholm Knee Score. OCA of the patellofemoral joint results in improved patient-reported outcome measures with high patient satisfaction rates. Five- and 10-year survival rates of 87.9% and 77.2%, respectively, can be expected after this procedure. These findings should be taken with caution, as a high percentage of patellofemoral osteochondral allografts were associated with concomitant procedures; therefore, further research is warranted to determine the effect of isolated osteochondral transplantations.


Impaired Vascular Redox Signaling in the Vascular Complications of Obesity and Diabetes Mellitus.
Significance: Oxidative stress, a crucial regulator of vascular disease pathogenesis, may be involved in the vascular complications of obesity, systemic insulin resistance (IR), and diabetes mellitus (DM). Recent Advances: Excessive production of reactive oxygen species in the vascular wall has been linked with vascular disease pathogenesis. Recent evidence has revealed that vascular redox state is dysregulated in cases of obesity, systemic IR, and DM, potentially participating in the well-known vascular complications of these disease entities. Critical Issues: The detrimental effects of obesity and the metabolic syndrome on vascular biology have been extensively described at a clinical level. Further, vascular oxidative stress has often been associated with the presence of obesity and IR as well as with a variety of detrimental vascular phenotypes. However, the mechanisms of vascular redox state regulation under conditions of obesity and systemic IR, as well as their clinical relevance, are not adequately explored. In addition, the notion of vascular IR, and its relationship with systemic parameters of obesity and systemic IR, is not fully understood. In this review, we present all the important components of vascular redox state and the evidence linking oxidative stress with obesity and IR. Future Directions: Future studies are required to describe the cellular effects and the translational potential of vascular redox state in the context of vascular disease. In addition, further elucidation of the direct vascular effects of obesity and IR is required for better management of the vascular complications of DM.


Cognitive niches: an ecological model of strategy selection.
How do people select among different strategies to accomplish a given task? Across disciplines, the strategy selection problem represents a major challenge. We propose a quantitative model that predicts how selection emerges through the interplay among strategies, cognitive capacities, and the environment. This interplay carves out for each strategy a cognitive niche, that is, a limited number of situations in which the strategy can be applied, simplifying strategy selection. To illustrate our proposal, we consider selection in the context of 2 theories: the simple heuristics framework and the ACT-R (adaptive control of thought-rational) architecture of cognition. From the heuristics framework, we adopt the thesis that people make decisions by selecting from a repertoire of simple decision strategies that exploit regularities in the environment and draw on cognitive capacities, such as memory and time perception. ACT-R provides a quantitative theory of how these capacities adapt to the environment. In 14 simulations and 10 experiments, we consider the choice between strategies that operate on the accessibility of memories and those that depend on elaborate knowledge about the world. Based on Internet statistics, our model quantitatively predicts people's familiarity with and knowledge of real-world objects, the distributional characteristics of the associated speed of memory retrieval, and the cognitive niches of classic decision strategies, including those of the fluency, recognition, integration, lexicographic, and sequential-sampling heuristics. In doing so, the model specifies when people will be able to apply different strategies and how accurate, fast, and effortless people's decisions will be.


Antibodies to Porphyromonas gingivalis Indicate Interaction Between Oral Infection, Smoking, and Risk Genes in Rheumatoid Arthritis Etiology.
To investigate the role of the periodontal pathogen Porphyromonas gingivalis in the etiology of rheumatoid arthritis (RA) by analyzing the antibody response to the P gingivalis virulence factor arginine gingipain type B (RgpB) in relation to anti-citrullinated protein antibodies (ACPAs), smoking, and HLA-DRB1 shared epitope (SE) alleles in patients with periodontitis, patients with RA, and controls. Anti-RgpB IgG was measured by enzyme-linked immunosorbent assay in 65 periodontitis patients and 59 controls without periodontitis, and in 1,974 RA patients and 377 controls without RA from the Swedish population-based case-control Epidemiological Investigation of Rheumatoid Arthritis (EIRA) study. Autoantibody status, smoking habits, and genetic data were retrieved from the EIRA database. Differences in antibody levels were examined using the Mann-Whitney U test. Unconditional logistic regression was used to calculate odds ratios (ORs) with 95% confidence intervals (95% CIs) for the association of anti-RgpB IgG with different subsets of RA patients. Anti-RgpB antibody levels were significantly elevated in periodontitis patients compared to controls without periodontitis, in RA patients compared to controls without RA, and in ACPA-positive RA patients compared to ACPA-negative RA patients. There was a significant association between anti-RgpB IgG and RA (OR 2.96 [95% CI 2.00, 4.37]), which was even stronger than the association between smoking and RA (OR 1.37 [95% CI 1.07, 1.74]), and in ACPA-positive RA there were interactions between anti-RgpB antibodies and both smoking and the HLA-DRB1 SE. Our study suggests that the previously reported link between periodontitis and RA could be accounted for by P gingivalis infection, and we conclude that P gingivalis is a credible candidate for triggering and/or driving autoimmunity and autoimmune disease in a subset of RA patients.


Eruptive Milia Within a Tattoo: A Case Report and Review of the Literature.

Of the many tattoo reactions the most common are allergic, granulomatous, lichenoid, photosensitive, pseudolymphomatous, and infectious. Eruptive milia are a rare complication with only three prior reports in the English literature. A 19-year-old African American female presented with tiny, white papules confined within the margins of a tattoo. She denied trauma or associated symptoms at the site. Biopsy demonstrated deposits of black granular material within the dermis and a small infundibular cyst; a diagnosis of eruptive milia within tattoo was made. The milia responded to treatment with urea 40% cream and tretinoin 0.1% cream. Given its rarity, it is important to recognize the presentation of this disorder as other tattoo reactions require more aggressive management. While further research is necessary to determine the exact pathogenesis of this condition, the authors propose a mechanism along with a review of the literature to discuss management.

J Drugs Dermatol. 2017;16(6):621-624.
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Effect of body position on intraocular pressure in dogs without glaucoma.
To determine the effects of body position on intraocular pressure (IOP) in dogs without glaucoma. 24 healthy dogs with no evidence of glaucoma. Dogs underwent ophthalmic examinations to ensure that no IOP-affecting ocular diseases were present. Each dog was sequentially placed in dorsal recumbency, sternal recumbency, and sitting position. For each of the 3 positions, IOP in the right eye was measured by use of an applanation tonometer immediately after positioning (0 minutes) and after 3 and 5 minutes had elapsed. The initial body position was randomly assigned; each position followed the other positions an equal number of times, and IOP measurements were initiated immediately after moving from one body position to the next. Proparacaine hydrochloride (0.5%) was applied to the right eye immediately prior to IOP measurements. Intraocular pressure was affected by body position. During the 5-minute examination, IOP decreased significantly in dogs that were dorsally recumbent or sitting but did not change significantly in dogs that were sternally recumbent. For the 3 positions, overall mean IOP differed significantly at each time point (0, 3, and 5 minutes). Mean IOP in dorsal recumbency was significantly higher than that in sternal recumbency at 0 and at 3 minutes; although the former was also higher than that in sitting position at 3 minutes, that difference was not significant. Body position affects IOP in dogs. When IOP is measured in dogs, body position should be recorded and consistent among repeat evaluations.


The contribution of attentional lapses to individual differences in visual working memory capacity.
Attentional control and working memory capacity are important cognitive abilities that substantially vary between individuals. Although much is known about how attentional control and working memory capacity relate to each other and to constructs like fluid intelligence, little is known about how trial-by-trial fluctuations in attentional engagement impact trial-by-trial working memory performance. Here, we employ a novel whole-report memory task that allowed us to distinguish between varying levels of attentional engagement in humans performing a working memory task. By characterizing low-performance trials, we can distinguish between models in which working memory performance failures are caused by either (1) complete lapses of attention or (2) variations in attentional control. We found that performance failures increase with set-size and strongly predict working memory capacity. Performance variability was best modeled by an attentional control model of attention, not a lapse model. We examined neural signatures of performance failures by measuring EEG activity while participants performed the whole-report task. The number of items correctly recalled in the memory task was predicted by frontal theta power, with decreased frontal theta power associated with poor performance on the task. In addition, we found that poor performance was not explained by failures of sensory encoding; the P1/N1 response and ocular artifact rates were equivalent for high- and low-performance trials. In all, we propose that attentional lapses alone cannot explain individual differences in working memory performance. Instead, we find that graded fluctuations in attentional control better explain the trial-by-trial differences in working memory that we observe.


Elucidation of Conduction Mechanism in Graphene Nanoplatelets (GNPs)/Cement Composite Using Dielectric Spectroscopy.
Understanding the mechanisms that govern the conductive properties of multifunctional cement-materials is fundamental for the development of the new applications proposed to enhance the energy efficiency, safety and structural properties of smart buildings and infrastructures. Many fillers have been suggested to increase the electrical conduction in concretes; however, the processes involved are still not entirely known. In the present work, we investigated the effect of graphene nanoplatelets (1 wt% on the electrical properties of cement composites (OPC/GNPs). We found a decrease of the bulk resistivity in the composite associated to the enhancement of the charge transport properties in the sample. Moreover, the study of the dielectric properties suggests that the main contribution to conduction is given by water diffusion through the porous network resulting in ion conductivity. Finally, the results support that the increase of direct current in OPC/GNPs is due to pore refinement induced by graphene nanoplatelets.


Sterol regulatory element binding protein-dependent regulation of lipid synthesis supports cell survival and tumor growth.
Regulation of lipid metabolism via activation of sterol regulatory element binding proteins (SREBPs) has emerged as an important function of the Akt/mTORC1 signaling axis. Although the contribution of dysregulated Akt/mTORC1 signaling to cancer has been investigated extensively and altered lipid metabolism is observed in many tumors, the exact role of SREBPs in the control of biosynthetic processes required for Akt-dependent cell growth and their contribution to tumorigenesis remains unclear. We first investigated the effects of loss of SREBP function in non-transformed cells. Combined ablation of SREBP1 and SREBP2 by siRNA-mediated gene silencing or chemical inhibition of SREBP activation induced endoplasmic reticulum (ER)-stress and engaged the unfolded protein response (UPR) pathway, specifically under lipoprotein-deplete conditions in human retinal pigment epithelial cells. Induction of ER-stress led to inhibition of protein synthesis through increased phosphorylation of eIF2α. This demonstrates for the first time the importance of SREBP in the coordination of lipid and protein biosynthesis, two processes that are essential for cell growth and proliferation. SREBP ablation caused major changes in lipid composition characterized by a loss of mono- and poly-unsaturated lipids and induced accumulation of reactive oxygen species (ROS) and apoptosis. Alterations in lipid composition and increased ROS levels, rather than overall changes to lipid synthesis rate, were required for ER-stress induction.Next, we analyzed the effect of SREBP ablation in a panel of cancer cell lines. Importantly, induction of apoptosis following SREBP depletion was restricted to lipoprotein-deplete conditions. U87 glioblastoma cells were highly susceptible to silencing of either SREBP isoform, and apoptosis induced by SREBP1 depletion in these cells was rescued by antioxidants or by restoring the levels of mono-unsaturated fatty acids. Moreover, silencing of SREBP1 induced ER-stress in U87 cells in lipoprotein-deplete conditions and prevented tumor growth in a xenograft model. Taken together, these results demonstrate that regulation of lipid composition by SREBP is essential to maintain the balance between protein and lipid biosynthesis downstream of Akt and to prevent resultant ER-stress and cell death. Regulation of lipid metabolism by the Akt/mTORC1 signaling axis is required for the growth and survival of cancer cells.


Cytomegalovirus antibodies in dried blood spots: a minimally invasive method for assessing stress, immune function, and aging.
Cytomegalovirus (CMV) is a prevalent herpesvirus with links to both stress and aging. This paper describes and validates a minimally invasive method for assessing antibodies against CMV in finger stick whole blood spot samples for use as an indirect marker of an aspect of cell-mediated immunity. Analysis of CMV in dried blood spot samples (DBS) was based on modifications of a commercially available protocol for quantifying CMV antibodies in serum or plasma. The method was evaluated through analysis of precision, reliability, linearity, and correlation between matched serum and DBS samples collected from 75 volunteers. Correlation between DBS and plasma values was linear and high (Pearson correlation R = .96), and precision, reliability, and linearity of the DBS assay were within acceptable ranges. The validity of a DBS assay for CMV antibodies will enable its inclusion in population-based surveys and other studies collecting DBS samples in non-clinical settings, increasing scientific understanding of the interaction of social and biological stress and immune function.


A baseline study on the importance of bovines for human Schistosoma japonicum infection around Poyang Lake, China.
We hypothesize that bovine infections are responsible for the persistence of human schistosomiasis transmission in the Yangtze marshlands of China. To test this hypothesis, we are carrying out a comparative intervention among four administrative villages in the Poyang Lake region, Jiangxi Province, two of which are experimental and two are control. The primary design involves treating, at the onset of the study, all the inhabitants in all four villages with praziquantel and all the bovines in two villages (the experimental or intervention villages). Following treatment, rates of reinfection in people of all villages, and in bovines in the experimental villages, will be assessed as will the ongoing prevalence of infection in bovines in the control villages. Before treatment, the prevalence and intensity of infection among humans and bovines was ascertained in the four villages. Our study design and baseline information are presented here, along with a description of the ecology of the study villages.


Prognosis of myelodysplastic syndromes.
The myelodysplastic syndromes (MDS) are a very complex group of hematopoietic disorders. The degree of complexity relates not only to the intrinsic pathobiological characteristics of the disease, but also to the group of patients whom it affects most frequently: older individuals or those who have been exposed to prior forms of chemotherapy. It is therefore crucial to develop clinical tools to predict with a certain degree of precision the prognosis and outcome for patients with specific subtypes of MDS in specific clinical situations. At the present time, patients with MDS are diagnosed using a set of well-established histopathological criteria. Prognosis is established using classifications that include morphological features, percentage of blasts, and clinical and molecular characteristics such as peripheral cytopenias and cytogenetics. The International Prognostic Scoring System (IPSS) is a classic example of this type of classification. Over the last 5 years, there has been an intense effort to develop new prognostic systems for MDS, and new molecular alterations with potential prognostic value have been discovered. Over the same period of time, several new therapeutic interventions have been developed for patients with MDS. Biomarkers of response to these agents, in particular for the hypomethylating agents, are needed to predict clinical benefit. This review summarizes current prognostic models of MDS and new molecular alterations with potential prognostic potential.


Understanding the burden of bacterial sexually transmitted infections and Trichomonas vaginalis among black Caribbeans in the United Kingdom: Findings from a systematic review.
In the UK, people of black Caribbean (BC) ethnicity continue to be disproportionately affected by bacterial sexually transmitted infections (STIs) and Trichomonas vaginalis (TV). We systematically reviewed evidence on the association between bacterial STIs/TV and ethnicity (BC compared to white/white British (WB)) accounting for other risk factors; and differences between these two ethnic groups in the prevalence of risk factors associated with these STIs, sexual healthcare seeking behaviours, and contextual factors influencing STI risk. Studies presenting relevant evidence for participants aged ≥14 years and living in the UK were eligible for inclusion. A pre-defined search strategy informed by the inclusion criteria was developed. Eleven electronic databases were searched from the start date to September-October 2016. Two researchers independently screened articles, extracted data using a standardised proforma and resolved discrepancies in discussion with a third researcher. Descriptive summaries of evidence are presented. Meta-analyses were not conducted due to variation in study designs. Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines were followed. Of 3815 abstracts identified, 15 articles reporting quantitative data were eligible and included in the review. No qualitative studies examining contextual drivers of STI risk among people of BC ethnicity were identified. Compared to the white/WB ethnic group, the greater STI/TV risk among BCs was partially explained by variations in socio-demographic factors, sexual behaviours, and recreational drug use. The prevalence of reporting early sexual debut (<16 years), concurrency, and multiple partners was higher among BC men compared to white/WB men; however, no such differences were observed for women. People of BC ethnicity were more likely to access sexual health services than those of white/WB ethnicity. Further research is needed to explore other drivers of the sustained higher STI/TV prevalence among people of BC ethnicity. Developing holistic, tailored interventions that address STI risk and target people of BC ethnicity, especially men, could enhance STI prevention.


Relationship between cell volume and ion transport in the early distal tubule of the Amphiuma kidney.
The roles of apical and basolateral transport mechanisms in the regulation of cell volume and the hydraulic water permeabilities (Lp) of the individual cell membranes of the Amphiuma early distal tubule (diluting segment) were evaluated using video and optical techniques as well as conventional and Cl-sensitive microelectrodes. The Lp of the apical cell membrane calculated per square centimeter of tubule is less than 3% that of the basolateral cell membrane. Calculated per square centimeter of membrane, the Lp of the apical cell membrane is less than 40% that of the basolateral cell membrane. Thus, two factors are responsible for the asymmetry in the Lp of the early distal tubule: an intrinsic difference in the Lp per square centimeter of membrane area, and a difference in the surface areas of the apical and basolateral cell membranes. Early distal tubule cells do not regulate volume after a reduction in bath osmolality. This cell swelling occurs without a change in the intracellular Cl content or the basolateral cell membrane potential. In contrast, reducing the osmolality of the basolateral solution in the presence of luminal furosemide diminishes the magnitude of the increase in cell volume to a value below that predicted from the change in osmolality. This osmotic swelling is associated with a reduction in the intracellular Cl content. Hence, early distal tubule cells can lose solute in response to osmotic swelling, but only after the apical Na/K/Cl transporter is blocked. Inhibition of basolateral Na/K ATPase with ouabain results in severe cell swelling. This swelling in response to ouabain can be inhibited by the prior application of furosemide, which suggests that the swelling is due to the continued entry of solutes, primarily through the apical cotransport pathway.


Hepatitis B virus transactivator X protein is not tumorigenic in transgenic mice.
The hepatitis B virus X protein acts as a transcriptional transactivator in vitro. To elucidate possible biological effects of X protein on liver cells in vivo, we generated four lines of transgenic mice carrying the X gene open reading frame under the control of the human alpha-1-antitrypsin regulatory region. The plasmid construct used to introduce the transgene was shown to encode a 16-kDa X protein with transactivating capability. The expression of X protein was detectable in liver tissue of transgenic animals of three of the lines by immunoblot analysis; levels of expression were highest in the first month after birth of the animals. Over 80 animals from the expressing lines were examined histologically. Most transgenic mice, some of which were observed for up to 2 years, remained normal. However, a few transgenic animals developed mild focal hepatitis, nuclear pleomorphism, focal necrosis, and/or nodular hyperplasia in the liver. Increased mitotic activity of hepatocytes also was observed. We conclude that, at the level of expression achieved in these transgenic mice, the hepatitis B virus transcriptional transactivator X protein alone does not appear to mediate the development of serious liver damage or hepatocellular carcinomas.


Isolation and characterization of multiple forms of rat liver UDP-glucuronate glucuronosyltransferase.
UDP-glucuronosyltransferase (EC 188.8.131.52) activity was solubilized from male Wistar rat liver microsomal fraction in Emulgen 911, and six fractions with the transferase activity were separated by chromatofocusing on PBE 94 (pH 9.4 to 6.0). Fraction I was further separated into Isoforms Ia, Ib and Ic by affinity chromatography on UDP-hexanolamine-Sepharose 4B. UDP-glucuronosyltransferase in Fraction III was further purified by rechromatofocusing (pH 8.7 to 7.5). UDP-glucuronosyltransferases in Fractions IV and V were purified by UDP-hexanolamine-Sepharose chromatography. The transferase isoforms in Fractions II, III, IV and V were finally purified by h.p.l.c. on a TSK G 3000 SW column. Purified UDP-glucuronosyltransferase Isoforms Ia (Mr 51,000), Ib (Mr 52,000), Ic (Mr 56,000), II (Mr 52,000), IV (Mr 53,000) and V (Mr 53,000) revealed single Coomassie Blue-stained bands on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Isoform III enzyme showed two bands of Mr 52,000 and 53,000. Comparison of the amino acid compositions by the method of Cornish-Bowden [(1980) Anal. Biochem. 105, 233-238] suggested that all UDP-glucuronosyltransferase isoforms are structurally related. Reverse-phase h.p.l.c. of tryptic peptides of individual isoforms revealed distinct 'maps', indicating differences in primary protein structure. The two bands of Isoform III revealed distinct electrophoretic peptide maps after limited enzymic proteolysis. After reconstitution with phosphatidylcholine liposomes, the purified isoforms exhibited distinct but overlapping substrate specificities. Isoform V was specific for bilirubin glucuronidation, which was not inhibited by other aglycone substrates. Each isoform, except Ia, was identified as a glycoprotein by periodic acid/Schiff staining.


U-Pb ages from the neoproterozoic Doushantuo Formation, China.
U-Pb zircon dates from volcanic ash beds within the Doushantuo Formation (China) indicate that its deposition occurred between 635 and 551 million years ago. The base records termination of the global-scale Marinoan glaciation and is coeval with similar dated rocks from Namibia, indicating synchronous deglaciation. Carbon isotopic and sequence-stratigraphic data imply that the spectacular animal fossils of the Doushantuo Formation are for the most part younger than 580 million years old. The uppermost Doushantuo Formation contains a pronounced negative carbonate carbon isotopic excursion, which we interpret as a global event at circa 551 million years ago.


Physiological and behavioral responses reveal 9-month-old infants' sensitivity to pleasant touch.
Caregiving touch has been shown to be essential for the growth and development of human infants. However, the physiological and behavioral mechanisms that underpin infants' sensitivity to pleasant touch are still poorly understood. In human adults, a subclass of unmyelinated peripheral nerve fibers has been shown to respond preferentially to medium-velocity soft brushing. It has been theorized that this privileged pathway for pleasant touch is used for close affiliative interactions with conspecific individuals, especially between caregivers and infants. To test whether human infants are sensitive to pleasant touch, we examined arousal (heart rate) and attentional engagement (gaze shifts and duration of looks) to varying velocities of brushing (slow, medium, and fast) in 9-month-old infants. Our results provide physiological and behavioral evidence that sensitivity to pleasant touch emerges early in development and therefore plays an important role in regulating human social interactions.