Adjuvant immunotherapy for locally advanced renal cell carcinoma.
Locally advanced renal cell carcinoma (RCC) presents a therapeutic challenge due to 20-40% relapse risk post-nephrectomy. There has been substantial interest in utilizing immunotherapy interrupting the PD-1/PD-L1 axis in the perioperative space, especially in the adjuvant setting, in order to minimize such risk. We conducted a PubMed search using the terms 'adjuvant' and 'RCC.' We begin by examining landmark studies in the postoperative space for locally advanced RCC, with special emphasis on immunotherapeutic biologics. Important considerations are outlined in an effort to explain the conflicting data on the benefit of adjuvant immunotherapy as well as to adequately assess the magnitude of potential benefit of the recently approved adjuvant pembrolizumab. Relevant contemporary challenges and opportunities as well as future directions of the field are also discussed. Systemic immunotherapy with monoclonal antibodies targeting the PD-1/PD-L1 axis likely holds promise, either alone or potentially in combinations, in minimizing recurrence risk for locally advanced RCC. However, emphasis on post-protocol care, robust endpoint selection, and continued work and validation on predictive biomarkers are needed to confidently select those patients that may benefit the most and minimize biologic and financial toxicity.


[Factors of ischemic lesions in the kidney and its resection].
Resection of kidney is an organ-sparing operation finding an increasingly wider application. It is designed to preserve the maximum amount of functionally competent renal parenchyma. However, surgery produces multiple effects on the kidney resulting in ischemic lesions and dysfunction which has serious consequences for the clinical course of the early post-operative period and long-term outcome of the treatment. The causes and after-effects of ischemia are considered, much attention is given to its pathogenetic mechanisms and the process of healing of renal parenchyma depending on the surgical modality and method of hemostasis.


Water at polar and nonpolar solid walls.
Recent progress in simulating the properties of interfacial water at hard hydrophobic and hydrophilic surfaces is reviewed and compared to results for the air/water interface. The authors discuss static properties such as the equilibrium contact angle, the depletion layer thickness, and the orientation of interfacial water molecules. Relations between these properties, e.g., the relation between the contact angle and the thickness of the depletion layer which is experimentally observed on hydrophobic surfaces, are emphasized. For a hydrophilic sapphire surface, the authors discuss the influence of geometry and density of polar surface groups on the interfacial water structure. They discuss nonequilibrium effects arising in laminar shear flows, where the classic no-slip hydrodynamic boundary condition is violated at hydrophobic interfaces. They discuss the arising slip and relate it to static properties of the solid hydrophobic/water interface.


Micronutrients and anaemia.
Micronutrient deficiencies and anaemia remain as major health concerns for children in Bangladesh. Among the micronutrient interventions, supplementation with vitamin A to children aged less than five years has been the most successful, especially after distribution of vitamin A was combined with National Immunization Days. Although salt sold in Bangladesh is intended to contain iodine, much of the salt does not contain iodine, and iodine deficiency continues to be common. Anaemia similarly is common among all population groups and has shown no sign of improvement even when iron-supplementation programmes have been attempted. It appears that many other causes contribute to anaemia in addition to iron deficiency. Zinc deficiency is a key micronutrient deficiency and is covered in a separate paper because of its importance among new child-health interventions.


In their own voices: rural African American youth speak out about community-based HIV prevention interventions.
The HIV epidemic is a major public health problem in the United States, particularly among rural African American adolescents and young adults. We sought to explore young, rural African American's perspectives about key programmatic components to consider when designing youth-targeted, community- based HIV prevention interventions. We report data from four focus groups with adolescents and young adults aged 16 to 24 (n = 38) conducted as part of a community-based participatory research (CBPR) project designed to develop multilevel HIV risk reduction interventions in two rural North Carolina communities with high HIV rates. Analysis was performed by academic and community partners using a modified grounded theory approach to content analysis. Interventions should target preadolescents and early adolescents rather than older adolescents and young adults in an effort to "catch them while they're young." Intervention developers should obtain input from local young people regarding critical programmatic components, such as whom to employ as study recruiters and intervention leaders; intervention format and delivery options, acceptable recruitment and intervention locations, and incentive structures. Participants believe selecting community collaborators representing varied community sectors is critical. Important barriers to address included limited transportation, discomfort communicating about sexual issues, lack of community interest in HIV prevention, and unwillingness to acknowledge and address sexual activity among adolescents. When designing HIV/AIDS prevention interventions, targeting young people, it is important to form academic-community partnerships that ensure young people's perspectives are integral to the intervention development process.


Regeneration of the heart in diabetes by selective copper chelation.
Heart disease is the major cause of death in diabetes, a disorder characterized by chronic hyperglycemia and cardiovascular complications. Although altered systemic regulation of transition metals in diabetes has been the subject of previous investigation, it is not known whether changed transition metal metabolism results in heart disease in common forms of diabetes and whether metal chelation can reverse the condition. We found that administration of the Cu-selective transition metal chelator trientine to rats with streptozotocin-induced diabetes caused increased urinary Cu excretion compared with matched controls. A Cu(II)-trientine complex was demonstrated in the urine of treated rats. In diabetic animals with established heart failure, we show here for the first time that 7 weeks of oral trientine therapy significantly alleviated heart failure without lowering blood glucose, substantially improved cardiomyocyte structure, and reversed elevations in left ventricular collagen and beta(1) integrin. Oral trientine treatment also caused elevated Cu excretion in humans with type 2 diabetes, in whom 6 months of treatment caused elevated left ventricular mass to decline significantly toward normal. These data implicate accumulation of elevated loosely bound Cu in the mechanism of cardiac damage in diabetes and support the use of selective Cu chelation in the treatment of this condition.


Synthesis, Photophysical and Computational Study of Novel Coumarin-based Organic Dyes.
A series of novel coumarin pyrazoline moieties combined with tetrazoles, 3-(1-phenyl-4-(1H-tetrazol-5-yl)-1H-pyrazol-3-yl)-2H-chromen-2-one, 6-chloro-3-(1-phenyl-4-(1H-tetrazol-5-yl)-1H-pyrazol-3-yl)-2H-chromen-2-one, 6-bromo-3-(1-phenyl-4-(1H-tetrazol-5-yl)-1H-pyrazol-3-yl)-2H-chromen-2-one and 6-bromo-3-(1-(4-bromophenyl)-4-(1H-tetrazol-5-yl)-1H pyrazol-3-yl)-2H-chromen-2-one7(a-d), were designed and synthesized. Single crystal X-ray diffraction and their interactions were studied by Hirshfeld surface analysis. Thermal stabilities and electrochemical properties of these compounds were examined from differential scanning calorimetry (DSC), thermogravimetric (TGA) and cyclic voltammetric (CV) studies. Their spectroscopic properties were analyzed in various alcohols and general solvents by UV-Vis absorption, fluorescence and time-resolved spectroscopy. In addition, the ground and excited state electronic properties were investigated using density functional theory (DFT). The calculated highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) and energy band gap (Eg ) values have revealed the effect of substitution of halogens. The substitution has equally affected the ground and excited states of 7(a-d) compounds. The solvatochromism on absorption, fluorescence spectra and fluorescence lifetimes of these compounds was investigated. All these results showed the chromen-2-one of pyrazoline tetrazole derivatives could play an important role in photonic and electronic devices.


Bayesian single-step genomic evaluations combining local and foreign information in Walloon Holsteins.
Most dairy cattle populations found in different countries around the world are small to medium sized and use many artificial insemination bulls imported from different foreign countries. The Walloon population in the southern part of Belgium is a good example for such a small-scale population. Wallonia has also a very active community of Holstein breeders requesting high level genetic evaluation services. Single-step Genomic BLUP (ssGBLUP) methods allow the simultaneous use of genomic, pedigree and phenotypic information and could reduce potential biases in the estimation of genomically enhanced breeding values (GEBV). Therefore, in the context of implementing a Walloon genomic evaluation system for Holsteins, it was considered as the best option. However, in contrast to multi-step genomic predictions, natively ssGBLUP will only use local phenotypic information and is unable to use directly important other sources of information coming from abroad, for example Multiple Across Country Evaluation (MACE) results as provided by the Interbull Center (Uppsala, Sweden). Therefore, we developed and implemented single-step Genomic Bayesian Prediction (ssGBayes), as an alternative method for the Walloon genomic evaluations. The ssGBayes method approximated the correct system of equations directly using estimated breeding values (EBV) and associated reliabilities (REL) without any explicit deregression step. In the Walloon genomic evaluation, local information refers to Walloon EBV and REL and foreign information refers to MACE EBV and associated REL. Combining simultaneously all available genotypes, pedigree, local and foreign information in an evaluation can be achieved but adding contributions to left-hand and right-hand sides subtracting double-counted contributions. Correct propagation of external information avoiding double counting of contributions due to relationships and due to records can be achieved. This ssGBayes method computed more accurate predictions for all types of animals. For example, for genotyped animals with low Walloon REL (<0.25) without MACE results but sired by genotyped bulls with MACE results, the average increase of REL for the studied traits was 0.38 points of which 0.08 points could be traced to the inclusion of MACE information. For other categories of genotyped animals, the contribution by MACE information was also high. The Walloon genomic evaluation system passed for the first time the Interbull GEBV tests for several traits in July 2013. Recent experiences reported here refer to its use in April 2016 for the routine genomic evaluations of milk production, udder health and type traits. Results showed that the proposed methodology should also be of interest for other, similar, populations.


Pyelonephritis Caused Solely by Escherichia hermanii.
In contrast with Escherichia coli, the association of E. hermanii with urinary tract infections has not been described. In this case, E. hermanii was the sole isolate recovered from urine specimens of a pyelonephritis patient. The organism was found to be susceptible to piperacillin-tazobactam, ceftazidime, cefazolin, cefixime, aztreonam, gentamicin, tobramycin, imipenem, meropenem and amikacin, and resistant to amoxicillin. Antibiotic treatment was initiated with oral cefixime (400 mg every 24 hours). The symptoms were relieved within 72 hours after therapy. A urine sample was taken seven days after antibiotic therapy. E. hermanii was no longer isolated. The present case demonstrates that the uropathogenic E. hermanii clone can cause destruction of the kidneys. During asymptomatic bacteriuria or cystitis, the bacteria remain in the urinary tract. Even when pyelonephritis develops, inflammatory response of the host is still restricted to the urinary tract. These signs mean that uropathogenic E. hermanii may be not very virulent.


Necrostatin-1 ameliorates the pathogenesis of experimental autoimmune encephalomyelitis by suppressing apoptosis and necroptosis of oligodendrocyte precursor cells.
Multiple sclerosis (MS) is an autoimmune disease of the central nervous system characterized by neuronal demyelination. MS pathogenesis occurs via multiple mechanisms, and is mediated in part by oligodendrocyte apoptosis and a robust inflammatory response. In the present study, Necrostatin-1 (Nec-1), a specific inhibitor of the receptor-interacting protein 1 kinase domain, was revealed to effectively alleviate the severity and pathological damage associated with experimental autoimmune encephalomyelitis (EAE), a commonly used mouse model of MS. In addition, treatment with Nec-1 significantly decreased the number of lesions and inflammatory cell infiltrates in spinal cord tissues, as well as the production of associated pro-inflammatory cytokines, including tumor necrosis factor α (TNFα), interferon-γ and interleukin-1β. Nec-1 also suppressed TNFα + zVAD-fmk-induced apoptosis and necroptosis in primary oligodendrocyte precursor cells. The present study revealed that Nec-1 effectively attenuated the progression of EAE by suppressing apoptosis and necroptosis in oligodendrocytes, and represents a potential novel therapeutic agent for the treatment of MS.


Outcome in African Americans and other minorities in the Sudden Cardiac Death in Heart Failure Trial (SCD-HeFT).
The SCD-HeFT demonstrated that implantable cardioverter/defibrillator (ICD) therapy significantly improved survival compared to medical therapy alone in stable moderately symptomatic heart failure patients with an ejection fraction < or = 35%. The purpose of this report is to describe the outcomes in African Americans (AAs) and other minorities. Of 2521 patients enrolled, 23% were minorities and 17% were AAs. Baseline demographic, clinical variables, socioeconomic status, and long-term outcomes were compared according to race. Two major prespecified subgroups were examined: heart failure cause (ischemic vs nonischemic) and New York Heart Association class (II vs III). At baseline, compared to whites, AAs were younger and had more nonischemic heart failure, lower ejection fractions, worse New York Heart Association functional class, and higher prevalence of a history of nonsustained ventricular tachycardia. Comparable percentages of whites and AAs held paid jobs, but whites had a significantly higher educational level and household income (P = .001). Compliance with ICD implantation and medical therapy was comparable in both subgroups. No significant difference was observed in the rate of ICD discharge among whites and AAs. Adjusted mortality risk was significantly higher in AAs compared to whites (hazard ratio 1.27, P = .038). Mortality was equally reduced in both race groups receiving ICD therapy compared to placebo (hazard ratio 0.65 in AAs and 0.73 in whites). Survival benefits from ICD therapy in SCD-HeFT were not dependent on race. In addition, in this clinical trial setting, there was no evidence that AAs were less willing to accept ICD therapy than whites.


Switching from omalizumab to mepolizumab: real-life experience from Southern Italy.
Current availability of several biologic treatments for severe asthma makes it possible to choose the most appropriate for each patient. Sometimes a good percentage of patients with severe asthma may be eligible for biologics that target either the allergic phenotype or the eosinophilic one, but not all respond to that selected as first choice. The aim of our real-life study was to assess whether, for patients with severe eosinophilic allergic asthma, not previously controlled by the anti-IgE omalizumab, the shift to another biologic targeting interleukin-5, such as mepolizumab, may represent a good therapeutic choice. A total of 41 consecutive patients with severe, persistent allergic, eosinophilic asthma, uncontrolled despite treatment with omalizumab, were enrolled in seven certified Clinical Respiratory Units of Southern Italy (Catania, Catanzaro, Foggia, Bari, Palermo, and two University Respiratory Units of Naples) and shifted to mepolizumab without a wash-out period. Data at baseline, after at least 12 months of therapy with omalizumab, and after at least 12 months of treatment with mepolizumab were collected. After at least 12 months of therapy with mepolizumab, patients experienced a significant decrease in the number of exacerbations/year (5.8 ± 1.8 versus 0.7 ± 0.9, p < 0.0001), an increment of asthma control test score (12 ± 2.7 versus 21.9 ± 2.7, p < 0.0001), an increase in pre-bronchodilator forced expiratory volume in 1 s (1.56 ± 0.45 l versus 1.86 ± 0.52 l, p < 0.0001), and a reduction of blood eosinophils (584 ± 196 cells/µl versus 82 ± 56 cells/µl, p < 0.0001). The percentage of patients who were dependent on corticosteroids significantly decreased from 46% at baseline to 5% during treatment with mepolizumab. Results of our real-life multicentric experience confirms that the shift to mepolizumab could be a good therapeutic strategy in severe eosinophilic allergic asthma not previously controlled by omalizumab. The reviews of this paper are available via the supplemental material section.


[New principles for obtaining protective antigens from deep cutlures of enterobacteria].
A model of S. typhi submerged culture was used; application of the principle of sparing extraction of surface protective antigens by the action of sublethal concentrations of two damaging agents differing by the mechanism of inhibition of the processes of vital activity of the microorganisms was founded. The principle is based on regularities characteristic of cells of bacterial pupulations: a) nonhomogeneity of individual cells of the population by the resistance to the damaging agents, and b) independence of distribution in the population cells of resistance to different damaging agents. The following typhoid antigens were obtained by combined action of sublethal concentrations of hydrogen peroxide (0.25%) and formalin (0.1%): antigens with a full antigenic structure (inducing formation of O-, Vi- and H-antibodies), of low toxicity (V50 - 0.65+/-19 mg) and highly immunogenic (AU50- 0.45+/-0.25 mug) in experiments on mice.


A Close-up Look at the Chemical Space of Commercially Available Building Blocks for Medicinal Chemistry.
The ability to efficiently synthesize desired compounds can be a limiting factor for chemical space exploration in drug discovery. This ability is conditioned not only by the existence of well-studied synthetic protocols but also by the availability of corresponding reagents, so-called building blocks (BBs). In this work, we present a detailed analysis of the chemical space of 400 000 purchasable BBs. The chemical space was defined by corresponding synthons─fragments contributed to the final molecules upon reaction. They allow an analysis of BB physicochemical properties and diversity, unbiased by the leaving and protective groups in actual reagents. The main classes of BBs were analyzed in terms of their availability, rule-of-two-defined quality, and diversity. Available BBs were eventually compared to a reference set of biologically relevant synthons derived from ChEMBL fragmentation, in order to illustrate how well they cover the actual medicinal chemistry needs. This was performed on a newly constructed universal generative topographic map of synthon chemical space that enables visualization of both libraries and analysis of their overlapped and library-specific regions.


Differential expression of IGF family members in heat-stressed embryos produced in vitro from OPU-derived oocytes of Nelore (Bos indicus) and Holstein (Bos taurus) cows.
The IGF system is related to embryo quality. We aim to determine the effect of the heat stress on the mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4, and PAPPA in in vitro production (IVP) blastocysts from Nelore and Holstein after ovum pick up (OPU) to better understand the differences between these breeds. Oocytes from four Nelore and seven Holstein were collected in six OPU sessions. Following in vitro maturation and fertilization using six Nelore or Holstein sires, embryos were divided into control (cultured at 39°C) and heat stress (HS; exposed to 41°C for 9 h). Blastocysts were submitted to RNA extraction. The IGF1 expression was higher in blastocysts under HS in both breeds, and the expression of IGFBP2 and IGFBP4 was higher in Holstein blastocysts under HS. The high PAPPA expression and the low expression of IGFBP2 and IGFBP4 are associated with a more efficient degradation of IGFBPs, which results in greater IGF bioavailability in Nelore blastocysts and may contribute to the superior HS tolerance in Nelore, when compared to Holstein.


Detection of alpha 2-macroglobulin, alpha 1-protease inhibitor, and neutral protease-antiprotease complexes within liver granulomas of Schistosoma mansoni-infected mice.
In schistosomiasis mansoni the parasite egg-induced granulomatous tissue inflammations resolve by fibrosis. Intralesional collagen synthesis and deposition are influenced by collagenase, elastase activity that is diminished at the chronic stage of the disease. To determine the cause of diminished neutral protease activity, the authors determined levels of the antiprotease/alpha 2-macroglobulin (alpha 2M) and alpha 1-protease inhibitor (alpha 1Pi) in extracts or secretions of liver granulomas of infected mice. By ELISA, both antiproteases were detected in granuloma-derived substances, as well as supernatants of cultured, adherent granuloma macrophages. In all samples, alpha 2M was the predominant inhibitor. Antiprotease levels were similar in granuloma-derived samples obtained from acutely and chronically infected mice. However, supernatants of cultured adherent macrophages isolated from granulomas of mice with acute infection contained levels of protease inhibitors several times higher than those of similar preparations obtained from chronically infected animals. Gel filtration of samples on Sephacryl S-200 columns did not separate collagenase and elastase from protease inhibitors. By chromatofocusing, a few inhibitor-free collagenase as well as enzyme-free alpha 2M and alpha 1Pi-active peaks were eluted. The bulk of the material that eluted at the acidic region contained protease-antiprotease activity indicating the presence of enzyme-inhibitor complexes. The intragranulomatous presence of antiproteases complexed with protease enzymes emphasizes their importance in the possible enhancement of fibrosis.


Optimal portfolio design to reduce climate-related conservation uncertainty in the Prairie Pothole Region.
Climate change is likely to alter the spatial distributions of species and habitat types but the nature of such change is uncertain. Thus, climate change makes it difficult to implement standard conservation planning paradigms. Previous work has suggested some approaches to cope with such uncertainty but has not harnessed all of the benefits of risk diversification. We adapt Modern Portfolio Theory (MPT) to optimal spatial targeting of conservation activity, using wetland habitat conservation in the Prairie Pothole Region (PPR) as an example. This approach finds the allocations of conservation activity among subregions of the planning area that maximize the expected conservation returns for a given level of uncertainty or minimize uncertainty for a given expected level of returns. We find that using MPT instead of simple diversification in the PPR can achieve a value of the conservation objective per dollar spent that is 15% higher for the same level of risk. MPT-based portfolios can also have 21% less uncertainty over benefits or 6% greater expected benefits than the current portfolio of PPR conservation. Total benefits from conservation investment are higher if returns are defined in terms of benefit-cost ratios rather than benefits alone. MPT-guided diversification can work to reduce the climate-change-induced uncertainty of future ecosystem-service benefits from many land policy and investment initiatives, especially when outcomes are negatively correlated between subregions of a planning area.


A Multitrait-Multimethod Matrix Investigation of Hoarding.
Hoarding is a serious and potentially life-threatening mental health problem that, until recently, was considered a subtype of OCD. However, recent research suggests it is distinct and more prevalent than OCD. Three key defining features have emerged in factor analytic studies of hoarding scales: excessive acquisition, difficulty discarding, and excessive clutter. Covariation among these defining features has received limited attention. The primary aim of the current study was to examine the role of the three key features in defining hoarding disorder. Convergent and discriminant validity of the three hoarding factors were examined in a multitrait-multimethod matrix. A secondary aim was to examine the extent to which each hoarding feature distinguished individuals meeting criteria for hoarding from those with OCD and community controls. Although the three-factor model provided an adequate fit for the data and convergent validities were high, the hoarding factors evidenced poor discriminant validity across measures. The findings provide preliminary support for a more parsimonious merging of the clutter, acquisition, and discarding subscales versus parsing out subscale scores. Specifically, the active acquisition of items, buildup of clutter, and difficulty discarding accumulated possessions co-occurred strongly enough to be considered a unidimensional construct. Thus, these symptoms were less attributable to separate phenomena and better conceived as part of a cohesive hoarding phenotype. Each of the three factors discriminated hoarding participants from OCD patients and community controls, but did not discriminate the latter two groups. The findings have implications for treating acquisition as a specifier in DSM-5.


A stochastic model for colloid transport and deposition.
Profiles of retained colloids in porous media have frequently been observed to be hyper-exponential or non-monotonic with transport depth under unfavorable attachment conditions, whereas filtration theory predicts an exponential profile. In this work we present a stochastic model for colloid transport and deposition that allows various hypotheses for such deviations to be tested. The model is based on the conventional advective dispersion equation that accounts for first-order kinetic deposition and release of colloids. One or two stochastic parameters can be considered in this model, including the deposition coefficient, the release coefficient, and the average pore water velocity. In the case of one stochastic parameter, the probability density function (PDF) is characterized using log-normal, bimodal log-normal, or a simple two species/region formulation. When two stochastic parameters are considered, then a joint log-normal PDF is employed. Simulation results indicated that variations in the deposition coefficient and the average pore water velocity can both produce hyper-exponential deposition profiles. Bimodal formulations for the PDF were also able to produce hyper-exponential profiles, but with much lower variances in the deposition coefficient. The shape of the deposition profile was found to be very sensitive to the correlation of deposition and release coefficients, and to the correlation of pore water velocity and deposition coefficient. Application of the developed stochastic model to a particular set of colloid transport and deposition data indicated that chemical heterogeneity of the colloid population could not fully explain the observed behavior. Alternative interpretations were therefore proposed based on variability of the pore size and the water velocity distributions.


Upregulation of P-glycoprotein in rat hepatoma rho(o) cells: implications for drug-DNA interactions.
Rat hepatoma cells lacking mitochondrial DNA (rho(o) cells) were used as a model system to examine the possible roles of mitochondrial DNA as a target for the DNA-acting anticancer drug Adriamycin (doxorubicin). The rho(o) cells were 45-fold less sensitive to Adriamycin than the parental rho+ cells containing mitochondrial DNA. Other non-DNA-acting drugs also exhibited similar behaviour, and this was shown to be due to a multidrug resistance (MDR) phenotype in the rho(o) cells. This was indicated by confocal microscopy where rho+ cells exhibited thirteenfold higher cellular levels of Adriamycin than rho(o) cells. Upregulation (tenfold) of P-glycoprotein in rho(o) cells was also confirmed by Northern dot blot analysis. Since the MDR phenotype is present in rho(o) cells and upregulation of P-glycoprotein is maintained in these cells, rho(o) cells are not a good model system for drug-DNA studies (where the drug is susceptible to extrusion by P-glycoprotein), and any such results obtained with this system must be treated with considerable caution.


Sandwich fusion of CBM9_2 to enhance xylanase thermostability and activity.
Used as model for sandwich fusion, a mesophilic Aspergillus niger GH11 xylanase (Xyn) was fused into C2-Xyn-C2 with a thermophilic Thermotaga maritima GH10 xylanase carbohydrate-binding module CBM9_2 (C2). Linearized plasmids C2-pET20b-C2-Xyn were amplified from template pET20b-Xyn-C2 with a 4.3 kb C2-pET20b megaprimer, ligated into circular plasmids in blunt-end ligation, and transformed into E. coli BL21 (DE3) cells. The C2-Xyn-C2 had optimum activity at 45 °C and pH 4.2, a 2.85 h thermal inactivation half-life at 80 °C and a 8.69 h at 50 °C, with the 8.69 h value 24.8-, 7.5-, and 7.1-fold longer than the Xyn and single terminal fusion enzymes Xyn-C2, and C2-Xyn. Thermodynamics showed that the enzyme had a 1.8 °C higher melting temperature, lower values ΔS, ΔΔG, and a denser structure than the Xyn. Kinetics showed that the C2-Xyn-C2 catalytic efficiency was 1.2-~6-fold and 2.7-~7.9-fold higher on beechwood and oat-spelt xylan than those of the enzymes Xyn, Xyn-C2, and C2-Xyn. The sandwich fusion evolved the xylanase with "armor-hands" to enhance simultaneously thermostability and activity in quality.


Monolayer Support Control and Precise Colloidal Nanocrystals Demonstrate Metal-Support Interactions in Heterogeneous Catalysts.
Electronic and geometric interactions between active and support phases are critical in determining the activity of heterogeneous catalysts, but metal-support interactions are challenging to study. Here, it is demonstrated how the combination of the monolayer-controlled formation using atomic layer deposition (ALD) and colloidal nanocrystal synthesis methods leads to catalysts with sub-nanometer precision of active and support phases, thus allowing for the study of the metal-support interactions in detail. The use of this approach in developing a fundamental understanding of support effects in Pd-catalyzed methane combustion is demonstrated. Uniform Pd nanocrystals are deposited onto Al2 O3 /SiO2 spherical supports prepared with control over morphology and Al2 O3 layer thicknesses ranging from sub-monolayer to a ≈4 nm thick uniform coating. Dramatic changes in catalytic activity depending on the coverage and structure of Al2 O3 situated at the Pd/Al2 O3 interface are observed, with even a single monolayer of alumina contributing an order of magnitude increase in reaction rate. By building the Pd/Al2 O3 interface up layer-by-layer and using uniform Pd nanocrystals, this work demonstrates the importance of controlled and tunable materials in determining metal-support interactions and catalyst activity.


The use of banquettes of Posidonia oceanica as a source of fiber and minerals in ruminant nutrition. An observational study.
In the search for new food sources that contribute to the optimization of livestock production this paper discusses the possibility of using waste (called banquettes) of a marine plant commonly found on the Mediterranean coasts: Posidonia oceanica. The idea stems from the use of a waste that in summertime generates large costs because it is considered bothersome on the beaches. Thus, tons and tons of residues are collected each year from the beach, being destined for incineration. However, alternative uses for these residues are suggested, such as forage that is particularly relevant for the Mediterranean coast, where the weather does not support abundant grass growth. With this purpose, samples of banquettes of P. oceanica from six different points of a touristic place located in the Murcia Region (S.E. of Spain) were collected in April 2012 on the same day directly from the beach above the water line, washed with distilled water and sun-dried for 48 h. Approximately 500 g of each sample of plant material was chopped and two subsamples of 200 g each were placed in airtight plastic containers and sent to the laboratory for mineral and chemical analysis. This report provides data on the nutritional composition of P. oceanica such as mineral contents (As, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn) and chemical composition (ash, CP, ether extract (EE), NDF and ADF, respectively) and ADL. Finally, the in sacco rumen disappearance of dry matter (DM), organic matter (OM), CP and NDF were studied. Our results showed that minerals, except for Fe, where within the range of values reported for P. oceanica in other parts of the Mediterranean region. Given the high Fe content further studies assessing the antagonic Fe-Cu interaction and its effect on animal health should be addressed. In relation to chemical composition, it is clear that this seagrass is a poor protein source and has levels of DM degradability at 24 h, similar to those obtained for cereal straw. The information summarized here shows some potential for the use of banquettes of P. oceanica as forage source in ruminant nutrition.


What Cross-morphemic Letter Transposition in Derived Nonwords Tells us about Lexical Processing.
According to an obligatory decomposition account of polymorphemic word recognition, a nonword that is composed of a real word plus derivational affix (e.g., teachen) should prime its stem (TEACH) to the same extent that a truly suffixed word does (e.g., teacher). The stem will be activated in both cases after the suffix is removed prior to the lexical status of the letter-string being of relevance. Importantly, disruption to the stem and suffix through letter transposition should have the same impact on the nonwords and words, with teacehn and teacehr equally priming TEACH. However, an experiment by Diependaele, Morris, Serota, Bertrand, and Grainger (2013) found that the equivalent priming for nonwords and words only occurred when they were intact. When letters were transposed, only the truly derived words showed priming. Since such a result cannot be handled by an obligatory decomposition account, it is important to replicate it. Therefore, the present study repeated the conditions of Diependaele et al. (2013), along with a nonword condition where the stem was followed by a non-suffix (e.g., teachin or teacihn). It was found that priming was maintained across all conditions regardless of letter transposition, hence maintaining obligatory decomposition as a viable account. However, the findings with the non-suffixed nonwords led to the conclusion that morphological structure does not control decomposition, but rather, has its impact after form-based components of the letter-string have been activated.


Developmental changes in effective connectivity associated with relational reasoning.
Rostrolateral prefrontal cortex (RLPFC) is part of a frontoparietal network of regions involved in relational reasoning, the mental process of working with relationships between multiple mental representations. RLPFC has shown functional and structural changes with age, with increasing specificity of left RLPFC activation for relational integration during development. Here, we used dynamic causal modeling (DCM) to investigate changes in effective connectivity during a relational reasoning task through the transition from adolescence into adulthood. We examined fMRI data of 37 healthy female participants (11–30 years old) performing a relational reasoning paradigm. Comparing relational integration to the manipulation of single relations revealed activation in five regions: the RLPFC, anterior insula, dorsolateral PFC, inferior parietal lobe, and medial superior frontal gyrus. We used a new exhaustive search approach and identified a full DCM model, which included all reciprocal connections between the five clusters in the left hemisphere, as the optimal model. In line with previous resting state fMRI results, we showed distinct developmental effects on the strength of long-range frontoparietal versus frontoinsular short-range fixed connections. The modulatory connections associated with relational integration increased with age. Gray matter volume in left RLPFC, which decreased with age, partly accounted for changes in fixed PFC connectivity. Finally, improvements in relational integration performance were associated with greater modulatory and weaker fixed PFC connectivity. This pattern provides further evidence of increasing specificity of left PFC function for relational integration compared to the manipulation of single relations, and demonstrates an association between effective connectivity and performance during development.


The association of chronic atrial fibrillation with right atrial dilatation and left ventricular dysfunction in the elderly.
Left atrial dilatation is often considered as one of the important causes for atrial fibrillation. In this study we sought to examine the relationship between right atrial dilatation and left ventricular function in patients with atrial fibrillation, and the association with the documented duration of the dysrrhythmia. 56 consecutive patients with atrial fibrillation were investigated by means of clinical history, electrocardiography and echocardiography. Right atrial dilatation was found in 34, left atrial dilatation in 36 and bi-atrial dilatation in 31 patients. Patients with a dilated right atrium had a larger left atrium, lower left ventricular shortening fraction, and higher transmitral flow velocity than those with a normal right atrium. A history of atrial fibrillation of over 6 months was associated with enlarged atria, reduced left ventricular shortening fraction and increased transmitral flow compared to that of 3 months or less. Functional mitral and tricuspid regurgitation were only found in patients whose atrial fibrillation was over 6 months in duration. Dilatation of both right and left atria is common in chronic atrial fibrillation, and is associated with impaired left ventricular function. A longer duration of atrial fibrillation predisposes to atrial dilatation, left ventricular dysfunction, and functional atrio-ventricular regurgitation. These findings suggest that atrial fibrillation may have a significant contribution to morphological and functional cardiac changes, and raise the possibilities that early cardioversion or adequate rate control might prevent these changes and may improve prognosis in the elderly.


Clinician factors related to outcome differences between black and white patients at CMHCs.
Data are limited on how clinicians contribute to outcome differences between black patients and white patients. Because the clinician-patient relationship is the foundation of mental health services, understanding clinicians' role in outcome differences may help identify evidence-based interventions that decrease disparities and capitalize on positive differences. Symptoms and functioning in a sample of black and white adults receiving outpatient services were examined to determine the effects of their primary clinician on those patterns. The study included 551 patients (25% black) with serious mental illness and 62 mental health professionals (21% black) identified as the patients' primary clinician. Treatment outcomes were measured at baseline and two follow-ups (two and four months) with the Behavior and Symptom Identification Scale, a measure of symptoms and functioning. Data were analyzed with hierarchical linear modeling. Clinicians' levels of multicultural competence, burnout, and education were analyzed. Clinicians moderated the relationship between patient race and outcome differences. There was significant variability among clinicians: approximately 20% had black patients whose outcomes were worse than those of their white patients, and 40% had black patients with better outcomes than their white patients. The only clinician factor predicting these differences was clinician's general experiences and relationships with people from racial-ethnic and cultural groups other than their own. The occurrence of outcome differences varied across clinicians, with some clinicians magnifying outcome differences between black and white patients and others minimizing them. Factors other than clinicians' race, multicultural competence, education, and burnout may contribute to outcome differences between black and white patients.


A randomized control trial of a multiplex gastrointestinal PCR panel versus usual testing to assess antibiotics use for patients with infectious diarrhea in the emergency department.
This study analyzed physician treating behavior through the use of a multiplex gastrointestinal polymerase chain reaction (GI PCR) test compared with usual testing in emergency department (ED) patients with suspected acute infectious diarrhea to assess differences in antibiotic management. A prospective, single-center, randomized control trial was designed to investigate antibiotic use in ED patients with moderate to severe suspected infectious diarrhea, comparing those who received GI PCR to those who received usual testing. ED patients with signs of dehydration, inflammation, or persistent symptoms were randomized to either the experimental arm (GI PCR) or the control arm (usual testing or no testing). A total of 74 patients met study criteria and were randomized to either the experimental GI PCR arm (n = 38) or to the control arm (n = 36). Participants in the GI PCR arm received antibiotics in 87% of bacterial or protozoal diarrheal infections (13/15) whereas those in the control arm received antibiotics in 46% of bacterial or protozoal infections (6/13) (P value 0.042) with 2-proportion difference 0.41 (95% confidence interval 0.07 and 0.68). ED use of multiplex GI PCR led to an increase in antibiotic use for bacterial and protozoal causes of infectious diarrhea compared to usual testing. This increase in antibiotics appears to be appropriate given patients' moderate to severe symptoms and a definitive identification of a likely bacterial or protozoal cause of symptoms. Results should be interpreted with caution because of the small sample size.


Considering long-term sustainability in the development of world class success.
The developmental practice patterns leading to the highest levels of success remain a subject of debate. The present study purposes to extend the body of empirical research by analysing athletic biographies from a large sample of German national squad athletes across all Olympic sports (n=1558; 57% male, 43% female). In a combined retrospective and longitudinal study utilising postal questionnaires, we evaluated the age at onset, volume, domain-specificity, variability in training and competition and success attained at different ages. Developmental practice patterns leading to rapid adolescent success and long-term senior success were inconsistent, and in some aspects contradictory. An early start-age for training and competition, early specialisation, high-intensity specific practice in the respective domain sport and little or no involvement in other sports (OS) favoured early adolescent success. Juvenile success, however, did not contribute to individual differences in success achieved at a senior age (-0.09<r s <0.03). Senior world class performers differed from national class athletes in a later age for onset of training and competition in their domain sport, later specialisation (14.4 vs. 12.1 years), more involvement in OS (training 66% vs. 51%; competitions 53% vs. 39%), but not in practice volume in their domain sport at any age. Findings were confirmed with longitudinal testing and were widely consistent across types of sports. These findings are interpreted relative to correspondence to deliberate practice and DMSP frameworks while drawing on the concept of long-term sustainability.


Screening for ketosis using multiple logistic regression based on milk yield and composition.
Multiple logistic regression was applied to milk yield and composition data for 632 records of healthy cows and 61 records of ketotic cows in Hokkaido, Japan. The purpose was to diagnose ketosis based on milk yield and composition, simultaneously. The cows were divided into two groups: (1) multiparous, including 314 healthy cows and 45 ketotic cows and (2) primiparous, including 318 healthy cows and 16 ketotic cows, since nutritional status, milk yield and composition are affected by parity. Multiple logistic regression was applied to these groups separately. For multiparous cows, milk yield (kg/day/cow) and protein-to-fat (P/F) ratio in milk were significant factors (P<0.05) for the diagnosis of ketosis. For primiparous cows, lactose content (%), solid not fat (SNF) content (%) and milk urea nitrogen (MUN) content (mg/dl) were significantly associated with ketosis (P<0.01). A diagnostic rule was constructed for each group of cows: (1) 9.978 × P/F ratio + 0.085 × milk yield <10 and (2) 2.327 × SNF - 2.703 × lactose + 0.225 × MUN <10. The sensitivity, specificity and the area under the curve (AUC) of the diagnostic rules were (1) 0.800, 0.729 and 0.811; (2) 0.813, 0.730 and 0.787, respectively. The P/F ratio, which is a widely used measure of ketosis, provided the sensitivity, specificity and AUC values of (1) 0.711, 0.726 and 0.781; and (2) 0.678, 0.767 and 0.738, respectively.


Does the Severity of Foraminal Stenosis Impact Outcomes of Lumbar Decompression Surgery?
To establish the relationship between the magnitude of foraminal stenosis and 1) improvement in patient-reported outcomes, 2) improvement in motor function after lumbar decompression surgery, and 3) difference in surgical outcomes. Patients who underwent one-level posterior lumbar decompression for radiculopathy were retrospectively identified. Patient demographics and surgical characteristics were collected through a query search and manual chart review of the electronic medical records. Foraminal stenosis was determined on magnetic resonance imaging and graded using Lee et al.'s validated methodology as none, mild, moderate, or severe. Surgical outcomes, motor function, and patient-reported outcome measures (PROMs) were compared based on the amount of stenosis (mild vs. moderate vs. severe). Bivariant and multivariant analyses were performed. Severe stenosis demonstrated more 90-day readmissions (0.00% vs. 0.00% vs. 8.57%, respectively, P = 0.019), though this effect did not remain significant on multivariate analysis (P = 0.068). There was no association between stenosis severity and the degree of functional impairment or PROMs preoperatively. Patients with moderate or severe preoperative foraminal stenosis showed improvement in all PROMs after surgery (P < 0.05) except the mental component of the Short Form 12 survey. Notably, central stenosis grade was insignificantly different between groups (P = 0.358). Multivariable logistic regression analysis did not identify any significant independent predictors of surgical outcomes or changes in PROMs. We demonstrated that regardless of foraminal stenosis severity preoperatively, patients have a similar improvement in PROMs, surgical outcomes, and restoration of motor function after lumbar decompression surgery for radiculopathy.


The osmolality of caecal contents in piglets following weaning.
Pigs weaned for five days had caecal contents with significantly lower osmolalities than those of unweaned animals of the same age. Supplementation of a standard weaning diet with therapeutic levels of oxytetracycline to suppress the normal large intestinal microbial flora did not significantly further reduce osmolarity of caecal contents after weaning. This observation suggests that microbial activity in the large intestine may not be sufficiently developed by five days after weaning to have a major influence on absorptive processes at that site. Incomplete development of the large intestinal microflora may be one more factor which acts to predispose the young pig to diarrhoea after weaning.


Suppression and restoration of male fertility using a transcription factor.
The Arabidopsis AtMYB103 gene codes for an R2R3 MYB domain protein whose expression is restricted to the tapetum of developing anthers and to trichomes. Down-regulation of expression using anti-sense leads to abnormal tapetum and pollen development, although seed setting still occurs (Higginson, T., Li, S.F. and Parish, R.W. (2003) AtMYB103 regulates tapetum and trichome development in Arabidopsis thaliana. Plant J. 35, 177-192). In this study, we show that blocking the function of the AtMYB103 gene, employing either an insertion mutant or an AtMYB103EAR chimeric repressor construct under the control of the AtMYB103 promoter, results in complete male sterility and failure to set seed. These plants exhibit similar abnormalities in tapetum and pollen development, with the tapetum becoming highly vacuolated at early stages and degenerating prematurely. No exine is deposited on to the pollen wall. The degeneration of pollen grains commences prior to pollen mitosis, the pollen collapsing and largely lacking cytoplasmic content. A restorer containing the AtMYB103 gene under the control of a stronger anther-specific promoter was introduced into pollen donor plants and crossed into the male sterile plants transgenic for the repressor. The male fertility of F1 plants was restored. The chimeric repressor and the restorer constitute a reversible male sterility system which could be adapted for hybrid seed production. This is the first reversible male sterility system targeting a transcription factor essential for pollen development. Strategies for generating inducible male sterility and maintainable male sterility for the production of hybrid crops are discussed.


The enhancing effect of bone marrow cells on the primary immune response of the isolated perfused spleen.
The addition of bone marrow cells or peripheral lymphocytes to the isolated pig spleen markedly enhanced the primary antibody response after 3-day perfusion and antigenic challenge in vitro. The splenic preparation without added cells or with the addition of marrow cells to an irradiated spleen gave a limited response. Contributory evidence is provided that at least two distinct cell types are needed for antibody production. For optimal antibody response by an isolated perfused spleen, marrow cells or peripheral lymphocytes should be added to the system.


Membrane potential changes during the first steps of coliphage infection.
Immediately after adsorption, phages T4 and T5 induce a partial depolarization of the host cytoplasmic membrane. Infected bacteria respond to this phage-induced effect by a repolarization that leads to a new steady state of reduced membrane potential. The rate and extent of repolarization are adjusted to the intensity of depolarization, which depends on the number of adsorbed phages. Consequently, the new steady state membrane potential is attained in the same interval of time regardless of the maximum depolarization. These membrane potential changes appear to be independent of phage-specific properties (type of phage, presence of DNA and internal proteins, injection process) and of several membrane-related parameters (temperature, external pH, preinfectious level of membrane potential). We propose that phage adsorption to the outer membrane triggers the emission of a signal that is transmitted to the cytoplasmic membrane. Additivity of independent signals is possible when stimuli (phages) are added at the same time. Additional adsorption of phages has no further depolarizing effect as soon as the repolarization begins. We propose that this refractoriness to secondary depolarization nd the shut-off of the first depolarization are induced by the same chemical modification also initiated by adsorption of the first phage.


A classifier for the SNP-based inference of ancestry.
Ancestral inference from DNA could serve as an important adjunct for both standard and future human identity testing procedures. However, current STR methods for the inference of ancestral affiliation have inherent statistical and technical limitations. In an effort to identify bi-allelic markers that can be used to infer ancestral affiliation from DNA, we screened 211 SNPs in the human pigmentation and xenobiotic metabolism genes. Allele frequencies of 56 SNPs (most from pigmentation genes) were dramatically different between groups of unrelated individuals of Asian, African, and European descent, and both observed and simulated log likelihood ratios revealed that the markers were of exceptional value for ancestral inference. Log likelihood ratios of the multilocus estimates of biological ancestry (EAE/EBA) ranged from 7 to 10, which are on par with the best of the STR batteries yet described. A linear classification method was developed for incorporating these SNPs into a classifier model that was 99, 98, and 100% accurate for identifying individuals of European, African, and Asian descent, respectively. The methods and markers we describe are therefore an important first step for the development of a practical multiplex test for the inference of ancestry in a forensics setting.


A Systematic Review Protocol Investigating Community Gardening Impact Measures.
Existing community gardening research has tended to be exploratory and descriptive, utilising qualitative or mixed methodologies to explore and understand community garden participation. While research on community gardening attracts growing interest, the empirical rigour of measurement scales and embedded indicators has received comparatively less attention. Despite the extensive body of community gardening literature, a coherent narrative on valid, high quality approaches to the measurement of outcomes and impact across different cultural contexts is lacking and yet to be comprehensively examined. This is essential as cities are becoming hubs for cultural diversity. Systematic literature reviews that explore the multiple benefits of community gardening and other urban agriculture activities have been undertaken, however, a systematic review of the impact measures of community gardening is yet to be completed. This search protocol aims to address the following questions: (1) How are the health, wellbeing, social and environmental outcomes and impacts of community gardening measured? (2) What cultural diversity considerations have existing community garden measures taken into account? Demographic data will be collected along with clear domains/constructs of experiences, impacts and outcomes captured from previous literature to explore if evidence considers culturally heterogeneous and diverse populations. This will offer an understanding as to whether community gardening research is appropriately measuring this cross-cultural activity.


Histological background of dedifferentiated solitary fibrous tumour.
Dedifferentiation is a histological phenomenon characterised by abrupt transition of histology to a sarcomatous component with high-grade malignant potential in solitary fibrous tumour (SFT). The authors histologically reviewed SFT cases to reveal the histological background of dedifferentiated SFTs. Clinicopathological and histopathological findings of 145 SFT cases were reviewed. Immunohistochemical staining and genetic analysis were also performed. The non-dedifferentiated components showed a cellular component in 45 of 145 (31%), high mitotic rate (≥4/10 high-powered field) in 12 of 145 (8.2%) tumours, necrosis in 7 of 145 (4.8%) tumours, multinodular growth pattern in 39 of 132 (29.5%) available tumours and intratumoural fibrous septa in 37 of 131 (28.2%). Immunohistochemically, the non-dedifferentiated components were positive for CD34 in 128 of 141 (90.7%), bcl-2 in 101 of 133 (75.9%), nuclear pattern of β-catenin in 64 of 127 (50.3%) and p16 in 22 of 140 (15.7%). Loss of Rb protein expression was detected in 17 of 110 (15.4%) cases. Statistically, cellular component, multinodular structure, p16 overexpression and Rb protein loss were significantly associated with dedifferentiation. Moreover, cellular component and multinodular structure were significantly associated with p16 overexpression and Rb protein loss. All the non-deddifferentiated components showed wild type of p53 expression. The dedifferentiated components of all 10 dedifferentiated tumours presented positivity for p16 in 9 of 10 (90%) and mutational type of p53 in 5 of 10 (50%). Loss of Rb protein expression was detected in 6 of 10 (60%). The authors propose that cellular or multinodular transformation may be associated with dedifferentiation. They also suggest that cellular and multinodular transformation may be associated with p16 overexpression and Rb downregulation.


Progress in Characterizing the Human Exposome: a Key Step for Precision Medicine.
Most diseases result from the complex interplay between genetic and environmental factors. The exposome can be defined as a systematic approach to acquire large data sets corresponding to environmental exposures of an individual along her/ his life. The objective of this contribution is to raise awareness within the health informatics community about the importance of dealing with data related to the contribution of environmental factors to individual health, particularly in the context of precision medicine informatics. This article summarizes the main findings after a panel organized by the International Medical Informatics Association - Exposome Informatics Working Group held during the last MEDINFO, in Lyon (France) in August 2019. The members of our community presented four initiatives (PULSE, Digital exposome, Cloudy with a chance of pain, Wearable clinics), providing a detailed view of current challenges and accomplishments in processing environmental and social data from a health research perspective. Projects illustrate a wide range of research methods, digital data collection technologies, and analytics and visualization tools. This reinforces the idea that this area is now ready for health informaticians to step in and contribute their expertise, leading the application of informatics strategies to understand environmental health problems. The featured projects illustrate applications that use exposome data for the investigation of the causes of diseases, health care, patient empowerment, and public health. They offer a rich overview of the expanding range of applications that informatics is finding in the field of environmental health, with potential impact in precision medicine.


Chondrogenic effect of precartilaginous stem cells following NLS-TAT cell penetrating peptide-assisted transfection of eukaryotic hTGFβ3.
Cell penetrating peptides (CPPs) are a series of promising carriers for delivering exogenous DNA to living cells. Among them, the combination of the human immunodeficiency virus TAT protein (TAT) with the SV40 large T protein nuclear localization signal (NLS) to form NLS-TAT performs well. In the present study, we took advantage of this new carrier to deliver transforming growth factor-beta 3 (TGFβ3) genes. TGFβ3 was expressed by the pEGFP-N1 vector following transfection of rat precartilaginous stem cells (PSCs), which promoted hTGFβ3 protein self-expression. At 24, 48, 72, and 120 h after transfection, the expression levels of hTGFβ3 were found to be elevated as compared with the control. The expression of hTGFβ3 was found to mediate the chondrogenic effect of PSCs. Thus, we determined the expression of the chondrogenesis-related genes type II collagen, Sox 9, and aggrecan in PSCs at 24, 48, 72, and 120 h after transfection. We found that their transcription and translation was augmented, which indicated a trend of active chondrogenesis in the PSCs. Our results demonstrated that NLS-TAT had the ability to deliver exogenous DNA into rat PSCs and could be actively expressed. This process successfully promoted PSC chondrogenesis. Additionally, PSC, may represent a new type of stem cells, and thus show great potential in regenerative repair following cartilage injury.


Microleakage of bonded amalgam restorations: effect of thermal cycling.
This study examined the effect of thermal cycling on the microleakage of bonded amalgam restorations. Three dental amalgam alloys and a gallium alloy were tested with two adhesive resin systems and copal varnish as a control. Class V cavity preparations were prepared on 168 freshly extracted premolars or molars. The preparations were placed parallel to and 1.0 mm occlusal to the cementoenamel junction (CEJ). Four groups of 42 teeth each were treated with one of the following adhesive dentin systems: Bond-It, All-Bond 2/Resinomer or a copal varnish (Copalite). The four groups of 42 teeth each were then restored with one of three dental amalgams: Orosphere Plus, Indiloy, Oralloy or a Gallium alloy (Galloy), resulting in 12 test groups of 14 teeth each. The specimens were stored in double distilled water at 37 degrees C for 24 hours. Final contouring and polishing of the restorations were performed under water spray. Half of the restorations in each group were thermocycled for 3000 cycles (5 degrees C-37 degrees C-55 degrees C-37 degrees C) with a dwell time of 15 sec at each temperature. The other half were stored in double distilled water at 37 degrees C for 24 hours. Then all 168 restorations were stained with dye, sectioned and scored for microleakage. Results showed that the adhesive dentin systems reduced microleakage in amalgam restorations compared to copal varnish only in non-thermocycled specimens. Statistical analysis of the results showed that there was an extremely significant difference (p < 0.001) in microleakage between the non-thermocycled and the thermocycled specimens in all test groups, whereas, there was no significant difference (p > 0.05) among thermocycled specimens. The reduction of microleakage was not significantly different between Bond-It and All-Bond 2/Resinomer in non-thermocycled specimens. Oralloy showed the most microleakage in the non-thermocycled groups when compared to the other alloys using the same adhesive liner.


[Psychosocial stress among health care workers].
Health care workers are exposed to high stress levels and psychosocial risks. The imbalance between the invested efforts and received rewards acquires special importance in this setting. To assess the psychosocial risk level and its relationship with depression, distress and psychotropic drug use among health care workers. Seven hundred eighty two workers (602 females; 180 males) answered self-administered questionnaires to measure psychosocial risk and mental health. Twenty five percent of respondents used psychotropic drugs, 34% had a high level of distress and 23% had depressive symptoms. They also reported a low level of decisional latitude (48%), high emotional demands (47%), low social support (41%) and a significant effort-reward imbalance (67%). Those exposed to job strain (high demands and low decisional latitude), iso-strain (job strain plus low social support), and effort-reward imbalance were twice as likely to suffer symptoms of depression and elevated distress compared to non-exposed subjects. There are high levels of psychosocial stress among health care workers.


Enhanced recovery program (ERP) in major laryngeal surgery: building a protocol and testing its feasibility.
Enhanced recovery programs (ERP) represent a multimodal approach to perioperative patient care. The benefits of ERP are well demonstrated in colorectal surgery and Enhanced Recovery After Surgery (ERAS®) programs, that epitomise the ERP concept, have being introduced in different specialties, including vascular, gastric, pancreatic, urogynecologic and orthopaedic surgery. However, no ERP has been proposed for head and neck surgery. We developed an expert-opinion-based ERP for laryngeal surgery based on the key principles of colorectal surgery ERAS®. Twenty-four patients undergoing major laryngeal surgery (total and partial laryngectomies or surgical removal of oropharyngeal tumour with muscle flap reconstruction) were treated according to such an ERP protocol, which differed under several respects from our previous standard practice (described in 70 consecutive patients who underwent major laryngeal surgery before ERP implementation. The adherence rate to the different ERP items is reported. Adherence to ERP items was high. Nutritional assessment, antibiotic prophylaxis, postoperative nausea and vomit (PONV) prophylaxis and postoperative speech therapy targets were applied as required in 100% of cases. Some ERP items (antibiotic prophylaxis, intraoperative infusion rate, and postoperative speech therapy) were already frequently implemented before ERP adoption. Postoperative medical complications occurred in 8.3% of patients. Our expert opinion-based ERP protocol for major laryngeal surgery proved feasible. The degree of benefit deriving from its implementation has yet to be assessed.


Octreotide-Paclitaxel Conjugate Reverses Paclitaxel Resistance by p38 Mitogen-Activated Protein Kinase (MAPK) Signaling Pathway in A2780/Taxol Human Ovarian Cancer Cells.
BACKGROUND Platinum plus paclitaxel is a first-line chemotherapy for ovarian cancer. Platinum resistance is a hot topic for many scholars, but drug resistance caused by paclitaxel is also a topic of concern. Currently, scholars believe that inhibition of MAPK signaling pathway may be an effective way to reverse the drug resistance of tumor paclitaxel. MATERIAL AND METHODS A2780/Taxol cells or nude mice were divided into 8 groups: control group, OCT (octreotide) group, OC (octreotide+cyclosomatostatin) group, PTX (paclitaxel) group, PO (paclitaxel+octreotide) group, POC (paclitaxel+octreotide+cyclosomatostatin) group, P-O (octreotide-paclitaxel conjugate) group, and P-OC (octreotide-paclitaxel conjugate+cyclosomatostatin) group. The phosphorylation level of p38 MAPK and the expression level of vascular endothelial growth factor (VEGF) were determined by western blot. Flow cytometry was used to discover the apoptosis of A2780/Taxol cells and xenografts. The expression of class III beta-tubulin was detected by immunohistochemistry. RESULTS Octreotide-paclitaxel conjugate inhibited phosphorylation of the p38MAPK signal pathway, decreased the expression of downstream VEGF, and increased the apoptosis of drug-resistant cancer cells. In addition, it reduced the expression of class III beta-tubulin protein and increase the sensitivity of drug-resistant cells to paclitaxel. All these effects of octreotide-paclitaxel conjugate were cancelled by cyclosomatostatin. CONCLUSIONS Octreotide-paclitaxel conjugate can reverse the paclitaxel resistance of A2780/Taxol human ovarian cancer cells by inhibiting the activity of p38 MAPK signaling pathway.


Studies of intestinal morphology and cathepsin B expression in a transgenic mouse aiming at intestine-specific expression of Cath B-EGFP.
Cathepsin B has been shown to not only reside within endo-lysosomes of intestinal epithelial cells, but it was also secreted into the extracellular space of intestinal mucosa in physiological and pathological conditions. In an effort to further investigate the function of this protease in the intestine, we generated a transgenic mouse model that would enable us to visualize the localization of cathepsin B in vivo. Previously we showed that the A33-antigen promoter could be successfully used in vitro in order to express cathepsin B-green fluorescent protein chimeras in cells that co-expressed the intestine-specific transcription factor Cdx1. In this study an analog approach was used to express chimeric cathepsin B specifically in the intestine of transgenic animals. No overt phenotype was observed for the transgenic mice that reproduced normally. Biochemical and morphological studies confirmed that the overall intestinal phenotype including the structure and polarity of this tissue as well as cell numbers and differentiation states were not altered in the A33-CathB-EGFP mice when compared to wild type animals. However, transgenic expression of chimeric cathepsin B could not be visualized because it was not translated in situ although the transgene was maintained over several generations.


The prevalence and frequency of drug use among Western Australian metropolitan high school students.
Data pertaining to prevalence and frequency of drug use were obtained from 1,394 Western Australian metropolitan high school students using a self-report questionnaire. Alcohol, marijuana, tobacco, hallucinogens, and amphetamines were reported as the most prevalent substances, with over 50% of "current drug users" using alcohol and marijuana on a frequent basis (i.e., weekly to more than once per day). Significant interactions existed between Gender and prevalence of tobacco and hallucinogens; and School Year Level and prevalence of tobacco, alcohol, hallucinogens and amphetamines. In terms of the frequency of use, significant interactions were found between Gender and marijuana; and between School Year Level and tobacco. Approximately 40% of substance-using participants used one single substance, 40% used two or three substances, and 20% used four or more substances. The results suggest there is a need for educators to have a greater understanding of the patterns of substance use in order for them to more aptly shape drug education programs.


Evaluation of dabigatran utilization and risk among hospitalized patients.
Few data exist to evaluate the use of dabigatran among hospitalized patients and this population that may be at increased risk for toxicity or reduced efficacy. As such, the objectives of this study were to describe the characteristics of hospitalized patients prescribed dabigatran and to compare select characteristics with patients included in the Randomized Evaluation of Long-Term Anticoagulation (RE-LY) phase III clinical trial. A cross-sectional study was performed at Upstate University Hospital. Select patient characteristics, including age, renal function, and drug interaction exposure, were compared with those for patients in the RE-LY trial. The study included 140 patients. Mean age was 69.7 years (SD = 13.8) and 72 (51.4%) were male. The mean CHADS2 score was 2.48 as compared with 2.2 in RE-LY. Significantly more patients in our study had moderate to severe renal dysfunction (creatinine clearance [CrCl] < 50 mL/min) as compared with the RE-LY trial (27.5% vs 19.4%, P = .0207) when analyzing our study patients' worst recorded CrCl. Among the 29 patients prescribed dose-adjusted dabigatran, 3 were correctly dose adjusted, 2 were overdosed, and 26 were underdosed. At least 1 pharmacokinetic drug interaction with dabigatran occurred in 110 study patients (78.6%). Use of proton pump inhibitors was more predominant in our study population when compared with RE-LY (64.3% vs 13.9%, P < .0001). Hospitalized patients exhibit multiple characteristics that place them at heightened risk for altered dabigatran drug concentrations and may have a heightened risk for clinical sequelae related to dabigatran use.


Hardware-Efficient 1D CNN for Patient-Specific Early Seizure Detection.
Closed-loop brain-implantable neuromodulation devices are a new treatment option for patients with refractory epilepsy. Seizure detection algorithms implemented on such devices are subject to strict power and area constraints. Deep learning methods, though very powerful, tend to have high computational complexity and thus are typically impractical for resource-constrained neuromodulation devices. In this paper, we propose a compact and hardware-efficient one-dimensional convolutional neural network (1D CNN) structure for patient-specific early seizure detection. Feature extraction techniques and a novel initialization method based on the forward-chaining training and testing scheme are used to improve model performance. Our compact model achieves similar accuracy to that of support vector machines, the state-of-the-art method for seizure detection, while consuming over 20x less power.


Proton pump inhibitor influence on reflux in Barrett's oesophagus.
It is important to identify factors responsible for the development of Barrett's oesophagus (BO). The effect of proton pump inhibitors (PPIs) on oesophageal clearance of both acid and alkaline reflux in these patients is uncertain and studies comparing BO patients and healthy controls (HCs) have not been performed earlier. Two groups of patients were studied: 18 HCs and 12 BO patients. Oesophageal motility, acid reflux and duodenogastro-oesophageal reflux (DGOR) were measured using a three-pressure transducer catheter with an antimony pH tip, connected to a sodium ion selective electrode. All patients were studied both on and off PPIs. Without PPI therapy, BO patients had significantly more upright and supine acid reflux and upright DGOR compared with HCs. During acid reflux, HC demonstrated more peristalsis than BO [HC, % peristalsis=64 (9), BO=53 (8), P<0.01], but this was not seen during DGOR. [HC, % peristalsis=68 (14), BO=56 (11)]. In Barrett's patients, DGOR was significantly reduced with PPIs [off PPI, % upright DGOR=61 (17), on PPIs=19 (15), P<0.01], and no oesophageal motility differences were seen compared with results without PPIs. HCs demonstrate better oesophageal motility compared with BO patients to prevent acid and alkaline reflux. When acid reflux occurred, HCs had better coordinated motility to remove it. This increased coordination did not occur during DGOR, suggesting different stimulation mechanisms. PPI reduced DGOR in BO patients, without any change in oesophageal motility.


[Prevention of complications of ovarian stimulation in in-vitro fertilization].
The ovarian hyperstimulation syndrome which is the most severe iatrogenic complication of ovulation induction, may range from mere abdominal discomfort to lethal thromboembolism. It is therefore, mandatory to be able to evaluate the risk of such complications. In in vitro fertilization treatment, maximal level and growth rate of oestradiol and also the number of oocytes retrieved, are significantly higher among patients who will develop an hyperstimulation syndrome than in a group of healthy controls. However, due to the large distribution of individual values, none of these variables can be predictive by itself. Only multiple discriminant analysis effected within the frame of a multicentric study might be able to define a specific and sensitive predictive function.


MELANOMA IN GOLDEN AND KING WILDEBEESTS ( CONNOCHAETES TAURINUS).
Pigmentation of the skin is a crucial component in the pathogenesis of melanocytic neoplasms and other skin-related tumors, as melanin is known to function in both the absorbance of ultraviolet radiation and as an antioxidant. Very limited information exists regarding the incidence and metastatic potential of neoplastic conditions of the skin in game animals, especially wildebeests, relative to domestic animals. Four cases of cutaneous melanoma in color-variant golden and king wildebeests ( Connochaetes taurinus) (from 2014 to 2015) in South Africa were investigated. Melanoma in these captive animals was characterized using histopathology, transmission electron microscopy, and an immunohistochemistry panel, which consisted of monoclonal antibodies against three melanocytic markers: Melan A, PNL2, and S100. Overall, 2/4 cases (50%) of the melanocytic neoplasms stained strongly positive for all the melanocytic markers, while 4/4 cases (100%) stained positively for at least one of the markers. Cutaneous melanocytic neoplasia has not been reported in wildebeests; the current study suggests that selection of wildebeests for coat color potentially predisposes to this condition.


A dimer of BPV-1 E2 containing a protease resistant core interacts with its DNA target.
The E2 proteins encoded by papillomaviruses interact with the viral DNA to regulate its transcription. In the present study, we have constructed bacterial vectors expressing the full-length or N-terminal truncated BPV-1 E2 proteins under the control of an inducible promoter. By UV cross-linking experiments we show that a dimer of the intact or truncated E2 protein interacts with a single palindromic site ACCGNNNNCGGT. The DNA-binding domain of E2 can be reduced to a small protease resistant core. Methylation interference studies show that this C-terminal domain interacts with the major groove of the DNA by contacting two consecutive guanine residues in both halves of the palindrome. Although one binding site is sufficient for high affinity binding in vitro or in vivo, two E2 binding sites are required for transcriptional activation in eukaryotic cells.


Reactive oxygen species and nitric oxide imbalances lead to in vivo and in vitro arrhythmogenic phenotype in acute phase of experimental Chagas disease.
Chagas Disease (CD) is one of the leading causes of heart failure and sudden death in Latin America. Treatments with antioxidants have provided promising alternatives to ameliorate CD. However, the specific roles of major reactive oxygen species (ROS) sources, including NADPH-oxidase 2 (NOX2), mitochondrial-derived ROS and nitric oxide (NO) in the progression or resolution of CD are yet to be elucidated. We used C57BL/6 (WT) and a gp91PHOX knockout mice (PHOX-/-), lacking functional NOX2, to investigate the effects of ablation of NOX2-derived ROS production on the outcome of acute chagasic cardiomyopathy. Infected PHOX-/- cardiomyocytes displayed an overall pro-arrhythmic phenotype, notably with higher arrhythmia incidence on ECG that was followed by higher number of early afterdepolarizations (EAD) and 2.5-fold increase in action potential (AP) duration alternans, compared to AP from infected WT mice. Furthermore, infected PHOX-/- cardiomyocytes display increased diastolic [Ca2+], aberrant Ca2+ transient and reduced Ca2+ transient amplitude. Cardiomyocyte contraction is reduced in infected WT and PHOX-/- mice, to a similar extent. Nevertheless, only infected PHOX-/- isolated cardiomyocytes displayed significant increase in non-triggered extra contractions (appearing in ~75% of cells). Electro-mechanical remodeling of infected PHOX-/-cardiomyocytes is associated with increase in NO and mitochondria-derived ROS production. Notably, EADs, AP duration alternans and in vivo arrhythmias were reverted by pre-incubation with nitric oxide synthase inhibitor L-NAME. Overall our data show for the first time that lack of NOX2-derived ROS promoted a pro-arrhythmic phenotype in the heart, in which the crosstalk between ROS and NO could play an important role in regulating cardiomyocyte electro-mechanical function during acute CD. Future studies designed to evaluate the potential role of NOX2-derived ROS in the chronic phase of CD could open new and more specific therapeutic strategies to treat CD and prevent deaths due to heart complications.


FOXO1 deficiency impairs proteostasis in aged T cells.
T cell differentiation involves the dynamic regulation of FOXO1 expression, which rapidly declines after activation and is subsequently restored. Reexpression is impaired in naïve CD4+ T cell responses from older individuals. Here, we show that FOXO1 promotes lysosome function through the induction of the key transcription factor for lysosomal proteins, TFEB. Subdued FOXO1 reexpression in activated CD4+ T cells impairs lysosomal activity, causing an expansion of multivesicular bodies (MVBs). Expansion of the MVB compartment induces the sequestration of glycogen synthase kinase 3β (GSK3β), thereby suppressing protein turnover and enhancing glycolytic activity. As a consequence, older activated CD4+ T cells develop features reminiscent of senescent cells. They acquire an increased cell mass, preferentially differentiate into short-lived effector T cells, and secrete exosomes that harm cells in the local environment through the release of granzyme B.


Evaluation of pain relief.
Obtaining objective data on pain-relieving procedures is a simple matter. Measures of pain, activity levels and analgesic intake can be collected not only in a hospital but also in an outpatient setting. The data obtained make possible assessment of the effectiveness of a technique in any particular patient, and when combined for groups of patients, such data facilitate comparison of the effectiveness of different techniques in different pain states.


[Genetic foundations of vulnerability to depression].
Depression is a complex phenotype probably heterogeneous in its biology and in its aetiology. Genetic studies have contributed to clarifying some aspects of that heterogeneity, but we are still far from being able to establish its molecular foundations, although a certain degree of genetic variability in genes of the serotoninergic system appear to contribute to the risk of the disease and to certain of its clinical aspects. The risk of a major depressive episode increases in relation to the number of genes shared with an affected person but, in genetic terms and given the differential risks found, there is also a certain overlap with bipolar disorder and schizophrenia. These data lead to reflection on the meaning of genetic risk detected in families and its specificity. Perhaps genetic studies will have to leave the categorical definition of phenotypes and begin to use the dimensions of psychopathology for delimiting the biological and genetic subgroups present in functional psychoses.


Freeze-dried demineralized bone in dental implant reconstruction.
Rebuilding deficient alveolar bone to assist in reconstruction for dental implants is a frequent task for the oral and maxillofacial surgeon. Freeze-dried demineralized bone is being used with increasing frequency. It is a reliable, safe and inexpensive bone source and has very few limitations. Freeze-dried demineralized bone processing is reviewed and its safety is emphasized to aid the general practitioner in discussing the concepts with prospective patients. Cases are presented showing the most frequent uses of the material.


Ischemia-induced phosphorylation of phospholemman directly activates rat cardiac Na/K-ATPase.
Regulation of the Na/K ATPase by protein kinases is model-specific. We have observed a profound activation of the sarcolemmal Na/K ATPase during cardiac ischemia, which is masked by an inhibitor of the enzyme in the cytosol. The aim of these studies was to characterize the pathways involved in this activation in the Langendorff-perfused rat heart. Na/K ATPase activity was determined by measuring ouabain-sensitive phosphate generation by cardiac homogenates at 37 degrees C. In isolated sarcolemma, ischemia (30 min) caused a substantial activation of the Na/K ATPase compared with aerobic controls, which was abolished by perfusing the heart with staurosporine or H89. However, the alpha1 subunit of the Na/K ATPase was not phosphorylated during ischemia. The sarcolemmal protein phospholemman (PLM) was found associated with the Na/K ATPase alpha1 and beta1 but not alpha2 subunits, and PLM increased its association with the catalytic subunit of PKA following ischemia. In vitro 14-3-3 binding assays indicated that PLM was phosphorylated following ischemia. These results indicate that the ischemia-induced activation of the Na/K ATPase is indirect, through phosphorylation of PLM, which is an integral part of the Na/K ATPase enzyme complex in the heart. The role of PLM is analogous to phospholamban in regulating the sarcoplasmic reticulum calcium ATPase.


Absolute quantification reveals the stable transmission of a high copy number variant linked to autoinflammatory disease.
Dissecting the role copy number variants (CNVs) play in disease pathogenesis is directly reliant on accurate methods for quantification. The Shar-Pei dog breed is predisposed to a complex autoinflammatory disease with numerous clinical manifestations. One such sign, recurrent fever, was previously shown to be significantly associated with a novel, but unstable CNV (CNV_16.1). Droplet digital PCR (ddPCR) offers a new mechanism for CNV detection via absolute quantification with the promise of added precision and reliability. The aim of this study was to evaluate ddPCR in relation to quantitative PCR (qPCR) and to assess the suitability of the favoured method as a genetic test for Shar-Pei Autoinflammatory Disease (SPAID). One hundred and ninety-six individuals were assayed using both PCR methods at two CNV positions (CNV_14.3 and CNV_16.1). The digital method revealed a striking result. The CNVs did not follow a continuum of alleles as previously reported, rather the alleles were stable and pedigree analysis showed they adhered to Mendelian segregation. Subsequent analysis of ddPCR case/control data confirmed that both CNVs remained significantly associated with the subphenotype of fever, but also to the encompassing SPAID complex (p < 0.001). In addition, harbouring CNV_16.1 allele five (CNV_16.1|5) resulted in a four-fold increase in the odds for SPAID (p < 0.001). The inclusion of a genetic marker for CNV_16.1 in a genome-wide association test revealed that this variant explained 9.7 % of genetic variance and 25.8 % of the additive genetic heritability of this autoinflammatory disease. This data shows the utility of the ddPCR method to resolve cryptic copy number inheritance patterns and so open avenues of genetic testing. In its current form, the ddPCR test presented here could be used in canine breeding to reduce the number of homozygote CNV_16.1|5 individuals and thereby to reduce the prevalence of disease in this breed.


Prevalence of positive substance abuse screen results among adolescent primary care patients.
To measure the prevalence of positive substance use screen results among adolescent primary care patients and to estimate the prevalence of substance-related problems and disorders. Cross-sectional survey. A network of primary care practices in New England. A consecutive sample of 12- to 18-year-old patients (N = 2133), with a study participation rate of 92.7%. The CRAFFT substance abuse screening test (a full description of this screen is given in the "Introduction"). Frequencies of positive screen results were computed for the entire sample, each practice, visit type (well-child care, sick visit, follow-up, or other), and patient status (new vs established). Generalized estimating equation modeling was used to test for difference in proportions. CRAFFT scores, demographic data, and Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) diagnostic data from a previous study were used to estimate the prevalence of problematic substance use, abuse, and dependence. Overall, 14.8% of adolescents had positive results on the CRAFFT screen. Prevalence rates differed significantly across practices (P < .001) after adjusting for demographic factors. The highest positive rates on the CRAFFT screen were at school-based health centers (29.5%) and the rural family practice (24.2%), the middle rate was at the adolescent clinic (16.6%), and the lowest rates were at the health maintenance organization (14.1%) and pediatric clinic (8.0%). Sick visits had the highest rate (23.2%). Well-child care visits had a significantly lower rate (11.4%, P < .001). Statistical modeling estimated that 11.3% of all patients had problematic use, 7.1% had abuse, and 3.2% had dependence. Substance abuse screening should occur whenever the opportunity arises, not at well-child care visits only.


Similar mechanisms of traumatic rectal injuries in patients who had anal sex with animals to those who were butt-fisted by human sexual partner.
Sexual pleasure comes in various forms of physical play, for many it involves stimulation of the vagina, while the anus for others; some enjoy both. A recent report by Cappelletti et al.1 shows a meta-analysis of cases describing anal trauma due to sexual fisting in human partners. This clinical article reports four cases of males diagnosed with zoophilia, and who received anal sex from animals, resulting in injuries. Surgical and psychiatric evaluations are summarized. Unusual etiology of sexual activity with animals caused peri-anal trauma in men who engaged in anal sex with dogs and farm animals. Injuries to patients who receive anal sex from animals are mechanistically similar to fisting-induced rectal damage. Among zoophiles, the mode of harm occurs through blood-engorged, interlocked penis that causes tissue lacerations upon retraction from an anus. In people experimenting with fisting, repetitive stretching within anal canal and of external sphincter causes the internal injuries. The mode of physical stimulation explains the extent of injuries in fisters vs. zoophiles: in fisting, the pressure applied by hand is controllable proximally around and within anal sphincter, while penetration by the animal penis is unpredictable and occurs within the proximal anal canal. Forensically, the findings presented in this article describe a significant mechanism of injury in fisters versus passive zoophiles. These descriptions may aid in clinically differentiating pleasurable and pathological rectal stimulation.


Chemotaxis by a CNS macrophage, the microglia.
Microglia demonstrate many characteristics similar to those seen in monocytes and tissue-specific macrophages, including phagocytosis, production of oxygen radicals, and growth factors and expression of MHC antigens. We have examined the ability of microglia, cultured from the cerebral cortices of neonatal rats, to demonstrate another important functional characteristic of monocytic-derived cells, that is, chemotaxis. Our results show that cultured rat microglia demonstrate chemotaxis to complement dependent chemoattractants such as recombinant C5a, zymosan activated serum, and to rat serum as well as to transforming growth factor-beta, a chemoattractant produced by platelets. Microglia fail to migrate to bacterial dependent chemoattractants such as the N-formyl peptides. The failure to respond is not dependent on maturational state of the microglia. Treatment with DMSO or casein, agents known to induce morphological and functional changes in cultured microglia reminescent of a "resting" and an "activated" macrophage, respectively, do not alter the response to fMet-Leu-Phe. In addition, the chemotactic response to serum in DMSO or casein-treated cells is the same as the response seen in untreated day 10 cultured microglia or untreated age-matched controls. The ability of microglia to migrate in response to inflammatory mediators suggests that these cells can move to sites of injury, thereby enabling them to participate in an inflammatory response.


Endometriosis and mammographic density measurements in the Nurses' Health Study II.
Endometriosis and mammographic density have been hypothesized to be influenced by sex steroid hormonal exposures in adolescence and early adulthood. We investigated the association between endometriosis and mammographic density, a consistent and independent risk factor for breast cancer. We conducted a cross-sectional analysis among 1,581 pre- and postmenopausal women not previously diagnosed with breast cancer in the Nurses' Health Study II cohort. We measured average percent mammographic density and absolute dense and non-dense breast area using a validated computer-assisted method. Multivariable linear regression was used to estimate the association between endometriosis and mammographic density among pre- and postmenopausal women separately. Among premenopausal women, average percent mammographic density was 43.1 % among women with endometriosis (n = 91) and 40.5 % among women without endometriosis (n = 1,150). Endometriosis was not associated significantly with mammographic density among premenopausal (% difference = 2.00 percentage points 95 % CI -1.33, 5.33) or among postmenopausal women (% difference = -0.89 percentage points 95 % CI -5.10, 3.33). Among premenopausal women, there was heterogeneity by BMI at age 18 (p value = 0.003), with a suggested association among those who were lean at age 18 (BMI < 20.6 kg/m(2)) (% difference = 3.74 percentage points 95 % CI -0.29, 7.78). Endometriosis was not found to be associated with overall measurements of mammographic density.


MicroRNA-200C and -150 play an important role in endothelial cell differentiation and vasculogenesis by targeting transcription repressor ZEB1.
To investigate the role of miRNA in controlling human embryonic stem (hES) cell differentiation toward the endothelial lineage and chick embryonic blood vessel formation, undifferentiated hES cells were first cultured on Matrigel-coated flasks and in endothelial cell growth medium-2 (EGM-2) to initiate endothelial cell (EC) differentiation. CD146(+) cells were isolated from differentiating hES cells and expanded in vitro. The in vitro expanded CD146(+) cells were positive for EC markers, capable of Ac-LDL uptake, lectin binding, and the formation of vascular structures in vitro and in vivo. miRNA gain/loss-of-function analyses revealed that miR-150 and miR-200c were crucial in EC differentiation. Transcriptional repressor zinc finger E-box-binding homeobox 1 (ZEB1) was identified as the communal target gene of miRNA-200C and -150, and inhibition of ZEB1 was required for miRNA-200C or -150 mediated EC gene expressions. Moreover, we demonstrated that ZEB1 could transcriptionally repress EC gene expression through direct binding to promoters of EC genes. Finally, we also demonstrated that miRNA-200c and -150 played an important role in chick embryonic blood vessel formation by in vivo inhibition of miRNA-200C or -150 in developing chick embryos, and blocking ZEB1 signaling in CD146-positive cells could rescue the inhibitory effects of miR-200c inhibiton in in vivo vasculogenesis. Our findings revealed that miR-150 and miR-200c play an important role in human endothelial lineage specification and chick embryonic vasculogenesis by targeting ZEB1.


The potential role of a self-management intervention for ulcerative colitis: a brief report from the ulcerative colitis hypnotherapy trial.
Inflammatory bowel diseases (IBD) are chronic inflammatory illnesses marked by unpredictable disease flares, which occur spontaneously and/or in response to external triggers, especially personal health behaviors. Behavioral triggers of flare may be responsive to disease self-management programs. We report on interim findings of a randomized controlled trial of gut-directed hypnotherapy (HYP, n = 19) versus active attention control (CON, n = 17) for quiescent ulcerative colitis (UC). To date, 43 participants have enrolled; after 5 discontinuations (1 in HYP) and 2 exclusions due to excessive missing data, 36 were included in this preliminary analysis. Aim 1 was to determine the feasibility and acceptability of HYP in UC. This was achieved, demonstrated by a reasonable recruitment rate at our outpatient tertiary care clinic (20%), high retention rate (88% total), and our representative IBD sample, which is reflected by an equal distribution of gender, an age range between 21 and 69, recruitment of ethnic minorities (∽20%), and disease duration ranging from 1.5 to 35 years. Aim 2 was to estimate effect sizes on key clinical outcomes for use in future trials. Effect sizes (group × time at 20 weeks) were small to medium for IBD self-efficacy (.34), Inflammatory Bowel Disease Questionnaire (IBDQ) total score (.41), IBDQ bowel (.50), and systemic health (.48). Between-group effects were observed for the IBDQ bowel health subscale (HYP > CON; p = .05) at 20 weeks and the Short Form 12 Health Survey Version 2 (SF-12v2) physical component (HYP > CON; p < .05) at posttreatment and 20 weeks. This study supports future clinical trials testing gut-directed HYP as a relapse prevention tool for IBD.


E-cadherin as an indicator of mesenchymal to epithelial reverting transitions during the metastatic seeding of disseminated carcinomas.
Cancer metastasis follows a sequential series of events, and many of the critical steps are distinctly similar to EMT-like transformations that occur during normal embryonic development. A current area of focus is the similarities between how cancer cells interact with the ectopic parenchyma after metastatic spread, and secondary developmental MET events that occur in epithelial tissues that have re-assembled within the embryo from mesenchymal cells. Accumulating evidence suggests a critical role for these secondary events, termed mesenchymal-epithelial transitions (MET) in development and mesenchymal-epithelial reverting transitions (MErT) in cancer. In this situation, metastatic seed cancer cells may inertly become part of the ectopic tissue and therefore surmount the metastatic inefficiencies to which most disseminated cancer cells succumb. Just as a critical EMT event is the downregulation or silencing of E-cadherin, we discuss the role of E-cadherin in cancer-associated MErT at distant metastatic sites and speculate on the implications for the fate of micrometastases that undergo a transition to being E-cadherin positive.