Tag Archives: SCR 7

The following are the supplementary data related to

The following are the supplementary data related to this article.

Conflicts of Interest

Author Contributions

Funding
This work was supported by The Hormel Foundation and National Institutes of Health grants CA166011, CA172457 and R37CA081064. These funding sources played no role in the study design, data collection, data analysis or interpretation, or writing of the report.

Acknowledgments
We acknowledge support from Mayo Clinic Center for Cell Signaling in Gastroenterology (NIDDK P30DK084567). We thank Ms. Cindy Nordyke for the technical assistance in clinical tissue processing and Ms. Nicki Brickman at The Hormel Institute, University of Minnesota for assistance in submitting our manuscript.

Introduction
Hyper-IgG4-associated abnormalities are common denominators for several autoimmune fibroinflammatory diseases that can affect any organ from the salivary glands to the SCR 7 or kidneys, collectively referred to as IgG4-related disease [1]. Characteristic pathological features in various affected sites consist of lymphoplasmacytic infiltration with IgG4-positive plasma cells, storiform fibrosis, and variably elevated levels of IgG4 [1]. Kidney lesions are usually accompanied by tubulointerstitial nephritis [2]. Glomerular lesions, including membranous nephropathy (MN), have been reported less frequently [2,3].
We described a 54-year-old male patient with IgG4-related disease manifesting as pancreatitis, Mikulicz disease that later developed nephrotic-range proteinuria, and MN [4] with glomerular lesions partly different from the idiopathic form. Indeed the predominant immunoglobulin deposited in the renal tissue was IgG3, while staining for IgG4 was weak in the absence of circulating anti-phospholipase A2 receptor (PLA2R) antibodies, possibly implying a distinct process. Moreover, no detectable immune complexes were found in the patient\’s serum [4]. On the other hand, we found IgG3 reactivity against superoxide-dismutase-2 (SOD2) [4], a mitochondrial antioxidant previously identified as an autoimmune target in patients with idiopathic MN [5]. The first aim of this study was to clarify the role of IgG4 antibodies in our patient\’s renal disease, and cellular mechanisms underlying SOD2 enrichment on the podocyte plasma membrane. We also wanted to establish whether IgG4-related disease and MN development have a common pathogenic event. Addressing those questions in our patient and in four other IgG4-related disease patients without MN, allowed us to recapitulate the pathogenetic chain of events taking advantage of in vitro disease modeling. Here we propose a two-stage model in which IgG4 anti-carbonic anhydrase II (CAII), an autoantigen candidate in IgG4-related disease patients, is critical for altering pH homeostasis, mitochondrial dynamic, and SOD2 corticalization. At a later stage, mislocated SOD2 serves as a target for the binding of IgG3-subtype autoantibodies capable of fixing complement and amplifying podocyte injury, which contribute to the MN lesion, likely favored by individual genetic predisposition.

Methods

Results

Discussion
The majority of patients with IgG4-related disease, particularly when associated with autoimmune pancreatitis, have circulating IgG4 subclass antibodies that recognize CAII [13,14]. The patient with hyper-IgG4 described here does indeed have both circulating IgG4 reacting with CAII and MN, and we disclose a functional link between the binding of the patient\’s IgG4 to podocytes and the intracellular events underlying the development of the MN lesion. CAII is expressed in most segments of the human kidney, including proximal and distal tubules [10]. In addition to its typical cytosolic distribution, CAII has been found to localize close to the plasma membrane where it interacts with the electroneutral Cl−/HCO3− anion exchanger 1, giving rise to a transport metabolon wherein HCO3− is transferred from CAII to the anion exchanger, maximizing its cotransporter activity [15].

A lack of stem cell assays

A lack of stem cell assays in the adult salivary gland field has hindered studies aimed at assessing functional properties and/or regenerative potential of putative stem cell populations. Our previous attempts for studying SGSC biology were based on in vitro cultures, already enriched for stem/progenitor cells (Lombaert et al., 2008a; Nanduri et al., 2014). Here we report the development of an optimized culture system for generating salivary gland organoids from primary adult submandibular glands in vitro, based on activation of Wnt signaling. Although in addition to the common transmembrane expression of EpCAM we also detect an unexpected basal localization of the protein in excretory duct cells of the salivary gland, FACS selection for single EpCAMhigh cells and subsequent culturing in the presence of Wnt-activating proteins efficiently generate three-dimensional salivary gland organoids that closely resemble primary salivary gland tissue.
Recently, Xiao et al. (2014) successfully isolated and cultured Lin−CD24+c-Kit+Sca1+ SGSCs as spheres lacking the phenotypical hallmark of differentiation: branching morphogenesis. Contrary to this, the organoids presented here underwent a series of SCR 7 events, until reaching sizes up to 1 mm (Figure S2C). Combined with orthotopic transplantation (Ogawa et al., 2013), these findings may open up novel routes to organ-replacement regenerative therapy. Using the technique reported here we are able to expand SGSCs, derived from a single animal, to clinically relevant numbers without the use of specific stem cell markers. Previously reported methods have been required to start expansion cultures with cells derived from multiple animals, and used cell-surface markers that are mouse specific (Xiao et al., 2014), rendering the expansion protocol less clinically relevant.
Achieving control of cell fate determination in adult tissues is one of the key goals of regenerative medicine. Given the central role of Wnt-mediated cellular responses in stem cell self-renewal, we focused our attention on activation as well as inhibition of Wnt signaling. By using a panel of chemical inhibitors of the Wnt pathway, we effectively show that Wnt signaling is required for the maintenance of SGSC cultures. Although the ability of IWR and IWP compounds to selectively inhibit the Wnt pathway has been characterized elsewhere (Chen et al., 2009), it is still possible that chemical inhibition causes off-target effects. Therefore, in the future it would be of interest to use a conditional β-catenin loss-of-function mouse model (Huelsken et al., 2001) or the CRISPR-Cpf1 genome editing system (Zetsche et al., 2015) for the ablation of Wnt pathway.
Ultimately, this study provides proof of principle that SGSCs cultured under Wnt-inducing conditions can be used for stem cell therapy to irradiation-damaged epithelium and possibly other cases of salivary gland dysfunction. Remarkably, the transplanted cells adhere and engraft into damaged tissue and contribute to the normal homeostasis of the salivary gland. We report an unprecedented improvement of saliva flow recovery over previously reported methods (Lombaert et al., 2008b; Nanduri et al., 2011, 2014; Xiao et al., 2014), which could have been achieved by transplantation of a heterogeneous pool of cells containing stem, progenitor, and differentiated cells present in these cultures. Although translation to the human situation is needed, the current study implies that in vitro expansion and transplantation of long-term cultured SGSCs may be a promising option for patients with severe salivary gland hypofunction.
In conclusion, we provide clear evidence that Wnt signals are necessary for SGSC maintenance in vitro. However, this does not exclude the relevance of other pathways in these processes. For example, platelet-derived growth factor receptor signaling, in concert with fibroblast growth factor signaling, has been shown to be essential for proliferation and survival of ex vivo cultures of embryonic salivary gland progenitors (Steinberg et al., 2005; Yamamoto et al., 2008). Furthermore, EGFs and their receptors are important for embryonic salivary gland proliferation (Haara et al., 2009; Knox et al., 2010). However, none of these pathways have been exploited to study adult stem cell maintenance in vitro to the extent as the Wnt-signaling pathway presented here. We believe that the efficient in vitro system reported here will be of vital use for validating and implementing further studies on adult SGSC biology and for the discovery of novel pathways involved in the maintenance and regeneration of salivary glands.

The ubiquitously expressed FGF is the prototypic

The ubiquitously expressed FGF2 is the prototypic member of the paracrine group of human FGFs (Beenken and Mohammadi, 2009). Its endogenous expression is regulated at multiple levels including the alternative translation of polypeptides that eventually differ in size (18–34kDa), domain structure and intracellular localization (Yu et al., 2007). Although experimental results suggest the function of FGF2 in angiogenesis and the regulation of the vascular tone, its physiological roles remain unclear as FGF1−/− and FGF2−/− double knock-out mice are completely viable and fertile showing only minor defects in SCR 7 development, haematopoiesis and wound healing (Cuevas et al., 1991; Fulgham et al., 1999; Miller et al., 2000; Zhou et al., 1998). FGFs signal via structurally related cell surface receptors (FGFR1-5) that, apart from FGFR5, function as receptor tyrosine kinases (Mohammadi et al., 2005; Sleeman et al., 2001). The multiple isoforms of FGFR1-3 generated by exon skipping and alternative splicing have different ligand specificity and tissue-specific expression (Eisemann et al., 1991; Johnson et al., 1991). The signalling machinery of FGFs is similarly complex and, based on literature data, linked to the PLCγ1, the PKB/Akt and the ERK pathways alike (Chen et al., 2000; Chikazu et al., 2000; Park et al., 1999). The effects of the 18kDa species of FGF2 used on hBMSC include the preservation of their fibroblast morphology as well as proliferation and differentiation capacity (Martin et al., 1997). Interestingly, these effects are similar to those of hypoxia on FGF-naïve hBMSCs including the enhanced proliferation, prevented senescence, up-regulated metabolism and, apart from the osteogenic lineages, increased differentiation efficiency (Grayson et al., 2007; Kanichai et al., 2008; Tsai et al., 2011; Volkmer et al., 2010).
The primary intracellular sensors of hypoxia are the prolyl hydroxylase enzymes (PHDs) that, utilising oxygen as co-factor, hydroxylate critical proline residues of their substrates (Willam et al., 2004). PHDs target the basic helix-loop-helix transcription factor hypoxia inducible factor-1 (HIF-1) (Wang et al., 1995). Active HIF-1 consists of one alpha and one beta subunits that are constitutively translated in most cells types (Wang and Semenza, 1995). PHD-mediated prolyl hydroxylation of the alpha subunit, however, induces its ubiquitination and proteasomal degradation preventing the formation of the active HIF-1α/β heterodimers (Chan et al., 2002; Ivan et al., 2002; Masson et al., 2001). In contrast, under hypoxic conditions, PHDs become inactive leading to the accumulation of the functional HIF-1 heterodimers. Once stabilised, active HIF-1 alters the transcriptional pattern to facilitate cellular adaptation to the hypoxic milieu (Majmundar et al., 2010).
Although the molecular machinery orchestrating the biological responses to hypoxia at the transcriptional level is well detailed, the relationship between the FGF2 and hypoxia signalling in hBMSCs has remained unexplored. The similar effects of FGF2 and hypoxia on hBMSC cultures, however, suggest that the two stimuli utilise, at least in part, the same molecular machinery. This raises the question if the use of FGF2 upon the in vitro expansion of hBMSCs has an impact on the hypoxic adaptation of the same. We investigated this question by combining methodologies of molecular biology and biophysics. Here, we provide data that FGF2 enhances the expression of genes critical in the metabolic adaptation to hypoxia. We found that both FGF2 and hypoxia signal through the ERK pathway and the two signals are merged upstream of the mitogen activated protein kinase kinase (MEK). The simultaneous activation of the ERK pathway in FGF2-cultured hypoxic hBMSCs leads to altered signalling dynamics and enhanced HIF-1α DNA binding activity. We found that the latter effect is mediated post-translationally by direct interaction of the activated ERK and stabilised HIF-1α proteins. We also provide data that FGF2 not only facilitates the reduction of the nuclear mobility of HIF-1α but also results in modified complex formation in hypoxic cells.

J Baillargeon RJ Urban YF Kuo HM Holmes

. J Baillargeon, RJ Urban, YF Kuo, HM Holmes, MA Raji, A Morgentaler, BT Howrey, YL Lin, KJOttenbacher. Public Health Rep 2015;130:143–52
Testosterone therapy for late-onset hypogonadism is a controversial practice. What is not controversial is the need for appropriate patient selection and careful monitoring for treatment emergency adverse events in those patients who do choose to initiate SCR 7 therapy. While individual practitioners may vary in the details of their follow-up protocols, all experts agree that patients should be followed. This follow-up must include physical examination and routine laboratory testing.

Introduction
Squamous cell carcinoma of the penis (PC) is a rare cancer with an incidence of <1 in 100,000 men in North America and Europe [1]. Local control of invasive PC has traditionally been achieved with partial penectomy with a 2-cm margin, but recent literature shows more conservative resection margins have no effect on long-term oncologic control with the potential to further preserve acceptable sexual and urinary function [2-4]. While various studies have reported on functional outcomes after organ-preserving surgeries including glansectomy (removal of the glans) or distal corporectomy (removal of glans and <2-cm margin of distal corpora cavernosa), few have utilized a validated questionnaire to assess postoperative outcomes. Our objective was to investigate patient satisfaction with their sexual and urinary outcomes following organ-preserving surgery, including glansectomy or distal corporectomy, for carcinoma of the penis.
Patients

Methods
Approval was obtained by the Cleveland Clinic Institutional Review Board, and consent was obtained via telephone. Patients were asked to complete two questionnaires addressing their health and quality of life (QOL) and return the results by mail. The International Index of Erectile Function (IIEF-15) was utilized as a validated questionnaire to assess erectile function and satisfaction with sexual activity, and results were used to calculate a specific score for each patient regarding each of the four domains of sexual function represented on the IIEF [5].
The patient-reported outcome measure (PROM) for urethral stricture surgery, as reported by Jackson et al., was utilized as a validated questionnaire to quantify changes in voiding symptoms and health-related QOL following penile surgery [6]. The survey included 22 questions addressing urinary symptoms, sexual function, and perceptions of overall health and satisfaction with the operation.
Sociodemographic data and operation details were obtained through chart review and were codified and arranged in tables with questionnaire results. Patients were provided with a $25 Amazon.com gift card as compensation.

Results

Discussion
Penile cancer is most common in Asian, African, and South American countries where it accounts for up to 10% of cancers in men [1]. Poor hygiene is a risk factor most commonly associated with penile carcinoma, and high rates of early male circumcision likely explain low rates of PC in the United States. The infrequency with which PC is seen in Western, circumcised males has made it difficult to acquire data for studies addressing basic questions patients may have regarding postoperative sexual and urinary outcomes. Our study is the first to use standardized, validated questionnaires to evaluate sexual and urinary function in a U.S. patient population [7]. Patients were overall receptive to both the IIEF and PROM for urethral stricture questionnaires, with six patients matching study criteria consenting to the study and completing both questionnaires.
Maintenance of adequate sexual function is a very common concern for men undergoing treatment for penile cancer. It has been previously shown that seven of 25 men treated for penile cancer by various methods reported that, if asked again, they would choose a treatment with lower long-term survival to increase the chance of remaining sexually potent [8].

To see the effects of and on

To see the effects of and on temperature distribution, we have plotted Fig. 3. It is observed that the temperature profiles are linear for lower values of the parameters and while it becomes parabolic for higher values. Further, temperature is gradually enhanced with increasing the values of the parameters. The influence of the parameters of and on concentration distribution is displayed in Fig. 4. Fig. 4a is plotted for various values of chemical reaction parameter . It shows that increasing enhances the fluid concentration. The opposite trend is seen for the case when is increased as noted in Fig. 4b. To see the influence of the parameters and on coefficient of heat transfer (Z), we have displayed in Fig. 5. It shows the oscillatory behavior of heat transfer which may be due to peristalsis. It depicts that maximum amplitude of Z enhanced with increasing , and while the opposite is true for increasing .
The formation of an internally circulating SCR 7 of fluid by closed streamlines is shown in Fig. 6. This trapped bolus pushed a head along peristaltic waves. The aim of Fig. 6 was to examine the influence of different parameters on trapping. Fig. 6a shows that the trapped bolus decreases with increasing elastic parameter . The influence of on trapping is displayed in Fig. 6b. It depicts that streamlines are increased with an increase of . The quite opposite effect can be noticed when increasing the parameter , which is shown in Fig. 6c. Fig. 6d shows the opposite effect of Fig. 6b, if is replaced by magnetic parameter. Furthermore, our results are in good agreement with the results of Eldabe et al. [25] (for non-porous case) by choosing (see Fig. 7).

Conclusion
In this section, numerical calculations have been performed in order to see the dependence of heat and mass transfer characteristics of MHD peristaltic transport of a dusty fluid through a horizontal channel for both fluid and solid particles. Analytical solutions for the problem are obtained by using perturbation technique for both fluid and solid particles. The features of flow, heat and mass transfer characteristics are presented and discussed graphically. The effects of pertinent parameters on flow, heat and mass transfer characteristics have been studied. Investigation of such analysis is of utmost importance owing to its wide range of applications in engineering and biological systems. In particular, it may serve for the intrauterine fluid motion in a sagittal cross section of the uterus under cancer therapy and drug analysis, the transport of lymph in the lymphatic vessels, and the vasomotion in small blood vessels such as arterioles, venues, and capillaries [1–14]. The main observations of the presented attempt may be summarized as follows:

Introduction
The Lotka–Volterra model describes an arbitrary number of ecological competitors (or predator–prey) model which is dynamic by nature [1]. This model, based on the ecological system was framed and gradually gained its popularity in the technological arena. The simple prey–predator model is among the most popular models, being frequently used to demonstrate a simple non-linear control system.
In the concerned field of science and technology, numerous significant physical phenomenons are frequently modeled by nonlinear differential equations. Such equations are often stiff or impractical to solve analytically. Yet, analytical approximate methods to obtain fairly accurate solutions have gained much significance in recent years [18]. There are numerous methods, undertaken to find out approximate solutions to nonlinear problems: Homotopy Perturbation method (HPM), Homotopy Analysis method (HAM) [21], Differential Transform method (DTM) [15–17], Variational Iteration method (VIM) [22], Adomian Decomposition method (ADM), Laplace Adomian Decomposition method (LADM) and Runge–Kutta–Fehlberg method (RKF) and Chebyshev Spectral methods [19,20] are some proven instances. The purpose of this paper was to bring out the analytical expressions of Lotka–Volterra prey predator model and the solution of nonlinear differential equations by using the new approach to Runge–Kutta–Fehlberg method (RKF) in an elegant way. Thus all these methods entail to multidimensional aspects.

After the TEM in situ heat treatment measurements had been

After the TEM in situ heat treatment measurements had been carried out the sample chip with the film was studied using SEM, see Fig. 10. It was found that the film was covered with large micrometer sized PCBM crystals and that there is a variation in size depending on the distance of the crystal from the center of the platinum spiral used to heat the sample. This indicates that there is a temperature gradient over the area, with a higher temperature in the middle. We have previously investigated the connection between nucleation and growth rate of PCBM crystals, and annealing temperature and time in TQ1:PCBM films [17]. in these investigations we found that PCBM crystals grow faster at higher temperatures. That crystals cover the film is not surprising. Only small regions of the film were irradiated, where a majority of the regions where located on the silicon nitride windows that can be found on the sample grid, see Fig. 10. Most of the film was thus not exposed to the electron beam and nothing stopped crystals from starting to nucleate and grow when the film was heated in-situ. There are however two more regions of the film that were irradiated during the in-situ measurements. These are regions of the film that were exposed to the electron beam at the start of the in-situ measurements, when the valve between the column and the electron gun of the microscope was opened. These regions can be seen as two circular areas where no crystals have formed during the in-situ heating of the film. The short exposure these areas received to the electron beam is enough to completely inhibit the nucleation and growth of PCBM crystals, once again verifying that electron irradiation quenches the nucleation of PCBM crystals.

Conclusion
We have used TEM, SEM and UV–vis spectroscopy to investigate how electron irradiation can be used to increase the thermal stability of photovoltaic blends. The results of our investigations show that irradiation with low-energy electrons enhances the thermal stability of TQ1:PCBM films to such a degree that the nanostructure of a film will remain stable even under severe heating conditions, i.e. substantially above the Tg of the blend. Our results also show that the electron irradiation does not seem to noticeably degrade the UV–vis SCR 7 of a TQ1:PCBM film. Thus, electron irradiation can potentially serve as a useful and cost-efficient tool to increase the thermal stability of OSC, possibly even as a tool to optimize the PCE of a device. Heat treatment can be used to establish an optimized nanostructure and electron irradiation can then be used to lock the optimized nanostructure.

Acknowledgements
We thank the Swedish Research Council (Grant no. D0565301), the Swedish Energy Agency (Grant no. 36643-1), the Chalmers Area of Advance Materials Science, the Chalmers Area of Advance Nanoscience and Nanotechnology and the Knut and Alice Wallenberg Foundation (Grant no. 2012.0339) for financial support.

Introduction
Developments in graphene research have stimulated intense interest in the study of other two-dimensional (2-D) materials such as boron nitride (BN) and MoS2[1–5]. These have considerable application prospects, and overcome some of graphene\’s limitations. For example hexagonal boron nitride (hBN) is structurally similar to graphene but has complementary properties. It is a good insulator with a bandgap of ∼5.9eV [6] and in its 2-D form has proven itself as prominent insulator for graphene-based electronic devices [7,8]. 2-D materials show very promising prospects for applications in opto-electronics, for which the possibility of bandstructure tailoring is vital. This is a particularly essential requirement for graphene, which does not possess a bandgap [9,10]. Here, electronic doping other than via electric gauging or chemical functionalisation would be a major breakthrough. This applies to the functionalisation of all 2-Ds, since chemical methods have significant drawbacks due to lack of control, contamination and resulting secondary impurities, as well as instability and site selectivity. Although successful n-doping with N via the chemical route and subsequent annealing in the case of graphene has been reported [11], introducing dopants in a controlled, pristine fashion via low-energy ion implantation, as theoretical investigations have been suggesting [12], is highly desirable. By enabling flexible small-depth channel doping, ion implantation has revolutionised silicon and semiconductor-technology resulting in a significant economic and societal impact. Integration of 2-Ds into these technologies will be a further and major step-up. If ion implantation for electronic doping was proven to be successful, industrial implantation facilities could be re-fitted with low energy implantation sources. Thus, by controlling ion beam energies one could envisage fabrication of spatially non-uniform 2-D integrated materials. P- and n-doped areas could then be designed with a resolution greater than 5nm, enabling the creation of nanoscale heterostructures, such as lateral quantum dots. This presents significant prospects for industrial scale non-chemistry-reliant functionalisation and processing of 2D materials.

Yellow white laccases are rarely studied unlike blue

Yellow/white laccases are rarely studied unlike blue laccases. The major difference between yellow and blue laccases is the lack of an SCR 7 band at 610nm always found in blue laccases. As a matter of fact, yellow laccases are known to catalyze oxidation without the need for mediators and this makes yellow laccases a better biocatalyst than blue laccases [13]. In this study, Aureobasidium pullulans, a black-yeast-like fungus, of immense biotechnological application (such as the production of a battery of industrially important enzymes [14], polysaccharide (pullulans) and antimycotic agent, aureobasidin A [15]) was isolated from soil containing decayed plant litters at an unfarmed site (Latitude N 7°31.2006′ and Longitude E 4°31.5797′), in the Department of Botany, Obafemi Awolowo University Campus, Ile-Ife, Nigeria. Since yellow laccases are often less studied with more focus on the blue laccases, this study investigated the yellow laccase elaborated by A. pullulans NAC8, which was subsequently purified, biochemically characterized and the catalytic properties determined. Preliminary investigations on the utilization of this enzyme in decolorization of textile dyes and textile waste water effluents have been carried out in our laboratory [16]. The catalytic properties and laccase type of this enzyme from A. pullulans NAC8, has not been reported in any literature. The possible biotechnological applications of this yellow laccase such as in biocatalysis and possible utilization in the detoxification of textile dyes makes it necessary to explore its biochemical and catalytic characteristics.

Materials and methods

Results and discussion
The fungal strain, A. pullulans NAC8, shared 82% homology with A. pullulans HN2.3, 79% homology with Aureobasidium mansonii strain ATCC36276 and 100% homology with A. pullulans strains P-18, YY7. The phylogenetic tree was drawn using NJPLOT after alignment of the sequences with the Clustal X software (Fig. 1). The sequence was deposited in the gene bank of the NCBI (Accession No: KX023301). The choice of amplifying the 5.8S gene and the adjacent ITS regions 1 and 2 was to determine the phylogenetic position of the strains. ITS regions are highly variable and can only be aligned with confidence when comparing closely related taxa. Recently, laccase production by several strains of A. pullulans was carried out by Rich et al. [27].
Laccase from A. pullulans was purified using ion-exchange chromatography on DEAE-Sephadex and dialysis of the active pooled fractions against glycerol had a single peak (Fig. 1) of activity was obtained a final yield of 59.3% and a purification fold of 2.0. The summary of a typical purification is shown in Table 1. The enzyme bound to the ion exchanger and hence it is anionic. Laccase from Gaeumannomyces graminis had a 4.6% yield and 120-fold purification [21] using a combination of dialysis and ultrafiltration. Laccase from Bacillus sp. ADR was purified using DEAE-anion exchanger with 33% yield and 56 purification fold [22]. A distinct band was obtained corresponding to 68.4kDa (Fig. 2) hence the molecular weight determined on SDS–PAGE was 68.4kDa. Fungal laccases vary in their molecular weight but most fall within the range of 60–70kDa [23]. The molecular weight reported for A. pullulans laccase varies. Rich et al. [27] reported molecular weights above 70kDa after deglycosylation with Endo H and even a particular strain, NRRYL-2568 had a molecular weight of 100kDa after treatment with Endo H. This suggests that the degree of glycosylation affects the molecular weight from several strains of A. pullulans.
The concentrated enzyme was yellow in color, the absorption from 250 to 800nm gave just a single peak at 280nm (Fig. 4). There was no peak shown at 610nm. The ratio of absorption at 280nm (A280) to absorption at 610, (A610) was 26. This is characteristic of yellow laccases. The ratio of absorption at 280nm (A280) to absorption at 610, (A610) was determined to be 26.0. This was higher than (15–20) meant for blue copper laccases. A value of 36.0 was obtained for Ganoderma fornicatum laccase [24]. This suggests that A. pullulans NAC8 laccase is a yellow laccase rather than a blue one. Yellow laccases are able to catalyze the oxidation of non-phenolic aromatic compounds in the absence of exogenous mediators [25]. The atomic absorption spectrophotometry showed that the ratio of copper to manganese is 3:1. Laccases contain 4 copper atoms termed Cu T1 (where the reducing substrate binds) and trinuclear copper cluster T2/T3 (where oxygen binds and is reduced to water). The three copper atoms can be distinguished using UV/visible SCR 7 and electronic paramagnetic resonance (EPR) spectroscopy [1]. There have been reports of yellow laccases having less than four copper atoms with the copper atoms replaced with manganese or other metals. Telke et al. [22] reported the existence of 2 copper atoms in Bacillus sp. ADR laccase. Atomic absorption spectrophotometry indicated that laccase from A. pullulans has three copper atoms and one magnesium atom. Palmeiri et al. [26] reported the existence of just a copper atom instead of four in Pleurotus ostreatus. Most non blue laccases probably have the fourth copper atom replaced with another metal and it might be responsible for lack of absorption at 610nm. (See Fig. 4).

The purpose of this study was to evaluate diaphragmatic motion

The purpose of this study was to evaluate diaphragmatic motion during tidal breathing in a standing position in a health screening center cohort using dynamic chest radiography in association with participants\’ demographic characteristics.

Materials and Methods

Study Population

This cross-sectional study was approved by the institutional review board, and all the participants provided written informed consent. From May 2013 to February 2014, consecutive 220 individuals who visited the health screening of our hospital and met the following inclusion criteria for the study were recruited: age greater than 20 years, scheduled for conventional chest radiography, and underwent pulmonary function test. Patients with any of the following criteria were excluded: pregnant (n  =  0), potentially pregnant or lactating (n  =  0), refused to provide informed consent (n  =  22), had incomplete datasets of dynamic chest radiography (n  =  3), had incomplete datasets of pulmonary function tests (n  =  1), could not follow tidal breathing instructions (eg, holding breath or taking a deep breath) (n  =  18), or their diaphragmatic motion could not be analyzed by the software described next (n  =  4). Thus, a total of 172 participants (103 men, 69 women; mean age 56.3 ± 9.8 years; age range 36–85 years) were finally included in the analysis ( Fig 1). The data from 47 participants of this study SCR 7 were analyzed in a different study (under review). The heights and weights of the participants were measured, and the body mass index (BMI, weight in kilograms divided by height squared in meters) was calculated.

Figure 1. Flow diagram of the study population.Figure optionsDownload full-size imageDownload high-quality image (83 K)Download as PowerPoint slide

Imaging Protocol of Dynamic Chest Radiology (“Dynamic X-Ray Phrenicography”)

Posteroanterior dynamic chest radiography (“dynamic X-ray phrenicography”) was performed using a prototype system (Konica Minolta, Inc., Tokyo, Japan) composed of an FPD (PaxScan 4030CB, Varian Medical Systems, Inc., Salt Lake City, UT, USA) and a pulsed X-ray generator (DHF-155HII with Cineradiography option, Hitachi Medical Corporation, Tokyo, Japan). All participants were scanned in the standing position and instructed to breathe normally in a relaxed way without deep inspiration or expiration (tidal breathing). The exposure conditions were as follows: tube voltage, 100 kV; tube current, 50 mA; pulse duration of pulsed X-ray, 1.6 ms; source-to-image distance, 2 m; additional filter, 0.5 mm Al + 0.1 mm Cu. The additional filter was used to filter out soft X-rays. The exposure time was approximately 10–15 seconds. The pixel size was 388 × 388 µm, the matrix size was 1024  × 768, and the overall image area was 40 × 30 cm. The gray-level range of the images was 16,384 (14 bits), and the signal intensity was proportional to the incident exposure of the X-ray detector. The dynamic image data, captured at 15 frames/s, were synchronized with the pulsed X-ray. The pulsed X-ray prevented excessive radiation exposure to the subjects. The entrance surface dose was approximately 0.3–0.5 mGy.

Image Analysis

The diaphragmatic motions on sequential chest radiographs (dynamic image data) during tidal breathing were analyzed using prototype software (Konica Minolta, Inc.) installed in an independent workstation (Operating system: Windows 7 Pro SP1; Microsoft, Redmond WA; CPU: Intel Core i5-5200U, 2.20 GHz; memory 16 GB). The edges of the diaphragms on each dynamic chest radiograph were automatically determined by means of edge detection using a Prewitt Filter 18 ;  19. A board-certified radiologist with 14 years of experience in interpreting chest radiography selected the highest point of each diaphragm as the point of interest on the radiograph of the resting end-expiratory position (Fig 2a). These points were automatically traced by the template-matching technique throughout the respiratory phase (Fig 2b, Supplementary Video S1), and the vertical excursions of the bilateral diaphragm were calculated (Fig 2c): the null point was set at the end of the expiratory phase, that is, the lowest point (0 mm) of the excursion on the graph is the highest point of each diaphragm at the resting end-expiratory position. Then the peak motion speed of each diaphragm was calculated during inspiration and expiration by the differential method (Fig 2c). If several respiratory cycles were involved in the 10 to 15-second examination time, the averages of the measurements were calculated.

In the tumor group lactate alone was

In the tumor group, lactate alone was found in 12 patients, lactate and choline were seen in 5 patients, neither lactate nor choline in 2 patients, lipid and lactate seen in 2 metastatic brain patients, none of the patients showed aminoacids, succinate or acetate. NAA and creatine were reduced or absent in all patients.
With DWI, 9 cases of abscess were correctly diagnosed and 2 cases of abscess were misdiagnosed as tumor, 18 cases of the tumor group were correctly diagnosed and 1 case of the tumor group was misdiagnosed as an abscess.
With MRS two abscesses were misdiagnosed as tumor, these findings may due to the absence of SCR 7 because these patients were undergoing treatment with antibiotics and all 19 patients in the tumor group were correctly diagnosed.
Table 4.
Comparison between DWI and 1HMRS in differentiating between brain abscess and necrotic tumors.Sensitivity (%)Specificity (%)PPV (%)NPV (%)DWI94.7381.8290901HMRS1008290100DWI = diffusion-weight imaging, 1HMRS = proton magnetic resonance spectroscopy.Full-size tableTable optionsView in workspaceDownload as CSV
Figs. Fig. 1, Fig. 2, Fig. 3 and Fig. 4 demonstrate a sample of selected cases of our study, each figure outline one case.
Fig. 1. (A-E): A male patient aged 61 years shows ring enhancing lesion in the right deep parietal region giving the radiological appearance of brain abscess but by clinical follow up and with antibiotics, no improvement could be detected and in biopsy, low grade astrocytoma was diagnosed. (A) and (B) Axial pre and post contrast T1WI show well defined ring enhancing brain lesion in the right deep parietal region. (C) Diffusion weighted Images show: The central portion of the lesion displays high signal intensity. (D) ADC shows: The central cavity of the lesion displays low signal (black arrow) (E) 1HMRS shows: The central portion shows resonance peaks of, lactate (Lact) and lipid (Lip) but no peaks of amino acids which is the signature of abscess.Figure optionsDownload full-size imageDownload as PowerPoint slide

For all commodities the percentage of

For all commodities, the percentage of observations in the low regime of volatility ranges from 70.4% to 86.9%. Except for corn, the threshold values chosen by the algorithm are homogeneous between price pairs of each commodity.
4.4.1.2. Price adjustments to the long term relationship
On 48 coefficients of long term adjustments αi,s, 27 are significant at a minimum level of 10%. In all the cases, the coefficients have the correct signs. Indeed, for each pair, the coefficient of long term adjustment of the reference price αA,s is negative and that of the producer price is positive. Hence, when the price difference of PA ? PB increases, namely PA increases and/or PB decreases, the error correction mechanism leads to a SCR 7 in PA and an increase in PB and vice versa because in the long run the price difference reverts back to the amount of transfer costs.
The analysis of results begins by commodities with a futures market, namely rapeseed and corn.
For rapeseed price pairs, in both volatility regimes, producer prices are the only ones that adjust to the long term relationship. Nonetheless, even if there is no significant nonlinearity the adjustment is faster in the high regime of volatility. The coefficients αB,1 range from 0.317 to 0.408 in the low regime against values between 0.608 and 0.797 for the coefficients αB,2 in the high regime. In term of half-life, the effect of a shock on the long term relationship is equal to 0.93-1.24 week7 in the first regime against 0.53-0.79 week in the second regime. In other words, the adjustment process is approximatively two times faster in the second regime of volatility.
For the two corn price pairs, in the inferior regime of volatility, only producer prices adjust to deviations from the long term equilibrium. For the Bordeaux-Nord-Toulouse pair, both react to the long term relationship. Adjustment coefficients for producer prices are from 0.137 to 0.166. In the high volatility regime, the dynamics remains the same but as for rapeseed coefficients αi,2 are greater in absolute values. For example, for the Bordeaux-Puy-de-Dôme pair, the value of αB,s increases from 0.150 to 0.279 which is equivalent to a change in the half-life of a shock from around 7 to 2.5 weeks. Great reductions are also observed for the two other pairs.
The estimations of TVECM with GARCH volatility for feed barley are reported in Table 6. In the inferior regime of volatility, for two cases over three, both αA,1 and αB,1 coefficients are significant at 10%. This implies that the reference price (Rouen) and producer prices both respond to deviations from the long term equilibrium. It appears that αA,1 are greater than αB,1. In the Rouen-Eure-et-Loir pair, only the reference price reacts to the long term equilibrium with a αA,1 coefficient equals to ? 0.129. In the superior regime of volatility, results strongly contrast from rapeseed and corn. Only the reference price adjusts to the long term equilibrium. The coefficients αA,2 ranges from ? 0.262 to ? 0.436. The Rouen-Eure-et-Loir pair has the highest coefficient which is equivalent to a half-life of 2.54 weeks against 5.37 in the first regime of volatility. For the two other pairs, half-lives are 1.29 and 1.75 in the high volatility regime against 2.24 and 2.14 in the low volatility regime.