Filler Mixed Into Adhesives Does Not Necessarily Improve Their Mechanical Properties.

OBJECTIVES: To investigate the influence of filler type/loading on the micro-tensile fracture strength (µTFS) of adhesive resins, as measured 'immediately' upon preparation and after 1-week water storage ('water-stored'). METHODS: The morphology and particle-size distribution of three filler particles, referred to as 'Glass-S' (Esschem Europe), 'BioUnion' (GC), and 'CPC_Mont', were correlatively characterized by SEM, TEM, and particle-size analysis. These filler particles were incorporated into an unfilled adhesive resin ('BZF-29unfilled', GC) in different concentrations to measure the 'immediate' µTFS. After 1-week water storage, the 'water-stored' µTFS of the experimental particle-filled adhesive resins with the most optimum filler loading, specific for each filler type, was measured. In addition, the immediate and water-stored µTFS of the adhesive resins of three experimental two-step universal adhesives based on the same resin matrix but varying for filler type/loading, coded as 'BZF-21' (containing silica and bioglass), 'BZF-29' (containing solely silica), and 'BZF-29_hv' (highly viscous with a higher silica loading than BZF-29), and of the adhesive resins of the gold-standard adhesives OptiBond FL ('Opti-FL', Kerr) and Clearfil SE Bond 2 ('C-SE2', Kuraray Noritake) was measured along with that of BZF-29unfilled (GC) serving as control/reference. Statistics involved one-way and two-way ANOVA followed by post-hoc multiple comparisons (α<0.05). RESULTS: Glass-S, BioUnion, and CPC_Mont represent irregular fillers with an average particle size of 8.5-9.9 µm. Adding filler to BZF-29unfilled decreased µTFS regardless of filler type/loading. One-week water storage reduced µTFS of all adhesive resins except BZF-21, with the largest reduction in µTFS recorded for BZF-29unfilled. Among the three filler types, the µTFS of the 30 wt% Glass-S and 20 wt% BioUnion filled adhesive resin was not significantly different from the µTFS of BZF-29unfilled upon water storage. CONCLUSIONS: Adding filler particles into adhesive resin did not enhance its micro-tensile fracture strength but appeared to render it less sensitive to water storage as compared to the unfilled adhesive resin investigated.

Acrylamide monomers in universal adhesives.

OBJECTIVES: The mono-functional monomer 2-hydroxyethyl methacrylate (HEMA) is often added to universal adhesives (UAs) to improve surface wetting and prevent phase separation. Nevertheless, HEMA promotes water sorption and hydrolysis at adhesive interfaces, hereby affecting long-term bonding to dentin. This study investigated if two acrylamide monomers could replace HEMA in an UA formulation applied in etch-and-rinse (2E&R) and self-etch (1SE) bonding mode. METHODS: Four experimental UAs were bonded to bur-cut dentin. In addition to 12 wt% 10-MDP, 25 wt% Bis-GMA and 10 wt% TEGDMA as common monomer composition, 20 %wt ethanol and 15 %wt water as solvent, and 3 wt% polymerization-related additives, the four formulations solely differed for either the acrylamide cross-linker monomer 'FAM-201' as TEGDMA alternative and HEMA replacement, the hydroxyethyl acrylamide monomer 'HEAA' as HEMA alternative, HEMA ('HEMA+'), or extra TEGDMA in a HEMA-free control ('HEMA-'), all added in a 15 wt% concentration. The split-tooth study design involved application in 2E&R mode on one tooth half versus 1SE mode on the corresponding half. Micro-tensile bond strength of half of the micro-specimens was measured upon 1-week distilled water storage ('immediate' 1w µTBS), with the other half measured after additional 6-month storage ('aged' 6 m µTBS). Statistics involved linear mixed-effects (LME) modelling (p < .05). Additionally, interfacial TEM characterization, thin-film (TF) XRD surface analysis, LogP determination, and a cytotoxicity assay were carried out. RESULTS: FAM-201 revealed significantly higher µTBS than HEMA+ at 1w and 6 m when applied both in E&R and SE bonding modes. HEAA's µTBS was significantly lower than that of HEMA+ at 1w when applied in SE mode. TF-XRD and TEM revealed similar chemical and ultrastructural interfacial characterization, including stable 10-MDP_Ca salt nano-layering. FAM-201 was least cytotoxic and presented with an intermediary LogP, while HEAA presented with the highest LogP, indicating high hydrophilicity and water-sorption sensitivity. SIGNIFICANCE: The acrylamide co-monomer FAM-201 could replace HEMA in an UA formulation, while HEAA not.

Bonding performance of experimental HEMA-free two-step universal adhesives to low C-factor flat dentin.

OBJECTIVES: Experimental two-step universal adhesives (2-UAs) providing a particle-filled hydrophobic adhesive resin with a significant film thickness to hydrophobically seal the adhesive interface were designed and synthesized. This study aimed to characterize their interfacial interaction with dentin, to determine whether the 2-UA formulations achieve durable bonding to low C-factor flat dentin and to measure their water sorption. METHODS: Bonding effectiveness of 2-UAs that combine a 10-MDP-based primer with hydrophobic adhesive resins differing only for filler (BZF-21, BZF-29, and BZF-29_hv) were comparatively investigated with the commercial adhesive Clearfil SE Bond 2 (C-SE2, Kuraray Noritake). Adhesive-dentin interfaces were characterized with TEM. Adhesive-resin disks were immersed in distilled water at 37 °C for 1 week, 6 months and 1 year to measure water sorption and solubility. 'Immediate' and 'aged' micro-tensile bond strength (µTBS) of the adhesives applied in etch-and-rinse (E&R) and self-etch (SE) bonding mode to low C-factor flat dentin were measured. Statistical analyses involved linear mixed-effects (LME) modelling and Kruskal-Wallis testing (p < 0.05). RESULTS: TEM revealed that E&R hybrid layers were more sensitive to aging than SE hybrid layers. Lower water sorption was recorded for all UAs compared with C-SE2. The immediate µTBS of BZF-21 and BZF-29 was not significantly different from that of C-SE2. The 1-year aged µTBS of all 2-UAs was significantly lower than that of C-SE2, except for BZF-29 applied in E&R mode. A significant reduction in µTBS upon 1-year aging was recorded for BZF-21 and BZF-29 applied in E&R mode. A significant difference in µTBS between E&R and SE bonding modes was recorded for all adhesives except BZF-21. SIGNIFICANCE: Experimental 2-UAs with a hydrophobic adhesive-resin design produced± 20-µm thick adhesive-resin layers, absorbed less water and resulted in bonding performance that was more aging-resistant when applied in SE than in E&R bonding mode. The silica-filled BZF-29 2-UA revealed the most comparable bonding performance with C-SE2 in a low C-factor condition (flat dentin).

Nano-Layering Adds Strength to the Adhesive Interface.

X-ray diffraction (XRD) surface analysis and ultrastructural interfacial characterization using transmission electron microscopy (TEM) confirmed that the functional monomer 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP) self-assembles into nano-layers at adhesive-tooth interfaces. Self-assembled nano-layering is thought to contribute to the durability of bonding to tooth dentin, although this has not been proven yet. In order to disclose this potential bond-durability contribution of nano-layering, we observed the 3-dimensional (3D) spreading of nano-layering by a series of focused-ion-beam (FIB) milled cross sections by scanning electron microscopy (FIB-SEM) and examined the mechanical properties of self-assembled nano-layering using scanning probe microscopy (SPM). A commercial 10-MDP-containing 3-step self-etch adhesive partially demineralized dentin up to submicron depth, forming a submicron hydroxyapatite-rich hybrid layer. TEM chemically and ultrastructurally confirmed the formation of interfacial nano-layering. FIB-SEM 3D reconstructions disclosed a 3D network of self-assembled nano-layering extending from the hybrid layer up to within the adjacent adhesive-resin layer. SPM revealed that nano-layering within the adhesive-resin layer possessed a higher elastic modulus than that of the surrounding adhesive resin, hereby suggesting that nano-layering contributes to the mechanical strength of adhesives like filler particles do. Nano-layering's 3D expanded structure is expected to strengthen the surrounding resin, as well to better interconnect the adhesive-resin layer to the hybrid layer. In conclusion, this exploratory study demonstrated that nano-layering constitutes a strong phase at the adhesive interface, which may contribute to the clinical longevity of the 10-MDP-based bond to dentin.

Lack of Neutralization of 10-MDP Primers by Zirconia May Affect the Degree of Conversion of Dual-cure Resin Cement.

CLINICAL RELEVANCE: Use of zirconia primers with a low pH and a high acidic monomer concentration should be employed in combination with dual-cure resin cements that are less sensitive to an acidic environment. Primers with lower 10-MDP concentrations attain better outcomes.

Touch-Cure Polymerization at the Composite Cement-Dentin Interface.

Ceramic restorations are often adhesively luted onto the tooth prep. The so-called touch-cure concept was developed to yield optimum polymerization of composite cement at the restoration-cement-tooth interface for immediate bond stabilization. Although this touch cure is theorized to initiate polymerization at the interface when the accelerator in the primer makes contact with the cement, this process has not yet been proven. This study aimed to elucidate the mechanism of touch cure by measuring the degree of conversion (DC) of composite cement applied with or without an accelerator-containing tooth primer (TP) versus an accelerator-free primer using real-time Fourier-transform infrared spectroscopy (RT-FTIR) and attenuated total reflection (ATR)-FTIR. Interfacial bond strength was measured in shear mode, the accelerator composition confirmed by X-ray fluorescence analysis (XRF), and the interfacial interaction of TP and composite cement with dentin investigated by X-ray diffraction (XRD), focused-ion-beam scanning electron microscopy (FIB-SEM) with 3-dimensional interface reconstruction, and transmission electron microscopy (TEM). RT/ATR-FTIR revealed the significantly highest DC when the composite cement was applied with the accelerator-containing primer. XRF disclosed a vanadium compound as a novel chemical accelerator within TP, instead of a classic chemical curing initiator system, to set off touch cure as soon the cement contacts the previously applied primer. Although the TP contains the acidic functional monomer 10-MDP for adhesion to tooth tissue, touch cure using the accelerator-containing TP combined the fastest/highest DC with the highest bond strength. FIB-SEM and TEM confirmed the tight interfacial interaction at dentin with submicron hybridization along with stable 10-MDP also Ca-salt nanolayering.

Multiparameter evaluation of acrylamide HEMA alternative monomers in 2-step adhesives.

OBJECTIVE: As frequently added to adhesives, the mono-functional monomer 2-hydroxyethyl methacrylate (HEMA) acts as co-solvent and improves surface wetting. Nevertheless, HEMA promotes watersorption and hydrolysis at adhesive interfaces, affecting bond durability to dentin. This study investigated if two acrylamide co-monomer alternatives could replace HEMA in experimental adhesive-resin formulations as part of 3/2-step universal adhesives applied, respectively, in etch-and-rinse (E&R) and self-etch (SE) bonding modes. METHODS: Upon priming dentin with the 10-MDP-based Clearfil SE Bond 2' primer ('C-SE2p'; Kuraray Noritake), three experimental adhesive resins, consisting of 50 wt.% Bis-GMA, 15 wt.% TEGDMA, and either 35 wt.% diethyl acrylamide ('DEAA'), hydroxyethyl acrylamide ('HEAA') or HEMA ('HEMA+'), were applied. The control HEMA-free adhesive resin contained 60 wt.% Bis-GMA and 40 wt.% TEGDMA ('HEMA-'). All adhesives were evaluated for 'immediate' and 'aged' micro-tensile bond strength (µTBS) to dentin upon, respectively, 1-week (1w) and 6-month (6m) water storage, TEM adhesive-dentin interfacial interaction, 24-h and 6m three-point bending, contact-angle wetting, viscosity and watersorption. RESULTS: Linear mixed-effects model statistics revealed significantly better bonding performance of the adhesives applied in E&R than SE mode, except for DEAA_1w, with the highest µTBSs recorded for DEAA and HEMA- applied in SE mode. In E&R mode, aging did not significantly reduce DEAA's µTBS. Best wetting on primed dentin was recorded for HEMA+, significantly better than DEAA, further HEAA and HEMA-, these directly related to their viscosity. HEAA absorbed significantly more water than all other adhesive-resin formulations. HEMA->DEAA>HEAA>HEMA+ was the significant order for 6m bending strength. CONCLUSIONS: The acrylamide co-monomer DEAA could replace HEMA, while HEAA not.

Risk-adjusted assessment of incidence and quantity of blood use in acute-care hospitals in Japan: an analysis using administrative data.

BACKGROUND AND OBJECTIVES: Continuous monitoring of blood use and feedback on transfusions are effective in decreasing inappropriate blood transfusions. However, traditional methods of monitoring have practical challenges, such as the limited availability of experts and funding. Administrative data including a patient classification system may be employed for risk-adjusted assessment of hospital-wide blood use. MATERIALS AND METHODS: We conducted an audit of blood use at two hospitals and determined proportions of appropriate blood use at each hospital. We then used administrative data of 587,045 cases provided by 73 hospitals to develop two mathematical models to calculate risk-adjusted use of blood products. The first model is a logistic regression model to predict the percentage of transfused patients. Patient demographics, surgery and diagnostic groups were utilized as predictors of transfusion. The second model is a case-mix adjusted model which predicts hospital-wide use of units of blood products from the distribution of diagnosis-related groups. For each model, the observed to expected (O/E) ratio of blood use in each hospital was calculated. We compared resultant ratios with proportions of appropriate blood use in two of the hospitals studied. RESULTS: Both models showed good prediction abilities. O/E ratios calculated using the two models were relevant to proportions of appropriate transfusions. CONCLUSIONS: Risk-adjusted assessments of blood product use based on administrative data allow hospital-wide evaluation of transfusion use. Comparing blood use between different hospitals contributes toward establishing appropriate transfusion practices.

Distribution and species composition of juvenile and adult scombropids (Teleostei, Scombropidae) in Japanese coastal waters.

Two scombropid fishes, Scombrops boops and Scombrops gilberti, are closely related and commercially important species in Japan. These species are often confused in commercial markets because of their morphological similarity. In this study, scombropid specimens collected from various Japanese coastal waters were subjected to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and phylogenetic analysis of the 16S rRNA gene in mitochondrial DNA. These analyses showed that all the scombropid specimens collected from localities in the Sea of Japan were identified as S. boops, whereas those from the Pacific Ocean included two species, S. boops and S. gilberti. Almost all juvenile (<200 mm standard body length, S(L)) S. gilberti originated from the Pacific coastal waters of the northern Japan, whereas adults (>400 mm S(L)) were found only in deep water off the Izu Peninsula to the Izu Islands. This suggests that S. gilberti might migrate extensively during its life cycle. In addition, differences in the number of specimens and the distribution between the two species suggest that S. gilberti is less abundant than S. boops in Japanese waters.

[Pulmonary venous obstruction after the Williams procedure for partial anomalous pulmonary venous connection].

A 54-year-old man was admitted to our hospital because of chest discomfort. Cardiac catheterization revealed partial anomalous pulmonary venous connection with an intact atrial septum. The right upper pulmonary vein (RUPV) drained into the upper segment of the superior vena cava (SVC). Using the Williams procedure, an atrial septal defect (ASD) was created and a fresh autologous pericardial patch was used to fashion a new pulmonary vein return route from SVC to the ASD. Although the patient was stable after the procedure, he was admitted again 6 months later because of obstruction of RUPV. At reoperation, it was found that the previous pulmonary vein route was obstructed and that the pericardial baffle had adhered to the atrial septum above the ASD. The shrunken and thickened pericardial baffle was removed and the orifice of the ASD was extensively enlarged, after which an expanded polytetrafluoroethylene (ePTFE) patch was used as a new baffle. After the reoperation, the patient's condition improved.

Etching Efficacy of Self-Etching Functional Monomers.

Besides chemically interacting with hard tooth tissue, acidic functional monomers of self-etch adhesives should etch the prepared tooth surface to dissolve the smear layer and to provide surface micro-retention. Although the etching efficacy of functional monomers is commonly determined in terms of pH, the pH of adhesives cannot accurately be measured. Better is to measure the hydroxyapatite (HAp)-dissolving capacity, also considering that functional monomers may form monomer-Ca salts. Here, the etching efficacy of 6 functional monomers (GPDM, phenyl-P, MTEGP, 4-META, 6-MHP and 10-MDP) was investigated. Solutions containing 15 wt% monomer, 45 wt% ethanol, and 40 wt% water were prepared. Initially, we observed enamel surfaces exposed to monomer solution by scanning electron microscopy (SEM). X-ray diffraction (XRD) was employed to detect monomer-Ca salt formation. Phenyl-P exhibited a strong etching effect, while 10-MDP-treated enamel showed substance deposition, which was identified by XRD as 10-MDP-Ca salt. To confirm these SEM/XRD findings, we determined the etching efficacy of functional monomers by measuring both the concentration of Ca released from HAp using inductively coupled plasma-atomic emission spectroscopy (ICP-AES) and the amount of monomer-Ca salt formation using 31P magic-angle spinning (MAS) nuclear magnetic resonance (NMR). ICP-AES revealed that the highest Ca concentration was produced by phenyl-P and the lowest Ca concentration, almost equally, by 4-META and 10-MDP. Only 10-MDP formed 10-MDP-Ca salts, indicating that 10-MDP released more Ca from HAp than was measured by ICP-AES. Part of the released Ca was consumed to form 10-MDP-Ca salts. It is concluded that the repeatedly reported higher bonding effectiveness of 10-MDP-based adhesives must not only be attributed to the more intense chemical bonding of 10-MDP but also to its higher etching potential, a combination the other functional monomers investigated lack.

Predictive population pharmacokinetic/pharmacodynamic model for a novel COX-2 inhibitor.

The objectives of these analyses were to (1) develop a population pharmacokinetic/pharmacodynamic model for a novel COX-2 inhibitor (CS-706) using data from primarily Caucasian subjects, (2) predict responses in subpopulations of interest (including Japanese subjects), and (3) correlate pharmacodynamic parameters to safety outcomes. The model was developed using data from 130 healthy adults following single or multiple doses of CS-706. Serial plasma concentrations of CS-706 and ex vivo whole-blood cyclooxygenase-1 (COX-1) and COX-2 activity were determined up to 72 hours postdose. An E(max) model described relationships between CS-706 plasma concentrations and COX-1 and COX-2 inhibition. CS-706 potency (EC(50)) was 397 ng/mL for COX-1 and 20 ng/mL for COX-2. None of the tested covariates influenced the pharmacodynamics of CS-706. Japanese subjects are expected to show a slightly reduced response to CS-706, consistent with lower exposure following the same dose given to Caucasian subjects. Predictive pharmacokinetic/pharmacodynamic modeling for COX-1 and COX-2 inhibition indicates a 20-fold potency ratio that is expected to be similar in Japanese and Caucasians. There was good correlation between COX-1 inhibition and the incidence of 7-day gastroduodenal mucosal injury. A dose of less than 25 mg bid could be adequate to inhibit COX-2 activity with a low risk of gastrointestinal mucosal injury.

Structural and Chemical Analysis of the Zirconia-Veneering Ceramic Interface.

The interfacial interaction of veneering ceramic with zirconia is still not fully understood. This study aimed to characterize morphologically and chemically the zirconia-veneering ceramic interface. Three zirconia-veneering conditions were investigated: 1) zirconia-veneering ceramic fired on sandblasted zirconia, 2) zirconia-veneering ceramic on as-sintered zirconia, and 3) alumina-veneering ceramic (lower coefficient of thermal expansion [CTE]) on as-sintered zirconia. Polished cross-sectioned ceramic-veneered zirconia specimens were examined using field emission gun scanning electron microscopy (Feg-SEM). In addition, argon-ion thinned zirconia-veneering ceramic interface cross sections were examined using scanning transmission electron microscopy (STEM)-energy dispersive X-ray spectrometry (EDS) at high resolution. Finally, the zirconia-veneering ceramic interface was quantitatively analyzed for tetragonal-to-monoclinic phase transformation and residual stress using micro-Raman spectroscopy (µRaman). Feg-SEM revealed tight interfaces for all 3 veneering conditions. High-resolution transmission electron microscopy (HRTEM) disclosed an approximately 1.0-µm transformed zone at sandblasted zirconia, in which distinct zirconia grains were no longer observable. Straight grain boundaries and angular grain corners were detected up to the interface of zirconia- and alumina-veneering ceramic with as-sintered zirconia. EDS mapping disclosed within the zirconia-veneering ceramic a few nanometers thick calcium/aluminum-rich layer, touching the as-sintered zirconia base, with an equally thick silicon-rich/aluminum-poor layer on top. µRaman revealed t-ZrO2-to-m-ZrO2 phase transformation and residual compressive stress at the sandblasted zirconia surface. The difference in CTE between zirconia- and the alumina-veneering ceramic resulted in residual tensile stress within the zirconia immediately adjacent to its interface with the veneering ceramic. The rather minor chemical elemental shifts recorded in the veneering ceramic did not suffice to draw definitive conclusions regarding potential chemical interaction of the veneering ceramic with zirconia. Sandblasting damaged the zirconia surface and induced phase transformation that also resulted in residual compressive stress. Difference in CTE of zirconia versus that of the veneering ceramic resulted in an unfavorable residual tensile stress at the zirconia-veneering ceramic interface.

Differential expression of apoptotic protease-activating factor-1 and caspase-3 genes and susceptibility to apoptosis during brain development and after traumatic brain injury.

Neuronal apoptosis plays an essential role in early brain development and contributes to secondary neuronal loss after acute brain injury. Recent studies have provided evidence that neuronal susceptibility to apoptosis induced by traumatic or ischemic injury decreases during brain development. However, the molecular mechanisms responsible for this age-dependent phenomenon remain unclear. Here we demonstrate that, during brain maturation, the potential of the intrinsic apoptotic pathway is progressively reduced and that such repression is associated with downregulation of apoptotic protease-activating factor-1 (Apaf-1) and caspase-3 gene expression. A similar decline in apoptotic susceptibility associated with downregulation of Apaf-1 expression as a function of developmental age was also found in cultured primary rat cortical neurons. Injury-induced cytochrome c-specific cleavage of caspase-9 followed by activation of caspase-3 in mature brain correlated with marked increases in Apaf-1 and caspase-3 mRNA and protein expression. These results suggest that differential expression of Apaf-1 and caspase-3 genes may underlie regulation of apoptotic susceptibility during brain development, as well as after acute injury to mature brain, through the intrinsic pathway of caspase activation.

4-Hydroxyretinal, a new visual pigment chromophore found in the bioluminescent squid, Watasenia scintillans.

The bioluminescent squid, Watasenia scintillans has three visual pigments. The major pigment, based on retinal (lambda max 484 nm), is distributed over the whole retina. Another pigment based on 3-dehydroretinal (lambda max approximately 500 nm) and the third pigment (lambda max approximately 470 nm) are localized in the specific area of the ventral retina just receiving the downwelling light. Visual pigment was extracted and purified from the dissected retina. The chromophores were then extracted and analyzed with HPLC, NMR, infrared and mass spectroscopy, being compared with the synthetic 4-hydroxyretinal. A new retinal derivative, 11-cis-4-hydroxyretinal, is identified as the chromophore of the third visual pigment of the squid.

Cloning and characterization of two genes encoding dihydroxyacetone kinase from Schizosaccharomyces pombe IFO 0354.

We report the cloning and characterization of two genes encoding dihydroxyacetone kinase (EC 2.7.1.29), SpDAK1 and SpDAK2, from Schizosaccharomyces pombe IFO 0354. The open reading frames of both genes encode 591 amino acids and have Mrs of 62158 and 62170, respectively. Both predicted amino acid sequences exhibited a high identity to each other (99.8%) and relatively high identities (30% to 76%) to other putative dihydroxyacetone kinase gene products. A Western blot analysis showed that these enzymes are induced by glycerol and repressed by glucose. A genomic Southern blot analysis indicated the presence of SpDAK1 and the absence of SpDAK2 in a standard laboratory strain, S. pombe 972h-.

The effect of recombinant swine interleukin-4 on swine immune cells and on pro-inflammatory cytokine productions in pigs.

The in vitro effect and the in vivo influence of recombinant swine IL-4 (rSwIL-4) were characterized in various swine cells and in nursery pigs on LPS-induced endotoxic shock and pro-inflammatory cytokine productions. In in vitro experiment, the rSwIL-4 induced a proliferation of CD4 positive T cells in mitogen-prestimulated peripheral blood mononuclear cell (PBMC). In addition, the rSwIL-4, which was produced from insect cells, promoted the differentiation of monocytes into immature dendritic cells in combination with granulocyte macrophage-colony stimulating factor (GM-CSF). Furthermore, the rSwIL-4 successfully suppressed the LPS-induced secretion of TNF-alpha, IL-1alpha, IL-6, IL-8, and IL-18 from swine alveolar macrophages when rSwIL-4 was treated at the same time with LPS. In in vivo experiment in nursery pigs, subcutaneous pretreatment of rSwIL-4, which was produced from baculovirus expression system, enhanced the severity of respiratory failure with endotoxic shock, and increased the production of TNF-alpha and IL-18 in response to inoculation with LPS. These results indicate that the rSwIL-4 is biologically active in both in vitro and in vivo treatments. Depending on the administration time, pro-inflammatory cytokine productions by IL-4 can cause either inhibitory or stimulatory regulation.

Comparison of gene expression patterns across 12 tumor types identifies a cancer supercluster characterized by TP53 mutations and cell cycle defects.

Transcriptional profile-based subtypes of cancer are often viewed as identifying different diseases from the same tissue origin. Understanding the mechanisms driving the subtypes may be key in development of novel therapeutics but is challenged by lineage-specific expression signals. Using a t-test statistics approach, we compared gene expression subtypes across 12 tumor types, which identified eight transcriptional superclusters characterized by commonly activated disease pathways and similarities in gene expression. One of the largest superclusters was determined by the upregulation of a proliferation signature, significant enrichment in TP53 mutations, genomic loss of CDKN2A (p16(ARF)), evidence of increased numbers of DNA double strand breaks and high expression of cyclin B1 protein. These correlations suggested that abrogation of the P53-mediated apoptosis response to DNA damage results in activation of cell cycle pathways and represents a common theme in cancer. A second consistent pattern, observed in 9 of 11 solid tumor types, was a subtype related to an activated tumor-associated stroma. The similarity in transcriptional footprints across cancers suggested that tumor subtypes are commonly unified by a limited number of molecular themes.

The landscape and therapeutic relevance of cancer-associated transcript fusions.

Transcript fusions as a result of chromosomal rearrangements have been a focus of attention in cancer as they provide attractive therapeutic targets. To identify novel fusion transcripts with the potential to be exploited therapeutically, we analyzed RNA sequencing, DNA copy number and gene mutation data from 4366 primary tumor samples. To avoid false positives, we implemented stringent quality criteria that included filtering of fusions detected in RNAseq data from 364 normal tissue samples. Our analysis identified 7887 high confidence fusion transcripts across 13 tumor types. Our fusion prediction was validated by evidence of a genomic rearrangement for 78 of 79 fusions in 48 glioma samples where whole-genome sequencing data were available. Cancers with higher levels of genomic instability showed a corresponding increase in fusion transcript frequency, whereas tumor samples harboring fusions contained statistically significantly fewer driver gene mutations, suggesting an important role for tumorigenesis. We identified at least one in-frame protein kinase fusion in 324 of 4366 samples (7.4%). Potentially druggable kinase fusions involving ALK, ROS, RET, NTRK and FGFR gene families were detected in bladder carcinoma (3.3%), glioblastoma (4.4%), head and neck cancer (1.0%), low-grade glioma (1.5%), lung adenocarcinoma (1.6%), lung squamous cell carcinoma (2.3%) and thyroid carcinoma (8.7%), suggesting a potential for application of kinase inhibitors across tumor types. In-frame fusion transcripts involving histone methyltransferase or histone demethylase genes were detected in 111 samples (2.5%) and may additionally be considered as therapeutic targets. In summary, we described the landscape of transcript fusions detected across a large number of tumor samples and revealed fusion events with clinical relevance that have not been previously recognized. Our results support the concept of basket clinical trials where patients are matched with experimental therapies based on their genomic profile rather than the tissue where the tumor originated.

The role of chromophore in the lipid-protein interactions in bacteriorhodopsin-phosphatidylcholine vesicles.

By fluorescence and phase properties of a 1-acyl-2-[8-(2-anthroyl)-octanoyl]-sn-glycero-3-phosphocholine probe, the influence of the chromophore on the phase transition of bacteriorhodopsin-lipid vesicles was investigated. It was observed that removal of the chromophore led to the down-shifting of the phase transition temperatures. The temperatures corresponding to the beginning and ending of the gel-liquid phase transition were also influenced. This demonstrated that the liquid phase is reached more easily when the chromophore is bleached. The results indicate that removal of the chromophore alters the protein-lipid interactions. It is suggested that this alteration might be related to the change in the lipid molecular packing.