Endostaples: are they the solution to graft migration and Type I endoleaks?
Effective proximal sealing, especially in the long-term, remains a limitation of contemporary endovascular aortic aneurysm repair (EVAR). Endostaples that fixate the proximal stent-graft to the aortic neck wall, aiming for better apposition and proximal sealing have been recently introduced in clinical practice to address this problem. Initial experimental studies have shown that endostaples can increase proximal stent-graft fixation to levels equivalent or superior to that of a hand-sewn anastomosis. Further clinical studies aimed to investigate whether this increased proximal fixation results in reduced migration and better sealing with lower rates of type I endoleak. The present chapter discusses the efficacy of endostaples in reducing migration and type I endoleak after EVAR, based on published clinical data.


Rapid Room-Temperature Gelation of Crude Oils by a Wetted Powder Gelator.
Phase-selective organogelators (PSOGs) not only exhibit ability to phase-selectively congeal oil from oily water but also allow easy separation of gelled oil from the body of water. However, all hitherto reported PSOGs either necessitate carrier solvents for their dissolution or suffer from an extremely slow action in gelling oil in the powder form. A previously unexplored generally applicable wetting strategy is now described to dramatically enhance, by up to two orders of magnitude, gelling speed of the resultant wet but non-sticky gelator in the powder form in crude oils of widely ranging viscosities. Such unprecedented rapid gelling speeds enable rapid gelation of six types of (un)weathered crude oils within minutes at room temperature, making PSOGs one step closer to their eventual practical uses as one of important oil spill control technologies.


Structural and functional characterisation of ferret interleukin-2.
While the ferret is a valuable animal model for a number of human viral infections, such as influenza, Hendra and Nipah, evaluating the cellular immune response following infection has been hampered by the lack of a number of species-specific immunological reagents. Interleukin 2 (IL-2) is one such key cytokine. Ferret recombinant IL-2 incorporating a C-terminal histidine tag was expressed and purified and the three-dimensional structure solved and refined at 1.89 Å by X-ray crystallography, which represents the highest resolution and first non-human IL-2 structure. While ferret IL-2 displays the classic cytokine fold of the four-helix bundle structure, conformational flexibility was observed at the second helix and its neighbouring region in the bundle, which may result in the disruption of the spatial arrangement of residues involved in receptor binding interactions, implicating subtle differences between ferret and human IL-2 when initiating biological functions. Ferret recombinant IL-2 stimulated the proliferation of ferret lymph node cells and induced the expression of mRNA for IFN-γ and Granzyme A.


Hepatitis C Virus Infection in a Hemodialysis and Kidney Transplant Patient: An Eight Year Follow-up Report.
We report our experience in the management of a hemodialysis (HD) patient who acquired hepatitis C virus (HCV) infection while on dialysis, and subsequently received a kidney transplant. The potential role of alpha-interferon in the management of HCV infection is discussed, as well as the potential for azathioprine to perpetuate HCV induced liver disease following kidney transplantation. The management of this patient summarizes our standard practice for the management of HCV infection during HD and following kidney transplantation.


Cerebellar Changes in Guinea Pig Offspring Following Suppression of Neurosteroid Synthesis During Late Gestation.
Elevated gestational concentrations of allopregnanolone are essential for the development and neuroprotection of the foetal brain. Preterm birth deprives the foetus of these high levels of allopregnanolone, which may contribute to the associated adverse effects on cerebellar development. Preterm birth alters expression of GABAA receptor subunit composition, which may further limit neurosteroid action. The objective of this study was to determine the effects of suppression of allopregnanolone levels on the markers of development and functional outcome. Pregnant guinea pigs were treated with finasteride at a dose (25 mg/kg maternal weight) shown to suppress allopregnanolone between 60 days of gestation until delivery (term ∼71 days). The cerebella from neonates, whose mothers were treated with finasteride or vehicle during pregnancy, were collected at postnatal age 8. Pups that received finasteride displayed significantly greater glial fibrillary acid protein area coverage and reduced GABAA receptor α6-subunit messenger RNA within the cerebellum than pups that were exposed to vehicle. These findings indicate that loss of neurosteroid action on the foetal brain in late gestation produces prolonged astrocyte activation and reductions in GABAA receptor α6-subunit expression. These changes may contribute to the long-term changes in function associated with preterm birth.


[Combined effect of cosolvent and cyclodextrin on solubilization of insoluble drugs].
To investigate the combined effect of cosolvent and cyclodextrin (CD) on solubilization of insoluble drugs. Phase-solubility method was applied to determine solubilization of two diterpenoids in cosolvent / cyclodextrin combinations. The combined effect was evaluated and explained with an established mathematical model, and the model parameters were calculated by means of nonlinear regression analysis. The strong agreement between the predicted and the observed solubility data supports the validity of the proposed model, with the determination coefficients of two regression models were 0.993 and 0.992, separately. The validated mathematical model can be used to explain and predict the combined solubilization of the two insoluble drugs in different cosolvent systems.


Prenatal genetic diagnosis in a triplet gestation.
The increasing incidence of multiple gestations and the need for genetic diagnosis often pose a question as to the safety of amniocentesis in these cases. Under careful sonographic control and using indigo carmine as a marker, a technique for amniocentesis in a triplet gestation is presented.


Probing the intrinsic features and environmental stabilization of multiply charged anions.
Multiply charged anions (MCAs) represent exotic, highly energetic species in the gas-phase due to their propensity to undergo unimolecular decay via electron loss or ionic fragmentation. There is considerable fundamental interest in these systems since they display novel potential energy surfaces that are characterized by Coulomb barriers. Over recent years, considerable progress has been made in understanding the factors that affect the stability, decay pathways and reactivity of gas-phase MCAs, mainly as a result of the application of electrospray ionization as a generic technique for transferring solution-phase MCAs into the gas-phase for detailed characterization. We review contemporary work in this field, focusing on the factors that control the intrinsic stability of MCAs, both as isolated gas-phase ions, and on their complexation with solvent molecules and counter-ions. While studies of MCAs are primarily of fundamental interest, several classes of important biological ions are commonly observed as MCAs in the gas-phase (e.g. oligonucleotides, sugars). Recent results for biologically relevant ions are emphasised, since a fundamental understanding of the properties of gas-phase MCAs will be highly valuable for developing further analytical methods to study these important systems.


Online Acceptance and Commitment Therapy for People with Painful Diabetic Neuropathy in the United Kingdom: A Single-Arm Feasibility Trial.
This study aimed to assess the feasibility of online Acceptance and Commitment Therapy for painful diabetic neuropathy in the United Kingdom and to determine if a larger randomized controlled trial testing treatment efficacy is justified. Participants with painful diabetic neuropathy were recruited online and from hospital services. This was a single-arm study in which all participants received online Acceptance and Commitment Therapy. Participants completed questionnaires at baseline and three months post-treatment. Primary feasibility outcomes were recruitment, retention, and treatment completion rates. Secondary outcomes were pre- to post-treatment effects on pain outcomes and psychological flexibility. Of 225 potentially eligible participants, 30 took part in this study. Regarding primary feasibility outcomes, the treatment completion and follow-up questionnaire completion rates were 40% and 100%, respectively. Generally, at baseline those who completed the treatment, compared with those who did not, had better daily functioning and higher psychological flexibility. With respect to secondary outcomes, results from the completers group showed clinically meaningful effects at post-treatment for 100% of participants for pain intensity and pain distress, 66.7% for depressive symptoms, 58.3% for functional impairment, 41.7% for cognitive fusion, 66.7% for committed action, 58.3% for self-as-context, and 41.7% for pain acceptance. This preliminary trial suggests feasibility of recruitment and follow-up questionnaire completion rates, supporting planning for a larger randomized controlled trial. However, treatment completion rates did not achieve the prespecified feasibility target. Changes to the treatment content and delivery may enhance the feasibility of online Acceptance and Commitment Therapy for people with painful diabetic neuropathy on a larger scale.


Chimeric studies of the extracellular domain of the rat thyrotropin (TSH) receptor: amino acids (268-304) in the TSH receptor are involved in ligand high affinity binding, but not in TSH receptor-specific signal transduction.
A series of chimeric TSH-LH/CG receptors were constructed by substituting homologous segments of the extracellular domain of the rat TSH receptor with corresponding segments of rat LH/CG receptor: C1 (amino acids 37-123 substituted), C2 (91-112), C3 (173-234), C4 (233-266), C5 (268-304), C6 (112-305) and C7 (36-404). After transfection in Cos-7 cell, TSH- and LH/CG-receptor activities of these chimeras were evaluated and compared with those of deletion mutants involving the same residues [Kosugi et al. Thyroid 1:321 (1991)]. Western blot analyses revealed that most of the chimeric receptor proteins were normally synthesized and integrated in the membrane of transfected Cos-7 cells: an antibody to a TSH receptor specific synthetic peptide (residues 352-366) identified 170-190kDa and 90-100kDa TSH receptor structures in the plasma membrane fractions of Cos-7 cells transfected with wild-type TSH receptor cDNA and the C1 to C6 chimeras, but not C7 or wild LH/CG receptor cDNA. Despite this, no receptor except C5 exhibited any significant TSH receptor activities either in [12I]TSH binding or in cAMP responses to TSH and thyroid-stimulating antibodies (TSAbs) from Graves' patients. The chimeric receptor C5 exhibited only low affinity TSH binding (Kd = 3.5 x 10(-8) M), as did its counterpart the M2C mutant with residues 268-304 deleted. However, unlike M2C, C5 demonstrated a significant cAMP response to TSH as well as to TSAbs. The cAMP increase in response to TSH in the wild type receptor was observed at 10(-11) M TSH. In C5 the response was first evident at 10(-10) M TSH, but the maximum cAMP stimulation by TSH and TSAbs in C5 (EC50 = 6.7 x 10(-10) M) was approximately the same as the wild type receptor (EC50 = 1.5 x 10(-10) M). Inhibition of either TSH- or TSAb- stimulated cAMP increase by thyroid-stimulating blocking antibodies (TSBAbs) was also preserved in C5. These results suggest that amino acids 268-304 do not include an important determinant required for signal transduction, since a significant cAMP response to TSH and TSAbs was observed in the C5 receptor with these residues substituted. Additionally, these residues appear to be involved in ligand high affinity binding because high affinity TSH binding was lost in the chimeric receptor C5.


Human herpesvirus-8 encoded Kaposin: subcellular localization using immunofluorescence and biochemical approaches.
Human herpesvirus-8 (HHV-8) has been causally linked to the development of Kaposi's sarcoma (KS). DNA sequence analysis of the viral genome revealed a total of 81 open reading frames (ORF). Interestingly, only a small subset of these ORFs has been shown to be transcribed in cells latently infected with HHV-8 and in cells of the KS lesions. Among the genes active during latency, kaposin, is noted for its abundance and ability to transform cells in culture, thus implicating a potential role in KS pathogenesis. This has prompted us to undertake an investigation on elucidating the mechanism(s) by which Kaposin brings about transformation of cells. Towards this goal, we have generated an eukaryotic expression plasmid encoding Kaposin (Kap). As Kaposin is predicted to be a type II membrane protein, several strategies were utilized to address this, including the generation of Kaposin with the Flag (FL) epitope (DYKDDDDK) at the C-terminus of the protein (Kap-C-FL). Antibodies specific for Kaposin (kap-2), recognized both Kaposin and Kaposin-Flag, while antibodies against the Flag epitope recognized only Kaposin-Flag. Transfection of Kap and Kap-C-FL expression plasmid DNA into NIH3T3 cells resulted in cellular clones that exhibited a phenotypic property of transformation by forming large, multiclustered cells, when grown on soft agar. Because there is controversial data regarding the localization of Kaposin in cells, we examined the subcellular localization of Kaposin using confocal microscopy. We observed that Kaposin and Kaposin-Flag showed an intense staining surrounding the nucleus. Although there was no staining at the cell membrane of transfected cells, FACS analysis using kap-2 or Flag antibodies, under nonpermeable conditions, showed positivity. Cell fractionation studies further showed that the majority of Kaposin was detected in the nuclear fraction by Western blot analysis. The cytoplasmic and detergent soluble membrane fractions did not show Kaposin protein; however, a small amount was detected in the detergent insoluble membrane fraction. Taken together, these results suggest that Kaposin exhibits multicompartmental localization in cells.


Bilateral tonic-clonic seizure and focal cortical hyperexcitability in familial Creutzfeldt-Jakob disease with E200K mutation of the prion protein.
Convulsive epileptic seizures are rare in Creutzfeldt-Jakob disease (CJD), and their clinical and EEG features have not been reported in detail. We describe a case of familial CJD with an E200K mutation of the prion protein who presented with bilateral tonic-clonic seizures (BTCS) during long-term video-EEG monitoring. Semiologically, BTCS showed focal clinical signs such as head turning and eye deviation to the left. The ictal EEG started with generalized polyspikes. Interictal EEG showed generalized periodic discharges with right fronto-temporal predominance (larger amplitude and earlier onset compared with other regions). MRI showed high-intensity signals persistently in the right temporo-parietal region on diffusion-weighted images (DWI). Interictal single-photon emission computed tomography (SPECT) showed hyperperfusion in the same region. Brain pathology revealed typical spongiform changes in CJD without other pathological findings of rapidly progressive dementia. Our case demonstrates that CJD can cause BTCS with generalized EEG changes and focal semiological/imaging abnormalities, suggesting that diffuse and inhomogeneous cortical and subcortical epileptic networks may develop in familial CJD.


Fas death receptor enhances endocytic membrane traffic converging into the Golgi region.
The death receptor Fas/CD95 initiates apoptosis by engaging diverse cellular organelles including endosomes. The link between Fas signaling and membrane traffic has remained unclear, in part because it may differ in diverse cell types. After a systematic investigation of all known pathways of endocytosis, we have clarified that Fas activation opens clathrin-independent portals in mature T cells. These portals drive rapid internalization of surface proteins such as CD59 and depend upon actin-regulating Rho GTPases, especially CDC42. Fas-enhanced membrane traffic invariably produces an accumulation of endocytic membranes around the Golgi apparatus, in which recycling endosomes concentrate. This peri-Golgi polarization has been documented by colocalization analysis of various membrane markers and applies also to active caspases associated with internalized receptor complexes. Hence, T lymphocytes show a diversion in the traffic of endocytic membranes after Fas stimulation that seems to resemble the polarization of membrane traffic after their activation.


Expression of epidermal growth factor and epidermal growth factor receptor genes in healing colonic anastomoses in rats.
This study uses complementary DNA probes to find out if epidermal growth factor (EGF) and EGF receptor genes are activated in healing colonic anastomoses in rats. 35 rats had a colocolonic anastomosis which was removed for study 1, 2, 3, 5, 7, or 14 days after operation. Six animals without surgery served as controls. The specimens were examined by northern blotting, in situ hybridisation, and conventional microscopy. Microscopic healing progressed normally. The expression of the EGF gene was minimal and surgery did not activate it. However, surgical trauma did increase the expression of the EGF receptor gene by 2.2 times when compared to day one. In situ hybridisations localised a strong EGF receptor expression in mucosal epithelial cells in all specimens, and a moderate reaction in fibroblasts in the repair tissue in the anastomotic line. The enhanced EGF receptor gene expression suggests that anastomotic healing is associated with the presence of EGF or an EGF-like substance and its activity increases during the first postoperative week.


Preparation and certification of standard reference material 3278 tocopherols in edible oils.
Standard Reference Material (SRM) 3278 Tocopherols in Edible Oils has been issued for use as a quality assurance tool in the measurement of tocopherols. Like other natural-matrix SRMs, this material can be used in method validation or in assignment of tocopherol values to in-house quality control materials. Because most edible oils contain one predominant tocopherol isoform, the SRM is a blend of sunflower, soy, canola, and safflower oils to provide roughly comparable chromatographic peak heights of the two main tocopherols, γ and α, with smaller amounts of δ and β. The four tocopherol isoforms were determined by three independent liquid chromatography methods with absorbance and fluorescence detection. Various chromatographic and detection modes are used for assignment of certified values because biases inherent to one method should not be present in the other, and the existence of bias can therefore be identified.


Role of murine Peyer's patch lymphocytes against primary and challenge infections with Cryptosporidium parvum.
In order to determine the role of Peyeros patch lymphocytes (PPL) in self-clearing of Cryptosporidium parvum infection in murine models, changes in PPL subsets, their cytokine expression, and in vitro IgG1 and IgA secretions by PPL were observed in primary- and challenge-infected C57BL/6 mice. In primary-infected mice, the percentages of CD4+ T cells, CD8+ T cells, sIgA+ B cells, IL-2+ T cells, and IFN-gamma+ T cells among the PPL, increased significantly (P < 0.05) on day 10 post-infection (PI). Secretion of IgG1 and IgA in vitro by PPL also increased on day 10 PI. However, all these responses, with the exception of IgG1 and IgA secretions, decreased in challenge-infected mice on day 7 post-challenge (= day 13 PI); their IgG1 and IgA levels were higher (P > 0.05) than those in primaryinfected mice. The results suggest that murine PPL play an important role in self-clearing of primary C. parvum infections through proliferation of CD4+, CD8+, IL-2+, and IFN-gamma+ T cells, and IgG1 and IgA-secreting B cells. In challenge infections, the role of T cells is reduced whereas that of B cells secreting IgA appeared to be continuously important.


Influence of surface cracks on laser-induced damage resistance of brittle KH₂PO₄ crystal.
Single point diamond turning (SPDT) currently is the leading finishing method for achieving ultra-smooth surface on brittle KH(2)PO(4) crystal. In this work, the light intensification modulated by surface cracks introduced by SPDT cutting is numerically simulated using finite-difference time-domain algorithm. The results indicate that the light intensification caused by surface cracks is wavelength, crack geometry and position dependent. Under the irradiation of 355 nm laser, lateral cracks on front surfaces and conical cracks on both front and rear surfaces can produce light intensification as high as hundreds of times, which is sufficient to trigger avalanche ionization and finally lower the laser damage resistance of crystal components. Furthermore, we experimentally tested the laser-induced damage thresholds (LIDTs) on both crack-free and flawed crystal surfaces. The results imply that brittle fracture with a series of surface cracks is the dominant source of laser damage initiation in crystal components. Due to the negative effect of surface cracks, the LIDT on KDP crystal surface could be sharply reduced from 7.85J/cm(2) to 2.33J/cm(2) (355 nm, 6.4 ns). In addition, the experiment of laser-induced damage growth is performed and the damage growth behavior agrees well with the simulation results of light intensification caused by surface cracks with increasing crack depths.


Minimally Invasive Robotic Laser Corpus Callosotomy: A Proof of Concept.
We describe the feasibility of using minimally invasive robotic laser interstitial thermotherapy (LITT) for achieving an anterior two-thirds as well as a complete corpus callosotomy. Ten probe trajectories were plotted on normal magentic resonance imaging (MRI) scans using the Brainlab Stereotactic Planning Software (Brainlab, Munich, Germany). The NeuroBlate® System (Monteris Medical, MN, USA) was used to conform the thermal burn to the corpus callosum along the trajectory of the probe. The distance of the ideal entry site from either the coronal suture and the torcula or nasion and the midline was calculated. The distance of the probe tip from the dorsal and ventral limits of the callosotomy in the sagittal plane were also calculated. Anterior two-thirds callosotomy was possible in all patients using a posterior parieto-occipital paramedian trajectory through the non-dominant lobe. The average entry point was 3.64 cm from the midline, 10.6 cm behind the coronal suture, and 9.2 cm above the torcula. The probe tip was an average of 1.4 cm from the anterior commissure. For a total callosotomy, an additional contralaterally placed frontal probe was used to target the posterior one-third of the corpus callosum. The average entry site was 3.3 cm from the midline and 9.1 cm above the nasion. The average distance of the probe tip from the base of the splenium was 0.94 cm. The directional thermoablation capability of the NeuroBlate® system allows for targeted lesioning of the corpus callosum, to achieve a two-thirds or complete corpus callosotomy. A laser distance of < 2 cm is sufficient to reach the entire corpus callosum through one trajectory for an anterior two-thirds callosotomy and two trajectories for a complete callosotomy.


Turtles as a Possible Reservoir of Nontyphoidal Salmonella in Shanghai, China.
Terrapins and turtles are known to transmit Salmonella to humans. However, little was known about the occurrence of this pathogen in soft-shelled terrapin that is a popular delicacy in Chinese and other East Asian cuisines. We isolated and characterized 82 (24.4%) isolates of Salmonella from 336 fecal samples of soft-shelled terrapins (51 of 172; 29.7%) and pet turtles (31 of 164; 18.9%) in Shanghai. Salmonella Thompson was the most common serotype (17.1%) among others. Many isolates (84.1%) were resistant to multiple antimicrobials (≥3). Molecular analysis of Salmonella Thompson and Salmonella Typhimurium using pulsed-field gel electrophoresis unveiled a close genetic relationship between several human and terrapin isolates. Our results highlight the risk associated with the handling and consumption of turtles and their role in the spread of Salmonella in the human salmonellosis.


Pulmonary function of exposed and control workers in a Nigerian nonsoapy detergent factory.
Respiratory symptoms and spirometric pulmonary function data [i.e., first-second forced expiratory volume (FEV1.0) and forced vital capacity (FVC)] for 128 (30%) males who were exposed to alkyl benzene sulphonate in a detergent factory and for 56 (76%) unexposed workers in the same factory are reported herein. Exposed subjects had been employed for 1 month to 15 yr, and they generally complained of cough and mucus secretions, nasal catarrh, chest pain, and breathlessness. Unexposed workers had been employed for 1 month to 13 yr and had a significantly lower (P less than .001) frequency of symptoms, as well as significantly higher (.01 greater than P greater than .001) FEV1.0 and FVC than the exposed workers. The reduction in pulmonary function of exposed subjects from the predicted was significantly higher (.01 greater than P greater than .001) than that experienced by the unexposed subjects. There was a significant 8-hr workshift depression in lung function. There was radiological evidence of pulmonary fibrosis, but lack of pre-employment chest radiographs renders this inconclusive. Respiratory symptoms in exposed subjects decreased with duration of employment, which probably indicates the exodus from the work force of those who could not tolerate the nonsoapy detergent.


[Optimization of PET image quality by means of 3D data acquisition and iterative image reconstruction].
In the recent past, several algorithms have been developed in order to transform 3D sinograms acquired at volume PET systems into 2D data sets. These methods offer the possibility to combine the high sensitivity of the 3D measurement with the advantages of iterative 2D image reconstruction. The purpose of our study was the assessment of this approach by using phantom measurements and patient examinations. The experiments were performed at the latest-generation whole-body PET system ECAT EXACT HR+. For 2D data acquisition, a collimator of thin tungsten septa was positioned in the field-of-view. Prior to image reconstruction, the measured 3D data were sorted into 2D sinograms by suing the Fourier rebinning (FORE) algorithm developed by M. Defrise. The standard filtered backprojection (FBP) method and an optimized ML/EM algorithm with overrelaxation for accelerated convergence were employed for image reconstruction. The spatial resolution of both methods as well as the convergence and noise properties of the ML/EM algorithm were studied in phantom measurements. Furthermore, patient data were acquired in the 2D mode as well as in the 3D mode and reconstructed with both techniques. At the same spatial resolution, the ML/EM-reconstructed images showed fewer and less prominent artefacts than the FBP-reconstructed images. The resulting improved detail conspicuously was achieved for the data acquired in the 2D mode as well as in the 3D mode. The best image quality was obtained by iterative 2D reconstruction of 3D data sets which were previously rebinned into 2D sinograms with help of the FORE algorithm. The phantom measurements revealed that 50 iteration steps with the optimized ML/EM algorithm were sufficient to keep the relative quantitation error below 5%. Our measurements show that the image quality in 3D PET can be improved by using iterative reconstruction techniques. The concept of 3D data acquisition and combining the FORE algorithm with 2D ML/EM reconstruction can readily be employed in clinical practice since the computation time is not considerably longer than that in iterative reconstruction of true 2D data.


Insights into the historical assembly of East Asian subtropical evergreen broadleaved forests revealed by the temporal history of the tea family.
Subtropical evergreen broadleaved forests (EBLFs) inhabit large areas of East Asia. Although paleovegetation reconstructions have revealed that the subtropical EBLFs existed in Southwest China during the Miocene, the historical construction of these forests remains poorly known. Here, we used the tea family (Theaceae), a characteristic component of the subtropical EBLFs, to gain new insights into the assembly of this important biome. Using a robust phylogenetic framework of Theaceae based on plastome and nuclear ribosomal DNA sequence data, the temporal history of the family was reconstructed. Data from other characteristic components of subtropical EBLFs, including Fagaceae, Lauraceae and Magnoliaceae, were also integrated. Most of the essential elements of the subtropical EBLFs appear to have originated around the Oligocene-Miocene (O-M) boundary. However, small woody lineages (e.g. Camellia, Hartia) from Theaceae were dated to the late Miocene. Accelerated net diversification rates within Theaceae were also detected near the O-M transition period and the late Miocene. Our results suggest that two independent intensifications of the East Asian summer monsoon (EASM) around the O-M boundary and the late Miocene may have facilitated the historical assembly of the subtropical EBLFs in East Asia.


Bromodomain-Containing Protein 4 (BRD4) Inhibition Sensitizes Palomid 529-Induced Anti-Renal Cell Carcinoma Cell Activity in Vitro and in Vivo.
Mammalian target of rapamycin (mTOR) is a valuable treatment target of renal cell carcinoma (RCC). Palomid 529 is a novel mTORC1/2 dual inhibitor. RCC cells were treated with different concentrations of Palomid 529. Cell survival was tested by MTT assay and clonogenicity assay. Cell proliferation was tested by BrdU ELISA assay. Cell apoptosis was tested by the Hoechst-33342 nuclei staining assay and Histone DNA ELISA assay. mTOR signaling was tested by Western blotting assay and co-immunoprecipitation (IP) assay. The SCID mouse 786-O xenograft model was established to test RCC cell growth in vivo. Palomid 529 exerted cytotoxic, anti-proliferative and pro-apoptotic activities in 786-O RCC cells. Palomid 529 disassembled mTORC1/2, causing de-phosphorylation of mTORC1/2 substrates. Bromodomain-containing protein 4 (BRD4) is a primary resistant factor of Palomid 529. Palomid 529-induced 786-O cell apoptosis was sensitized by BRD4 inhibitors or BRD4 silencing, but inhibited with BRD4 over-expression. Palomid 529-induced cytotoxicity in the primary human RCC cells was negatively correlated with BRD4 expression level. In vivo, Palomid 529 i.p. administration inhibited 786-O xenograft tumor growth in SCID mice. Its anti-tumor activity was further sensitized by co-administration of the BRD4 inhibitor JQ1. Cconclusion: Palomid 529 inhibits RCC cell growth in vitro and in vivo. BRD4 inhibition could further sensitize Palomid 529 against RCC cells.


Time-dependent close-coupling calculations of dielectronic capture in He.
We report on the first time-dependent close-coupling calculation of dielectronic capture into a doubly excited state of a two-electron atom. An incoming electron is represented by a Gaussian wave packet which collides with singly ionized helium in its ground state. The close-coupling equations describe the propagation of the total compound wave function on a two-dimensional radial lattice. By projecting this wave function onto a doubly excited state of neutral helium, we can determine the probability amplitude for dielectronic capture into one of these states and the subsequent autoionization from it.


A comparative clinical, pathological, biochemical and genetic study of fused in sarcoma proteinopathies.
Neuronal intermediate filament inclusion disease and atypical frontotemporal lobar degeneration are rare diseases characterized by ubiquitin-positive inclusions lacking transactive response DNA-binding protein-43 and tau. Recently, mutations in the fused in sarcoma gene have been shown to cause familial amyotrophic lateral sclerosis and fused in sarcoma-positive neuronal inclusions have subsequently been demonstrated in neuronal intermediate filament inclusion disease and atypical frontotemporal lobar degeneration with ubiquitinated inclusions. Here we provide clinical, imaging, morphological findings, as well as genetic and biochemical data in 14 fused in sarcoma proteinopathy cases. In this cohort, the age of onset was variable but included cases of young-onset disease. Patients with atypical frontotemporal lobar degeneration with ubiquitinated inclusions all presented with behavioural variant frontotemporal dementia, while the clinical presentation in neuronal intermediate filament inclusion disease was more heterogeneous, including cases with motor neuron disease and extrapyramidal syndromes. Neuroimaging revealed atrophy of the frontal and anterior temporal lobes as well as the caudate in the cases with atypical frontotemporal lobar degeneration with ubiquitinated inclusions, but was more heterogeneous in the cases with neuronal intermediate filament inclusion disease, often being normal to visual inspection early on in the disease. The distribution and severity of fused in sarcoma-positive neuronal cytoplasmic inclusions, neuronal intranuclear inclusions and neurites were recorded and fused in sarcoma was biochemically analysed in both subgroups. Fused in sarcoma-positive neuronal cytoplasmic and intranuclear inclusions were found in the hippocampal granule cell layer in variable numbers. Cortical fused in sarcoma-positive neuronal cytoplasmic inclusions were often 'Pick body-like' in neuronal intermediate filament inclusion disease, and annular and crescent-shaped inclusions were seen in both conditions. Motor neurons contained variable numbers of compact, granular or skein-like cytoplasmic inclusions in all fused in sarcoma-positive cases in which brainstem and spinal cord motor neurons were available for study (five and four cases, respectively). No fused in sarcoma mutations were found in any cases. Biochemically, two major fused in sarcoma species were found and shown to be more insoluble in the atypical frontotemporal lobar degeneration with ubiquitinated inclusions subgroup compared with neuronal intermediate filament inclusion disease. There is considerable overlap and also significant differences in fused in sarcoma-positive pathology between the two subgroups, suggesting they may represent a spectrum of the same disease. The co-existence of fused in sarcoma-positive inclusions in both motor neurons and extramotor cerebral structures is a characteristic finding in sporadic fused in sarcoma proteinopathies, indicating a multisystem disorder.


Multiple idiopathic external root resorption. A case report.
An unusual case of multiple idiopathic external apical root resorption affecting all four quadrants, in a patient with a history of narcotic intravenous drug addiction and liver disease, is reported. The literature relating to a systemic etiology for external root resorption is briefly reviewed.


Computational analysis of nascent peptides that induce ribosome stalling and their proteomic distribution in Saccharomyces cerevisiae.
Interactions between the ribosomal exit tunnel and the nascent peptide can affect translation elongation rates. While previous studies have already demonstrated the feasibility of such interactions, little is known about the nature of the stalling peptide sequences and their distribution in the proteome. Here we ask which peptide sequences tend to occupy the tunnel of stalled ribosomes and how they are distributed in the proteome. Using computational analysis of ribosome profiling data from S. cerevisiae, we identified for the first time dozens of short stalling peptide sequences and studied their statistical properties. We found that short peptide sequences associated with ribosome stalling tend significantly to be either over- or underrepresented in the proteome. We then showed that the stalling interactions may occur at different positions along the length of the tunnel, prominently close to the P-site. Our findings throw light on the determinants of nascent peptide-mediated ribosome stalling during translation elongation and support the novel conjecture that mRNA translation affects the proteomic distribution of short peptide sequences.


High-level expression of human superoxide dismutase in the cyanobacterium Anacystis nidulans 6301.
A chemically synthesized gene encoding human CuZn superoxide dismutase (hSOD) was cloned into the shuttle vector pBAX18R and expressed in Anacystis nidulans 6301 (Synechococcus sp. strain PCC 6301) under the control of a ribulose-1,5-bisphosphate carboxylase/oxygenase gene (rbc) promoter derived from A. nidulans 6301. The sequences immediately upstream from the hSOD coding region and the distances between the ribosomal binding site and ATG initiation codon strongly affected the expression of the hSOD gene in A. nidulans cells. Optimal expression of hSOD was obtained with the expression vector pBAXSOD8-I, which contained a GGAGAG sequence. In defined conditions, irradiation with light increased hSOD enzyme activity in the transformants > 18-fold and the level of the hSOD protein reached a value of about 3% of the total soluble protein. The transformants that expressed hSOD acquired the ability to extenuate photooxidative damage induced by methyl viologen.


Inducible and myocyte-specific inhibition of PKCalpha enhances cardiac contractility and protects against infarction-induced heart failure.
Mice null for the gene encoding protein kinase Calpha (Prkca), or mice treated with pharmacologic inhibitors of the PKCalpha/beta/gamma isoforms, show an augmentation in cardiac contractility that appears to be cardioprotective. However, it remains uncertain if PKCalpha itself functions in a myocyte autonomous manner to affect cardioprotection in vivo. Here we generated cardiac myocyte-specific transgenic mice using a tetracycline-inducible system to permit controlled expression of dominant negative PKCalpha in the heart. Consistent with the proposed function of PKCalpha, induction of dominant negative PKCalpha expression in the adult heart enhanced baseline cardiac contractility. This increase in cardiac contractility was associated with a partial protection from long-term decompensation and secondary dilated cardiomyopathy after myocardial infarction injury. Similarly, Prkca null mice were also partially protected from infarction-induced heart failure, although the area of infarction injury was identical to controls. Thus, myocyte autonomous inhibition of PKCalpha protects the adult heart from decompensation and dilated cardiomyopathy after infarction injury in association with a primary enhancement in contractility.


Dentin bond strength of Dyract Cem.
To evaluate the influence of provisional cements and of a dentin bonding agent on the adhesion of a self-curing polyacid-modified resin composite (Dyract Cem) on pressurized human dentin. Freshly prepared dentin specimens of human teeth were perfused with physiologic saline. Three different types of surface treatment were evaluated. (1) Dyract Cem was applied to freshly prepared dentin without (Group 1.1) or with (Group 1.2) the respective dentin bonding agent (Prime & Bond 2.0) and cured. (2) Freegenol (Groups 2.1 and 2.2) or Temp Bond (Groups 2.3 and 2.4) were applied first on the dentin surface for 24 hours. Only then was Dyract Cem (with or without Prime & Bond 2.0) added onto the dentin after cleaning the dentin surface with pumice. 3) Prime & Bond 2.0 was applied first on the dentin surface and cured. Then Freegenol (Group 3.1) or Temp Bond (Group 3.2) were added for 24 hours. After cleaning with pumice, Prime & Bond 2.0 was applied for a second time on the dentin (= dual application) and finally Dyract Cem was added. As control, a conventional glass ionomer cement (Ketac-Cem Maxicap; Groups 4.1-4.3) was used in a similar way. After 1,500 thermal cycles with constant imitation of intrapulpal pressure, shear bond strengths were measured. Resulting shear bond strength values were displayed by means of a box plot and they were analyzed statistically by Mann-Whitney, Kruskal-Wallis or one way ANOVA tests. Lowest and highest mean shear bond strength values were 0.27 +/- 0.42 MPa (Group 2.4; single use of Prime & Bond 2.0 with prior application of TempBond) and 5.84 +/- 3.36 MPa (Group 3.1; dual use of Prime & Bond 2.0 with intermediate application of Freegenol). A clearly significant difference between groups could only be found when a single or dual use of the dentin bonding agent Prime & Bond 2.0 were combined with either Freegenol (Groups 2.1, 2.2 and 3.1; Kruskal-Wallis: P < 0.01) or Temp Bond (Groups 2.3, 2.4 and 3.2; Kruskal-Wallis: P < 0.001) as additional dentin surface treatment.


Ala-7, His-10 and Arg-12 are crucial amino acids for activity of a synthetically engineered μ-conotoxin.
Cone snail toxins or conotoxins are often small cysteine-rich peptides which have shown to be highly selective ligands for a wide range of ion channels such as voltage-gated sodium channels (Na(V)s). Na(V)s participate in a wide range of electrophysiological processes. Consequently, their malfunction has been associated with numerous diseases. The development of subtype-selective modulators of Na(V)s remains highly important in the treatment of such disorders. In order to expand our knowledge in the search for novel therapeutics to treat Na(V)-related diseases, we explored the field of peptide engineering. In the current study, the impact of well considered point mutations into a bioactive peptide that was found to be a very potent and selective inhibitor of Na(V)s (i.e. Midi R2) was examined. We designed two peptides, named Midi R2[A7G] and Midi R2[H10A, R12A] which have mutations at position 7, and both 10 and 12, respectively. Electrophysiological recordings indicated that an Ala to Gly mutation at position 7 increased IC50-values from the nanomolar range to the micromolar range. For Midi R2[H10A, R12A] at a concentration of 10 μM, activity is even reduced to 0-10% for all of the tested Na(V)-channels. Circular dichroism measurements proved that overall structural conformations did not change. These findings suggest that the minimal space between the second and the third intercysteine loop of Midi R2 is the sequence RRWARDHSR and that His at position 10 and Arg at position 12 are crucial amino acids for the potency and specificity of Midi R2. In this way, new insights into the structure-activity relationships of μ-conotoxins were found.


A comparison of the distributions of the actinides uranium and thorium with the lanthanide gadolinium in the tissues and eggs of Japanese quail: concentrations of uranium in feeds and foods.
Japanese quail were given UCl3 , UO2 (NO3)2, Th(NO3)4, or GdCl3 ( 153Gd -labeled) intravenously in aqueous solution. Distribution of Th among the tissues was as for Gd; distributions of U(III) and U(VI) were markedly different. For example, 18 hr after a 1.5 mumol/100 g dose, accumulations in females were: growing oocytes, U(III) 2.0%, U(VI) 2.4%, Th 27.7%, Gd 44.7%; leg bones, U(III) 12.5%, U(VI) 14.1%, Th 1.2%, Gd 1.4%; liver, U(III) 1.1%, U(VI) 1.1%, Th 44.0%, Gd 40.2%. Whole body losses by 18 hr were: females, U 24%, Th 14%, Gd 4%; males, U 72%, Th 23%, Gd 1%. Cumulative depositions in yolks of eggs laid over 8 days were: U(III) 1.9%, U(VI) 1.7%, Th 57.3%, Gd 46.8%. The distribution of U in quail may be atypical of actinides . Concentrations of U in various feeds, foods, and mineral supplements ranged from 169 micrograms/g in a phosphate fertilizer for farm use to below the lower detectable limit of .01 microgram/g in many foods intended for human use. Two batches of the game bird laying ration supplied to the quail colony contained 3.05 and 4.42 micrograms U/g. Body burdens of 3.5 micrograms U/bird for noninjected quail were attributed to the U content of this feed.


Deoxynivalenol as a new factor in the persistence of intestinal inflammatory diseases: an emerging hypothesis through possible modulation of Th17-mediated response.
Deoxynivalenol (DON) is a mycotoxin produced by Fusarium species which is commonly found in temperate regions worldwide as a natural contaminant of cereals. It is of great concern not only in terms of economic losses but also in terms of animal and public health. The digestive tract is the first and main target of this food contaminant and it represents a major site of immune tolerance. A finely tuned cross-talk between the innate and the adaptive immune systems ensures the homeostatic equilibrium between the mucosal immune system and commensal microorganisms. The aim of this study was to analyze the impact of DON on the intestinal immune response. Non-transformed intestinal porcine epithelial cells IPEC-1 and porcine jejunal explants were used to investigate the effect of DON on the intestinal immune response and the modulation of naive T cells differentiation. Transcriptomic proteomic and flow cytometry analysis were performed. DON induced a pro-inflammatory response with a significant increase of expression of mRNA encoding for IL-8, IL-1α and IL-1β, TNF-α in all used models. Additionally, DON significantly induced the expression of genes involved in the differentiation of Th17 cells (STAT3, IL-17A, IL-6, IL-1β) at the expenses of the pathway of regulatory T cells (Treg) (FoxP3, RALDH1). DON also induced genes related to the pathogenic Th17 cells subset such as IL-23A, IL-22 and IL-21 and not genes related to the regulatory Th17 cells (rTh17) such as TGF-β and IL-10. DON triggered multiple immune modulatory effects which could be associated with an increased susceptibility to intestinal inflammatory diseases.


Infant feeding practices among Sudanese women now living in regional south east Queensland, Australia.
The purpose of this research was to highlight and compare immigrant Sudanese women's infant feeding choices and patterns before and after moving to a regional city in Queensland, Australia. Semi-structured interviews were conducted with 10 Sudanese mothers who had birthed and breastfed babies both in Africa and Toowoomba. This qualitative research project supported previous research indicating a trend for immigrant women's breastfeeding duration to decline when they moved to another country. The outcomes of this research suggest that the reasons for this decline are complex. The authors conclude that a lack of social support, language difficulties and wanting to fit in with particular Western practices are contributing factors.


Hyperthermia and radiation therapy in the treatment of recurrent Merkel cell tumors.
A high incidence of local recurrence, spread to regional lymph nodes, and distant metastases has been reported after surgical excision of Merkel cell tumors (MCT). The use of postoperative radiation therapy and/or chemotherapy is reviewed from the literature. Despite adjuvant treatment, local tumor recurrences frequently develop. Two patients are presented with metastatic MCT recurrent in previously irradiated sites who had excellent clinical responses and local control following retreatment with local hyperthermia in conjunction with low to moderate dose radiation therapy. These patients represent the first reported use of hyperthermia in the management of MCT. The encouraging local responses described suggest a potential role for the use of hyperthermia and concomitant radiation therapy in the treatment of recurrent MCT.


[Cyclosporin A in aplastic anemia in dogs: first results].
Aplastic anemia in the dog is defined as a reduction of marrow activity comprising all three marrow cell lines or selectively of the erythropoiesis. In comparable cases in human medicine an immunogenic etiology is presumed, and a treatment with the immunosuppressant Cyclosporin A is initiated. There are no experiences with this kind of therapy in aplastic anemia of the dog. Four dogs, three with a hypoplasia of the erythropoietic cell line and one with panmyelophthisis, were treated orally for a period of several months with Cyclosporin A 5-10 mg/kg bodyweight bid. In all patients the plasma concentration of Cyclosporin was controlled in regular intervals. Depending on the plasma concentrations the dose was adapted in several cases. Cyclosporin levels between 200 and 600 ng/ml were considered desirable and their efficacy in the therapy of aplastic anemia was examined. In the patient with panmyelophthisis, therapy was not successful. In this case, a marrow intoxication was presumably responsible for the aplastic anemia and the dog was euthanized. In two of the remaining three cases, therapy with Cyclosporin A was successful; the last dog is still being treated with Cyclosporin A at this time.


Chloroplast DNA replication is regulated by the redox state independently of chloroplast division in Chlamydomonas reinhardtii.
Chloroplasts arose from a cyanobacterial endosymbiont and multiply by division. In algal cells, chloroplast division is regulated by the cell cycle so as to occur only once, in the S phase. Chloroplasts possess multiple copies of their own genome that must be replicated during chloroplast proliferation. In order to examine how chloroplast DNA replication is regulated in the green alga Chlamydomonas reinhardtii, we first asked whether it is regulated by the cell cycle, as is the case for chloroplast division. Chloroplast DNA is replicated in the light and not the dark phase, independent of the cell cycle or the timing of chloroplast division in photoautotrophic culture. Inhibition of photosynthetic electron transfer blocked chloroplast DNA replication. However, chloroplast DNA was replicated when the cells were grown heterotrophically in the dark, raising the possibility that chloroplast DNA replication is coupled with the reducing power supplied by photosynthesis or the uptake of acetate. When dimethylthiourea, a reactive oxygen species scavenger, was added to the photoautotrophic culture, chloroplast DNA was replicated even in the dark. In contrast, when methylviologen, a reactive oxygen species inducer, was added, chloroplast DNA was not replicated in the light. Moreover, the chloroplast DNA replication activity in both the isolated chloroplasts and nucleoids was increased by dithiothreitol, while it was repressed by diamide, a specific thiol-oxidizing reagent. These results suggest that chloroplast DNA replication is regulated by the redox state that is sensed by the nucleoids and that the disulfide bonds in nucleoid-associated proteins are involved in this regulatory activity.


A study of transfer and prevalence of organic gunshot residues.
The main goal of the present study was to determine the amounts and distribution of organic gunshot residues (OGSR) on the shooter's upper body and clothing after discharging a pistol. A preliminary study was also performed to evaluate the prevalence of OGSR in the general population as well as in a police laboratory environment. In the transfer study, results indicated that OGSR are not only transferred to the hand of the shooter, but also to other parts of the upper body. Thus, wrists and forearms also represent interesting targets as they are washed less frequently than hands. Samples from the face and hair of the shooters resulted in no OGSR detection just after firing. It was also observed that the concentrations recovered from clothing are generally higher compared to the same skin area. Prevalence in both general (n=27) and police populations (n=25) was low. No OGSR was detected in the samples from the general population and only two samples from the police population were found positive.


A Novel Therapeutic Approach to Treat Alzheimer's Disease by Neurotrophic Support During the Period of Synaptic Compensation.
Alzheimer's disease (AD), at present, is considered an incurable disease and a major dilemma with no drug to stop or slow down its progression. Drugs that are currently available in the market are able to only transiently improve the clinical symptoms. The repeated failures in developing an effective drug has led to the suggestion that the medical intervention was probably too late to be effective since the pathology starts many years before the appearance of the clinical symptoms. Probably, at the time of the appearance of clinical symptoms the brain has undergone major neuronal and synaptic loss. Because of the uncertainty on when to use a prevention therapy, especially targeting amyloid-β (Aβ) and tau pathologies, interventions that rely on the regenerative capacity of the brain such as the modulation of the inherent neurogenesis and neuronal plasticity represent a promising therapeutic strategy. Such an approach can act both at early as well as late stages of the disease and remove the barrier of the time of intervention. In this article, we review studies mainly from our laboratory that show the merit of early intervention during the synaptic and neuronal compensation period where the brain still has the capacity to self-repair by offering neurotrophic support in reversing cognitive impairment, neuronal and synaptic deficits, Aβ, and tau pathologies and decreasing mortality in a transgenic mouse model of AD.


Effects of conformism on the cultural evolution of social behaviour.
Models of cultural evolution study how the distribution of cultural traits changes over time. The dynamics of cultural evolution strongly depends on the way these traits are transmitted between individuals by social learning. Two prominent forms of social learning are payoff-based learning (imitating others that have higher payoffs) and conformist learning (imitating locally common behaviours). How payoff-based and conformist learning affect the cultural evolution of cooperation is currently a matter of lively debate, but few studies systematically analyse the interplay of these forms of social learning. Here we perform such a study by investigating how the interaction of payoff-based and conformist learning affects the outcome of cultural evolution in three social contexts. First, we develop a simple argument that provides insights into how the outcome of cultural evolution will change when more and more conformist learning is added to payoff-based learning. In a social dilemma (e.g. a Prisoner's Dilemma), conformism can turn cooperation into a stable equilibrium; in an evasion game (e.g. a Hawk-Dove game or a Snowdrift game) conformism tends to destabilize the polymorphic equilibrium; and in a coordination game (e.g. a Stag Hunt game), conformism changes the basin of attraction of the two equilibria. Second, we analyse a stochastic event-based model, revealing that conformism increases the speed of cultural evolution towards pure equilibria. Individual-based simulations as well as the analysis of the diffusion approximation of the stochastic model by and large confirm our findings. Third, we investigate the effect of an increasing degree of conformism on cultural group selection in a group-structured population. We conclude that, in contrast to statements in the literature, conformism hinders rather than promotes the evolution of cooperation.


Ex vivo and in vivo studies of CME-1, a novel polysaccharide purified from the mycelia of Cordyceps sinensis that inhibits human platelet activation by activating adenylate cyclase/cyclic AMP.
CME-1, a novel water-soluble polysaccharide, was purified from the mycelia of Cordyceps sinensis, and its chemical structure was characterized to contain mannose and galactose in a ratio of 4:6 (27.6 kDa). CME-1 was originally observed to exert a potent inhibitory effect on tumor migration and a cytoprotective effect against oxidative stress. Activation of platelets caused by arterial thrombosis is relevant to various cardiovascular diseases (CVDs). However, no data are available concerning the effects of CME-1 on platelet activation. Hence, the purpose of this study was to examine the ex vivo and in vivo antithrombotic effects of CME-1 and its possible mechanisms in platelet activation. The aggregometry, immunoblotting, flow cytometric analysis and platelet functional analysis were used in this study. CME-1 (2.3-7.6 μM) exhibited highly potent activity in inhibiting human platelet aggregation when stimulated by collagen, thrombin, and arachidonic acid but not by U46619. CME-1 inhibited platelet activation accompanied by inhibiting Akt, mitogen-activated protein kinases (MAPKs), thromboxane B2 (TxB2) and hydroxyl radical (OH(●)) formation. However, CME-1 interrupted neither FITC-triflavin nor FITC-collagen binding to platelets. CME-1 markedly increased cyclic AMP levels, but not cyclic GMP levels, and stimulated vasodilator-stimulated phosphoprotein (VASP) phosphorylation. SQ22536, an inhibitor of adenylate cyclase, but not ODQ, an inhibitor of guanylate cyclase, obviously reversed the CME-1-mediated effects on platelet aggregation and vasodilator-stimulated phosphoprotein (VASP), Akt, p38 MAPK phosphorylation, and TxB2 formation. CME-1 substantially prolonged the closure time of whole blood and the occlusion time of platelet plug formation. This study demonstrates for the first time that CME-1 exhibits highly potent antiplatelet activity that may initially activate adenylate cyclase/cyclic AMP and, subsequently, inhibit intracellular signals (such as Akt and MAPKs), ultimately inhibiting platelet activation. This novel role of CME-1 indicates that CME-1 exhibits high potential for application in treating and preventing CVDs.


Olfactory Ensheathing Cell Transplantation after a Complete Spinal Cord Transection Mediates Neuroprotective and Immunomodulatory Mechanisms to Facilitate Regeneration.
Multiple neural and peripheral cell types rapidly respond to tissue damage after spinal cord injury to form a structurally and chemically inhibitory scar that limits axon regeneration. Astrocytes form an astroglial scar and produce chondroitin sulfate proteoglycans (CSPGs), activate microglia, and recruit blood-derived immune cells to the lesion for debris removal. One beneficial therapy, olfactory ensheathing cell (OEC) transplantation, results in functional improvements and promotes axon regeneration after spinal cord injury. The lack of an OEC-specific marker, however, has limited the investigation of mechanisms underlying their proregenerative effects. We compared the effects of enhanced green fluorescent protein-labeled fibroblast (FB) and OEC transplants acutely after a complete low-thoracic spinal cord transection in adult rats. We assessed the preservation of neurons and serotonergic axons, the levels of inhibitory CSPGs and myelin debris, and the extent of immune cell activation between 1 and 8 weeks postinjury. Our findings indicate that OECs survive longer than FBs post-transplantation, preserve axons and neurons, and reduce inhibitory molecules in the lesion core. Additionally, we show that OECs limit immune-cell activation and infiltration, whereas FBs alter astroglial scar formation and increase immune-cell infiltration and concomitant secondary tissue damage. Administration of cyclosporine-A to enhance graft survival demonstrated that immune suppression can augment OEC contact-mediated protection of axons and neurons during the first 2 weeks postinjury. Collectively, these data suggest that OECs have neuroprotective and immunomodulatory mechanisms that create a supportive environment for neuronal survival and axon regeneration after spinal cord injury. Spinal cord injury creates physical and chemical barriers to axon regeneration. We used a complete spinal cord transection model and olfactory ensheathing cell (OEC) or fibroblast (FB; control) transplantation as a repair strategy. OECs, but not FBs, intermingled with astrocytes, facilitated astroglial scar border formation and sequestered invading peripheral cells. OECs attenuated immune cell infiltration, reduced secondary tissue damage, protected neurons and axons in the lesion core, and helped clear myelin debris. Immunosuppression enhanced survival of OECs and FBs, but only OEC transplantation promoted scaffold formation in the lesion site that facilitated axon regeneration and neuron preservation.


Effects of protein and fat concentration in coproduct-based growing calf diets on performance and carcass composition.
Angus×Simmental crossbred heifers (n=150) and steers (n=100) were used to evaluate 1 of 5 growing diets: 1) a corn-based growing diet (CRN); 2) a high-fat, high-protein coproduct blend; 3) a high-fat, low-protein coproduct blend; 4) a low-fat, high-protein coproduct blend; and 5) a low-fat, low-protein coproduct blend in a 2×2+1 factorial arrangement. Low-protein and low-fat diets were formulated to be isonitrogenous and isofat to CRN (16.0% CP and 3.0% fat), and high-protein and high-fat diets were formulated to have 20.0% CP and 5.0% fat, respectively. Calves were weaned at 85±1.2 d, blocked by weight, and allotted to pens (10 calves/pen) within sex (10 pens of steers and 15 pens of heifers). The objective of this experiment was to determine if the concentration of protein or fat or their interaction in coproducts used in growing diets fed to early-weaned calves affects feedlot performance or carcass composition. Starting on d 0, calves (141±1.2 d of age) were fed experimental diets for 112 d and then fed a common feedlot diet for an additional 112 d. Body weight, hip height, and ultrasound data were collected at the end of each 112-d feeding phase. Carcass data included HCW, LM area (LMA), 12th-rib back fat (BF), marbling score (MS), KPH, and USDA quality grade. There was no fat×protein interaction (P≥0.27); therefore, only main effects are discussed. No effects (P≥0.47) of CRN, protein, or fat were detected for BW at d 112 or 224. Increased dietary protein resulted in greater (P=0.04) ADG at d 112 compared to calves fed low protein. Feeding cattle CRN decreased (P=0.04) DMI and increased (P<0.01) G:F during the growing phase compared to coproducts. High-fat diets increased (P=0.05) BF in calves at d 112 compared to low-fat diets. High-protein diets decreased (P=0.02) ultrasound MS at d 112 compared to low-protein diets. Carcasses from cattle fed high-fat diets had greater (P=0.03) MS compared to those from cattle fed low-fat diets. No differences (P≥0.14) were observed for HCW, LMA, BF, KPH, or yield grade. These data indicate that final BW was unaffected by protein and fat content of growing calf diets but that increased dietary fat and low dietary protein increased MS.


Structural basis of nectin-1 recognition by pseudorabies virus glycoprotein D.
An early and yet indispensable step in the alphaherpesvirus infection is the engagement of host receptors by the viral envelope glycoprotein D (gD). Of the thus-far identified gD receptors, nectin-1 is likely the most effective in terms of its wide usage by multiple alphaherpesviruses for cell entry. The molecular basis of nectin-1 recognition by the gD protein is therefore an interesting scientific question in the alphaherpesvirus field. Previous studies focused on the herpes simplex virus (HSV) of the Simplexvirus genus, for which both the free gD structure and the gD/nectin-1 complex structure were reported at high resolutions. The structural and functional features of other alphaherpesviral gDs, however, remain poorly characterized. In the current study, we systematically studied the characteristics of nectin-1 binding by the gD of a Varicellovirus genus member, the pseudorabies virus (PRV). We first showed that PRV infects host cells via both human and swine nectin-1, and that its gD exhibits similar binding affinities for nectin-1 of the two species. Furthermore, we demonstrated that removal of the PRV gD membrane-proximal residues could significantly increase its affinity for the receptor binding. The structures of PRV gD in the free and the nectin-1-bound states were then solved, revealing a similar overall 3D fold as well as a homologous nectin-1 binding mode to its HSV counterpart. However, several unique features were observed at the binding interface of PRV gD, enabling the viral ligand to utilize different gD residues (from those of HSV) for nectin-1 engagement. These observed binding characteristics were further verified by the mutagenesis study using the key-residue mutants of nectin-1. The structural and functional data obtained in this study, therefore, provide the basis of receptor recognition by PRV gD.


A Rare Case of Mycotic Aneurysm Due to Methicillin-Sensitive Staphylococcus aureus (MSSA) Bacteremia.
Mycotic aneurysm is defined as an infection of the arterial wall either by fungi or bacteria. Although, a rare complication of infection, it is associated with high morbidity and mortality. We describe a 69-year-old female with a rare thoracic aortic mycotic aneurysm, with no clear source of infection and a predominantly atypical presentation, manifesting primarily as heart failure, at a rural community hospital. Our case also depicts the rapid development of aortitis and mycotic aneurysms. This case highlights the challenges in the diagnosis and management of this condition.


Inhibition of catechol-O-methyltransferase contributes to more stable levodopa plasma levels.
The short plasma half-life limits the antiparkinsonian efficacy of levodopa/carbidopa (LD/CD). Administration of LD/CD with the catechol-O-methyltransferase inhibitor entacapone in one tablet (LCE) may extend plasma half-life of LD and thus its effect on motor symptoms in patients with Parkinson's disease (PD). The objectives of this study were to monitor the motor response to a switch from LD/CD to LCE by a simultaneous performance of an instrumental motor test and rating of motor symptoms and to compare the LD plasma behavior between both conditions in terms of stability. Twenty-one treated PD patients received LD/CD and then the identical oral LD dosage of LCE within a standardized setting on 2 consecutive days. Rating better reflected the motor improvement after LD application than the instrumental test. Motor symptoms of PD patients decreased significantly more during the LCE than the LD/CD condition, probably due to significantly higher LD plasma levels and a significantly less pronounced fall of the LD concentrations following the second LD intake. Our study shows a more stable LD plasma behavior during LCE intake and accordingly a better effect on motor symptoms according to rating outcomes and motor test results to a lesser extent.


Circulating soluble receptor for advanced glycation end products is inversely associated with body mass index and waist/hip ratio in the general population.
Advanced glycation end products, AGEs, and its specific receptor, RAGE, are involved in vascular complications. A role for the soluble form of RAGE (sRAGE), which acts as a decoy for AGE, has been documented in patients with diabetes but no information is available in non-diabetic subjects. The aim of this study was to investigate the association of plasma levels of sRAGE with cardiometabolic risk factors in the general population. In addition we evaluated the relation of the common -374A/T polymorphism of RAGE with plasma levels of sRAGE. One hundred and seventy-six healthy subjects free of diabetes or coronary artery disease untreated for hypertension, dyslipidemia or cardiometabolic related diseases were randomly selected for this study from the general population. Plasma sRAGE were negatively and significantly correlated with BMI, waist/hip circumference ratio and fasting glycemia, while a positive correlation was observed with apolipoprotein A-I. These correlations were observed mainly in women who showed significantly higher sRAGE levels (1744+/-660 pg/mL vs 1414+/-649 pg/mL; P<0.05). In a stepwise regression analysis waist circumference was independently associated with sRAGE and, when waist circumference was excluded, BMI was independently associated with sRAGE. Finally in overweight subjects (BMI>25 kg/m(2)) plasma sRAGE was significantly lower compared to lean subjects (1460+/-640 pg/mL vs 1710+/-693 pg/mL; P<0.05). In healthy subjects plasma levels of sRAGE were negatively correlated with BMI and waist/hip ratio supporting a possible protective role for these proteins before any evidence of diabetic or vascular complications.


Sonography of ovarian fibromas/thecomas.
The sonographic findings in 14 patients with ovarian fibromas/thecomas are described. A broad spectrum of sonographic features are presented and include hypoechoic mass with posterior shadowing (two cases); anechoic mass with good through transmission (with septations, two cases; without septations, four cases), echogenic mass with well-defined posterior wall (three cases); calcified mass (two cases); mixed echogenicity mass (one case). The pattern of a hypoechoic adnexal mass with acoustic shadowing should still suggest a fibroma/thecoma, but in most cases the appearance is nonspecific.


Melting-Freezing Transition of Monolayer Water Confined by Phosphorene Plates.
The two-dimensional material phosphorene has become a focus of the scientific community recently. On the basis of molecular dynamics simulations, we utilize phosphorene as a model material to study the behavior of water molecules confined by two phosphorene plates with nonflat surfaces. As the relative position of the two plates changes, the water molecules first stay in a melting process at 230 K and then exhibit a freezing process. The disparate variations of local confinements induced by the mismatch of the two plates are the key for understanding this extraordinary behavior of water. Our results imply that such nonflat surfaces could be an important factor for understanding or controlling the dynamics of water. The phenomena reported here may enrich the knowledge of water and inspire more applications of similar materials.


Problem size effect in additions and subtractions: an event-related potential study.
The psychophysiological basis of problem size effect in the arithmetical processing of additions and subtractions was studied with event-related brain potentials (ERP). Subjects were presented sequences of seven numbers, and ERPs elicited to the sixth number were analyzed. Two variables were manipulated: operation type (addition and subtraction) and problem size (by adding or subtracting 2, 3 or 4). Results showed two phases in the ERP pattern: an early phase, appearing to reflect automatic processing involved in stimulus identification, and a positive slow wave, believed to be a computing indicator of the subsequent calculation. The amplitude of this positive slow wave was modulated by the problem size (the more problem size, the larger the amplitude), suggesting that the amplitude of this slow wave indexes the activation of the cerebral network underlying problem size effect.


[The mathematical model of the blood circulation and external respiration functional state during high-pressure oxygenation or hyperoxia].
Based on the minimal energy uptake, the proposed mathematical model of blood circulation and external respiration functioning during high-pressure oxygenation or hyperoxia allows solution of the optimization task with restrictions dictated by adequate functioning of the two systems. Optimal levels of oxygen and carbon dioxide pressure in arterial and venous blood, minute blood volume and alveolar ventilation as a function of O2 partial pressure in inspired gas mixture were determined. Calculations are compared with experimentally derived values.


Aldosterone increases cardiac vagal tone via G protein-coupled oestrogen receptor activation.
In addition to acting on mineralocorticoid receptors, aldosterone has been recently shown to activate the G protein-coupled oestrogen receptor (GPER) in vascular cells. In light of the newly identified role for GPER in vagal cardiac control, we examined whether or not aldosterone activates GPER in rat nucleus ambiguus. Aldosterone produced a dose-dependent increase in cytosolic Ca(2+) concentration in retrogradely labelled cardiac vagal neurons of nucleus ambiguus; the response was abolished by pretreatment with the GPER antagonist G-36, but was not affected by the mineralocorticoid receptor antagonists, spironolactone and eplerenone. In Ca(2+)-free saline, the response to aldosterone was insensitive to blockade of the Ca(2+) release from lysosomes, while it was reduced by blocking the Ca(2+) release via ryanodine receptors and abolished by blocking the IP3 receptors. Aldosterone induced Ca(2+) influx via P/Q-type Ca(2+) channels, but not via L-type and N-type Ca(2+) channels. Aldosterone induced depolarization of cardiac vagal neurons of nucleus ambiguus that was sensitive to antagonism of GPER but not of mineralocorticoid receptor. in vivo studies, using telemetric measurement of heart rate, indicate that microinjection of aldosterone into the nucleus ambiguus produced a dose-dependent bradycardia in conscious, freely moving rats. Aldosterone-induced bradycardia was blocked by the GPER antagonist, but not by the mineralocorticoid receptor antagonists. In summary, we report for the first time that aldosterone decreases heart rate by activating GPER in cardiac vagal neurons of nucleus ambiguus.


Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice.
The key mechanism for acetaminophen hepatotoxicity is cytochrome P450 (CYP)-dependent formation of N-acetyl-p-benzoquinone imine, a potent electrophile that forms protein adducts. Previous studies revealed the fundamental role of glutathione, which binds to and detoxifies N-acetyl-p-benzoquinone imine. Glutathione is synthesized from cysteine in the liver, and N-acetylcysteine is used as a sole antidote for acetaminophen poisoning. Here, we evaluated the potential roles of transsulfuration enzymes essential for cysteine biosynthesis, cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH), in acetaminophen hepatotoxicity using hemizygous (Cbs(+/-) or Cth(+/-)) and homozygous (Cth(-/-)) knockout mice. At 4 h after intraperitoneal acetaminophen injection, serum alanine aminotransferase levels were highly elevated in Cth(-/-) mice at 150 mg/kg dose, and also in Cbs(+/-) or Cth(+/-) mice at 250 mg/kg dose, which was associated with characteristic centrilobular hepatocyte oncosis. Hepatic glutathione was depleted while serum malondialdehyde accumulated in acetaminophen-injected Cth(-/-) mice but not wild-type mice, although glutamate-cysteine ligase (composed of catalytic [GCLC] and modifier [GCLM] subunits) became more activated in the livers of Cth(-/-) mice with lower Km values for Cys and Glu. Proteome analysis using fluorescent two-dimensional difference gel electrophoresis revealed 47 differentially expressed proteins after injection of 150 mg acetaminophen/kg into Cth(-/-) mice; the profiles were similar to 1000 mg acetaminophen/kg-treated wild-type mice. The prevalence of Cbs or Cth hemizygosity is estimated to be 1:200-300 population; therefore, the deletion or polymorphism of either transsulfuration gene may underlie idiosyncratic acetaminophen vulnerability along with the differences in Cyp, Gclc, and Gclm gene activities.


Effect of germination on chemical, functional and nutritional characteristics of wheat, brown rice and triticale: a comparative study.
Germination is accompanied by various metabolic reactions in the seed, which lead to alteration of its chemical composition as compared to raw seed. Among the micromolecules, amino acids play an important role in various growth and metabolic activities in seeds. Results indicated that germination altered seeds' chemical composition, which improved/changed the functional properties investigated such as bulk density, foaming capacity, water holding capacity, oil binding capacity and emulsification properties. Essential amino acids were increased during germination, which contributed in enhancing the nutritional quality of protein in seeds. Germination also improved protein-based quality parameters such as essential amino acid index, biological value, protein efficiency ratio and nutritional index in seeds of brown rice, wheat and triticale. Among the grains, the essential amino acid index of brown rice was reported to be highest after germination. Indispensable/essential amino acids such as lysine, methionine, leucine, isoleucine, threonine, phenylalanine and valine showed a significant increase during germination. As a result, amino acid scoring based on the reference pattern of an FAO Expert Consultation Report (2013) for infants and adults was appreciably improved after germination along with an increase in the essential amino acid indices for brown rice (78.78), wheat (76.55) and triticale (73.99). © 2017 Society of Chemical Industry.


Pilchard orthomyxovirus (POMV). I. Characterisation of an emerging virus isolated from pilchards Sardinops sagax and Atlantic salmon Salmo salar.
An orthomyxo-like virus was first isolated in 1998 as an incidental discovery from pilchards Sardinops sagax collected from waters off the South Australian coast. In the following 2 decades, orthomyxo-like viruses have been isolated from healthy pilchards in South Australia and Tasmania. In 2006, an orthomyxo-like virus was also isolated from farmed Atlantic salmon Salmo salar in Tasmania during routine surveillance and, again, from 2012 onwards from diseased Atlantic salmon. Using transmission electron microscopy, these viruses were identified as belonging to the family Orthomyxoviridae. To further characterise the viruses, the genomes of 11 viral isolates were sequenced. The open reading frames (ORFs) that encode 10 putative proteins from 8 viral genome segments were assembled from Illumina MiSeq next generation sequencing (NGS) data. The complete genome of a 2014 isolate was also assembled from NGS, RNA-sequencing (RNA-seq) data, that included conserved motifs that shared commonalities with infectious salmon anaemia virus, rainbow trout orthomyxovirus and Influenzavirus A. The presence of 8 viral proteins translated from genome segments was confirmed by mass spectrometric analysis including 2 novel proteins with no known orthologs. Sequence analysis of the ORFs, non-coding regions and proteins indicated that the viruses had minimal diversity and hence were named pilchard orthomyxovirus (POMV), based on the fish host species of its first isolation. The low homology of POMV proteins with previously characterised orthomyxoviruses suggests that POMV is the first virus to be characterised from a new genus within the Orthomyxoviridae. To facilitate more rapid detection and subsequent diagnostic confirmation of POMV infections, TaqMan and conventional nested PCRs were designed.


Ossification of posterior longitudinal ligaments: evaluation with MRI.
Ossification of the posterior longitudinal ligament is a special subcategory of degenerative disease responsible for compression of the spinal cord. On MR images, T2-weighted sequences are the most effective to evaluate both spinal cord compression due to the ossification and abnormal signal intensity of the cord. Although ossification of the ligaments is well demonstrated on CT and plain radiographs, MRI noninvasively provides useful information about the degree and extent of spinal cord compression, as well as the character of the ossification.


Introduction: food processing and nutritional assimilation in animals.
How animals process and absorb nutrients from their food is a fundamental question in biology. Despite the continuity and interaction between intraoral food processing and post-oesophageal nutritional extraction, these topics have largely been studied separately. At present, we lack a synthesis of how pre- and post-oesophageal mechanisms of food processing shape the ability of various taxa to effectively assimilate nutrients from their diet. The aim of this special issue is to catalyse a unification of these distinct approaches as a functional continuum. We highlight questions that derive from this synthesis, as well as technical advances to address these questions. At present, there is also a skew toward vertebrates in studies of feeding form-function mechanics; by including perspectives from researchers working on both vertebrates and invertebrates, we hope to stimulate integrative and comparative research on food processing and nutritional assimilation. Below, we discuss how the papers in this issue contribute to these goals in three areas: championing a functional-comparative approach, quantifying performance and emphasizing the effects of life history, and food substrate and extrinsic factors in current and future studies of oral food processing and nutritional assimilation. This article is part of the theme issue 'Food processing and nutritional assimilation in animals'.


Development and evaluation of novel surfactant-based elastic vesicular system for ocular delivery of fluconazole.
Fluconazole is a bis-triazole antifungal agent with a low molecular weight (306 Da). It is hydrophilic in nature and has low protein binding. It is available as eye drops for the treatment of ocular mycoses, the second most common cause of blindness in developing countries. However, its administration often results in poor patient compliance and limited use due to its short half-life (15-30 min) and a low log P (0.25). Therefore, fluconazole was incorporated into a novel sorbitan (spans) based elastic (spanlastic) vesicular system with intent to achieve a prolonged and better effect. Spanlastics are to niosomes what Transfersomes(®) are to liposomes. Developed spanlastics consisted of spans and an edge activator prepared by the ether injection method. Developed vesicles were characterized for size, shape, and the number of vesicles/ml by optical microscopy. Entrapment efficiency was determined by the dialysis method, and the ex vivo corneal permeability study was performed using porcine cornea. A 3-tier safety of the novel formulation was established by the Ames test, the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, and in rabbits according to the OECD guidelines 404 and 405. Spanlastics were smaller in size (3 times) and showed a better permeation in comparison to a corresponding niosomal formulation. The system showed an increase (3-fold) in the apparent permeability coefficient compared to the marketed formulation Zocon(®) (0.3% w/v solution of fluconazole) due to its elastic nature. The developed system was found to be stable for 2 months under refrigerated conditions and under extreme storage conditions. Safety was established in terms of genotoxicity (Ames test), cytotoxicity (MTT assay; mouse peritoneal macrophages), acute dermal/eye irritation/corrosion, and chronic eye irritation/corrosion tests (OECD guidelines). The developed system is novel and provides an effective and safe formulation of fluconazole.


The predictive utility of cytokines, procalcitonin and C-reactive protein among febrile pediatric hematology and oncology patients with severe sepsis or septic shock.
Severe sepsis and septic shock are life-threatening for pediatric hematology and oncology patient receiving chemotherapy. Th1/Th2 cytokines, C-reactive protein (CRP), and procalcitonin (PCT) are all thought to be associated with disease severity. The aim of this study was to prospectively verify the utility of Th1/Th2 cytokines and compare them with PCT and CRP in the prediction of adverse outcomes. Data on patients were collected from January 1, 2011, to December 31, 2020. Blood samples were taken for Th1/Th2 cytokine, CRP, and PCT measurements at the initial onset of infection. Severe infection (SI) was defined as severe sepsis or septic shock. Th1/Th2 cytokine levels were determined by using flow cytometric bead array technology. In total, 7,735 febrile episodes were included in this study. For SI prediction, the AUCs of IL-6, IL-10 and TNF-α were 0.814, 0.805 and 0.624, respectively, while IL-6 and IL-10 had high sensitivity and specificity. IL-6 > 220.85 pg/ml and IL-10 > 29.95 pg/ml had high odds ratio (OR) values of approximately 3.5 in the logistic regression. Within the subgroup analysis, for bloodstream infection (BSI) prediction, the AUCs of IL-10 and TNF-α were 0.757 and 0.694, respectively. For multiorgan dysfunction syndrome (MODS) prediction, the AUC of CRP was 0.606. The AUC of PCT for mortality prediction was 0.620. In conclusion, IL-6 and IL-10 provide good predictive value for the diagnosis of SI. For children with SI, IL-10 and TNF-α are associated with BSI, while CRP and PCT are associated with MODS and death, respectively.


Neurotoxicity of benzylpenicillin in experimental Escherichia coli meningitis.
The neurotoxic potential of benzylpenicillin administered intravenously as a continuous infusion was studied in rabbits with experimental Escherichia coli meningitis. As controls a group of rabbits was injected with saline into the cisterna magna. The concentrations of benzylpenicillin in serum, CSF and brain tissue fluid were studied at onset of epileptogenic electroencephalographic activity (thirteen rabbits) or convulsions (ten rabbits), with a previously developed method for neurotoxicity studies. E. coli meningitis did not increase the neurotoxicity of benzylpenicillin, despite high concentrations of the drug in both CSF and brain tissue fluid. The intracisternal injection of saline in the control group produced slight pleocytosis in some rabbits indicating some degree of damage of the blood-CSF barrier.


[Role of incretins in energy metabolism and weight changes after smoking cessation].
Incretine-based therapies are frequently used to treat patients with type 2 diabetes. Some of these drugs are used to induce weight loss. Liraglutide, a glucagon-like peptide 1 receptor agonist, was recently approved for the treatment of obesity. Smoking cessation is associated with weight gain. The mechanism responsible for the increase in body weight post cessation remains unclear. While increased caloric intake may play a role, weight gain may also be linked to nicotine, which has been shown to stimulate smokers basal metabolic rate. The effect of incretines on body weight and energy metabolism after smoking cessation is unclear. Recently published data suggests that incretine hormones may not be involved in the above mentioned changes.


Mesoporous silica nanoparticles incorporated hybrid monolithic stationary phase immobilized with pepsin for enantioseparation by capillary electrochromatography.
In this study, a novel mesoporous silica nanoparticles incorporated chiral hybrid monolithic stationary phase was developed. The stationary phase was firstly prepared by an in situ copolymerization of amino-modified mesoporous silica nanoparticles (NH2-MSN), glycidyl methacrylate (GMA), and ethylene dimethacrylate (EDMA) and then functionalized with pepsin as chiral selector. The incorporated mesoporous silica nanoparticles provided additional interactions sites, and in turn yielded different enantioselectivity thus enhancing the overall separation. The column was successfully employed for enantioseparation of fifteen basic chiral drugs in capillary electrochromatography. Effects of nanoparticles percentage, pepsin concentration, the pH of running buffer and the applied voltage were investigated. All the analytes could be eluted in less than ten minutes and nine of them could achieve baseline separation. Satisfactory repeatabilities with relative standard deviations less than 4.2% were achieved through intraday, interday, column-to-column and batch-to-batch investigations. These results indicated that the simultaneous utilization of the unique properties of mesoporous silica nanoparticles and versatile features of monoliths could be a promising strategy for enantioseparation.


[Emerging disease-modifying therapies for multiple sclerosis].
Multiple sclerosis(MS) is an inflammatory demyelinating disease of the central nervous system with yet undetermined etiology. The incidence of MS is rapidly rising in Japan, and in addition to four already-approved disease-modifying therapies (DMTs), namely interferon β1b, interferonβ1a, fingolimod and natalizumab, several new DMTs are currently under clinical trials. In this article, emerging DMTs such as alemtuzumab, dimethyl fumarate and teriflunomide as well as fampridine, a novel symptomatic treatment for gait disturbance, and anti-LINGO-1 monoclonal antibody, a possible candidate for future remyelination therapy, will be reviewed. Future treatment strategies such as induction therapy and combined therapy for MS will also be discussed.


Autoimmunity induced by chemicals.
Immunotoxicologic studies have demonstrated that autoimmune responses and/or autoimmune diseases are induced in humans and experimental animals by chronic exposure to various chemicals. The present review is focused on seven groups of chemically induced human disorders, i.e. systemic lupus erythematosus, autoimmune hemolytic anemia, myasthenia gravis, pemphigus, glomerulonephritis, thyroiditis and hepatitis. Results obtained from studies of the available experimental counterparts of these diseases, i.e. those models obtained from the exposure of laboratory animals to various chemicals, are then analyzed. Finally, we present the lessons that can be derived from immunotoxicologic investigations regarding mechanisms of induction, heterogeneity of chemicals involved, humoral vs. cellular immune responses and genetic predisposition to chemically induced autoimmunity.


A Fast and Reliable Real-Time PCR Method for Detection of Ten Animal Species in Meat Products.
Species substitution in meat products is a common problem reported worldwide. This type of food fraud is, typically, an intentional act for economic gain, using sources of low-priced meats in high-value meat products. Consequences include economic, health, and religious concerns. Highly sensitive and efficient techniques are thus required to detect meat species. This paper describes a method based on real-time PCR to detect 10 animal species (Bos taurus, Sus scrofa, Ovis aries, Capra hircus, Gallus gallus, Meleagris gallopavo, Bubalus bubalis, Equus caballus, Felis catus, and Canis familiaris) in meat product. The method combines species-specific and universal (used here as internal positive control) primers, and applies melt curve analysis for amplicon checking. Method accuracy was evaluated on 46 experimental meat mixtures and all species were correctly identified in all cases, at 1% test sensitivity. Analysis of 14 commercial meat products revealed that 6 of 14 samples had nondeclared bovine and/or chicken material. We performed an interlaboratory comparison using the reference meat mixtures and commercial samples, achieving 100% of reproducibility. The developed test proved to be effective and reliable for routine analysis of meat products. This paper describes a fast and reliable method for species detection in meat products based on real-time PCR. It can be applied for analysis of in natura or processed meat. The method proposed here can play an important role in controlling the origin of meat products, ensuring their quality and safety for the entire food industry-producers to consumers.


Modeling transfer of Escherichia coli O157:H7 and Listeria monocytogenes during preparation of fresh-cut salads: impact of cutting and shredding practices.
Cutting and shredding of leafy vegetables increases the risk of cross contamination in household settings. The distribution of Escherichia coli O157:H7 and Listeria monocytogenes transfer rates (Tr) between cutting knives and lettuce leaves was investigated and a semi-mechanistic model describing the bacterial transfer during consecutive cuts of leafy vegetables was developed. For both pathogens the distribution of log10Trs from lettuce to knife was towards low values. Conversely log10Trs from knife to lettuce ranged from -2.1 to -0.1 for E. coli O157:H7 and -2.0 to 0 for L. monocytogenes, and indicated a more variable phenomenon. Regarding consecutive cuts, a rapid initial transfer was followed by an asymptotic tail at low populations moving to lettuce or residing on knife. E. coli O157:H7 was transferred at slower rates than L. monocytogenes. These trends were sufficiently described by the transfer-model, with RMSE values of 0.426-0.613 and 0.531-0.908 for L. monocytogenes and E. coli O157:H7, respectively. The model showed good performance in validation trials but underestimated bacterial transfer during extrapolation experiments. The results of the study can provide information regarding cross contamination events in a common household. The constructed model could be a useful tool for the risk-assessment during preparation of leafy-green salads.


Dissolved organic carbon in glaciers of the southeastern Tibetan Plateau: Insights into concentrations and possible sources.
Dissolved organic carbon (DOC) released from glaciers has an important role in the biogeochemistry of glacial ecosystems. This study focuses on DOC from glaciers of the southeastern Tibetan Plateau, where glaciers are experiencing rapid shrinkage. We found that concentrations of DOC in snowpits (0.16±0.054 μg g-1), aged snow (0.16±0.048 μg g-1), and bare ice (0.18±0.082 μg g-1) were similar across the southeastern Tibetan Plateau, but were slightly lower than those in other glaciers on the Tibetan Plateau. Vertical variations of DOC, particulate organic carbon, black carbon, and total nitrate in snowpit showed no systematic variations in the studied glaciers, with high values of DOC occurring in the ice or dusty layers. We estimated the export of DOC and particulate organic carbon from glaciers to be 1.96±0.66 Gg yr-1 and 5.88±2.15 Gg yr-1 in this region, respectively, indicating that organic carbon released from glacier meltwater may be affecting downstream ecosystems. Potential sources of the air masses arriving at the southeastern Tibetan glaciers include South Asia, Central Asia, Middle East, and northwest China. Emissions from biomass burning of South Asia played an important role in the deposition of DOC to the glacier, which can be evidenced by backward trajectories and fire spot distributions from MODIS and CALIPSO images. Our findings suggest that anthropogenic aerosols contribute abundant DOC to glaciers on the southeastern Tibetan Plateau. The pronounced rate of glacial melting in the region may be delivering increased quantities of relic DOC to downstream rivers.


Stroke onset and rehabilitation: time lag as a factor in treatment outcome.
Increasing time between stroke onset and initiation of rehabilitation has been associated with poorer outcome in published reports. However, this conclusion is difficult to generalize because of variations in study sample size and selection. The present study examines rehabilitation outcomes for 687 patients admitted to a private rehabilitation center. Outcome variables include measures of ambulation, transfers, upper extremity function, and self-care in dressing, feeding, hygiene, communication, and household and homemaking activities. The association of outcome with fourteen variables, including time since onset, is explored using a multiple regression approach. Results suggest that time since stroke onset before admission to a rehabilitation facility has a minor effect on outcome when adjustments for other variables are taken into account. Its effects appear to be limited to treatment received in physical therapy, such as transfers and ambulation training. Other variables such as patient age, judgment, attention to task, and skill carryover are consistently associated with outcome measures and should be considered when assessing rehabilitation potential.


Priapism in teenage boys following depot testosterone.
Priapism is rare in children and may result in erectile dysfunction and sexual aversion behaviours. Testosterone therapy is commonly regarded as safe in children and is widely used in constitutional delay of growth and puberty, hypogonadism, hypospadias and micropenis. We report two cases of priapism in teenage boys with constitutional delay of growth and puberty after a change in the formulation of depot testosterone. One case required surgical intervention and the other was preceded by stuttering priapism. These cases illustrate the importance of patient and/or parent counselling before testosterone administration and consideration of lower doses in at-risk patients.


Pressure-induced structural transformations and new polymorphs in BiVO4.
BiVO4 has attracted much attention in recent years due to its active photocatalytic and microwave dielectric properties. BiVO4 exhibits a rich structural polymorphism, and its properties strongly depend on the crystalline phase. Therefore, it is of great importance to achieve an easy control of its crystalline phase. In the present work, phase stability and vibrational properties of fergusonite- and zircon-type BiVO4 are investigated up to 41.6 GPa by in situ synchrotron X-ray diffraction (XRD), Raman spectroscopy, and first principles calculation. Upon compression, although having different initial structures, both types of BiVO4 consecutively transform to scheelite- and β-fergusonite structures. For the first time reported for BiVO4, the β-fergusonite structure is determined using first principles computational techniques and from refinement of the XRD data. Along the way, one new phase of BiVO4 is theoretically predicted at higher pressures. Moreover, both the fergusonite-to-scheelite and scheelite-to-β-fergusonite transitions are reversible, while the zircon-to-scheelite transition is irreversible. A large volume collapse is observed associated with each phase transition, and the equations of state for different types of BiVO4 have been determined. These results provide new insights into the relationship between different structural types in the AVO4 family.


Influence of nitric oxide on the activity of cuneate neurons in the rat.
Some neurons of main and external cuneate nuclei are immunoreactive for nitric oxide (NO) synthase, suggesting a role for endogenous NO in the early stages of somatosensory processing. We tested this hypothesis by investigating the possibility that NO modulates cuneate discharge. We observed that both spontaneous and N-methyl-D-aspartate-evoked activities of cuneate neurons were decreased by NO precursor L-arginine. The inhibition of NO synthase, by application of N-nitro-L-arginine methyl ester, instead, abolished the depressant effect induced by L-arginine. Our data suggest a NO modulation of cuneate neurons and provide support for a physiologic role not only in increasing the signal-to-noise ratio in the excited cells but also in a form of surround inhibition.