3-Amino-1-hydroxypropylidine-1-bisphosphonate (AHPrBP) suppresses not only the induction of new, but also the persistence of existing bone-forming surfaces in rat cancellous bone.
Although bone formation is coupled to resorption in both man and the rat, it is not known whether similar mechanisms are involved in these two species. To investigate this, we have attempted to further characterise the suppression of bone formation by the potent bone resorption inhibitor 3-amino-1-hydroxypropylidine-1-bisphosphonate (AHPrBP) in ovariectomised adult female rats. To see whether AHPrBP affects the activity of preexisting bone forming surfaces, as opposed to preventing the induction of new forming surfaces, three consecutive fluorochrome labels were administered 3, 10, and 17 days prior to sacrifice. The percentage of trabecular surface covered by first and second labels only (arrested surface), second and third labels only (induction surface), and all three labels (persisting surface) were recorded at the secondary spongiosa of the proximal tibial metaphysis. As expected, ovariectomy increased the proportion of induction surfaces, with AHPrBP preventing this increase. However, AHPrBP also reduced the proportion of persisting surfaces, implying inhibition of the activity of preexisting bone-forming surfaces. We conclude that resorption inhibitors such as AHPrBP inhibit bone formation in the rat in a manner which appears inconsistent with the site-specific coupling seen in the adult human skeleton.


Pan-neurotrophin receptor p75NTR expression is strongly induced in lesional atopic mast cells.
Neurotrophins such as nerve growth factor or brain-derived neurotrophic factor influence neuronal proliferation and differentiation via the low-affinity pan-neurotrophin receptor p75NTR that may play a pivotal role in linking the immune with the nervous system. Because the precise regulation of p75NTR gene transcription in mast cells under states of allergic inflammation has not been investigated in detail so far, the present studies assessed the gene regulation and expression of this receptor. Transcriptional expression of p75NTR in human skin was studied in isolated cutaneous cells by means of RT-PCR. In situ lesional mast cell p75NTR expression was analysed by immunohistochemistry. The p75NTR mRNA expression was found in isolated human skin mast cells and keratinocytes. Lower mRNA levels were present in fibroblasts and melanocytes but no transcripts were found in endothelial cells. The p75NTR protein expression was found in situ in lesional and non-lesional mast cells. A significantly increased expression of p75NTR protein was found in atopic dermatitis lesional mast cells when compared with control mast cell expression (P<0.05). The demonstration of an increased level of p75NTR gene transcription in lesional mast cells points to an induction of low-affinity neurotrophin receptor sensitivity of mast cells under states of allergic inflammation. Topically administered neurotrophin receptor-modulating compounds may act as anti-inflammatory mediators in cutaneous allergic inflammation.


Interventional approaches to mitral regurgitation. What's in the pipeline?
Mitral regurgitation (MR) is the most prevalent valvular heart disease in the Western world. Surgical repair is the gold standard for the treatment of degenerative MR in low-risk patients. Given the rising number of patients with functional MR and higher operative risk due to relevant comorbidities and increasing age, interventional approaches to repair or replace diseased mitral valves are on the rise. However, the complex anatomy and physiology of the mitral valve and its adjacent valve apparatus bear major challenges. To date, only the MitraClip device has been used in a large number of patients; however, several other devices and systems specifically targeted at different underlying pathologies of MR are currently under development. In addition to valve repair, the first steps toward mitral valve replacement have been taken. The present article reviews the current state of the art of interventional approaches to mitral valve disease and its future perspectives.


A first dataset toward a standardized community-driven global mapping of the human immunopeptidome.
We present the first standardized HLA peptidomics dataset generated by the immunopeptidomics community. The dataset is composed of native HLA class I peptides as well as synthetic HLA class II peptides that were acquired in data-dependent acquisition mode using multiple types of mass spectrometers. All laboratories used the spiked-in landmark iRT peptides for retention time normalization and data analysis. The mass spectrometric data were deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD001872. The generated data were used to build HLA allele-specific peptide spectral and assay libraries, which were stored in the SWATHAtlas database. Data presented here are described in more detail in the original eLife article entitled 'An open-source computational and data resource to analyze digital maps of immunopeptidomes'.


Lack of effects of isosorbide-5-mononitrate on hepatic hemodynamics in HBsAg-positive cirrhosis.
We conducted a randomized controlled hemodynamic study to evaluate the effect of placebo and 20 mg isosorbide-5-mononitrate, a long-acting organic nitrate, in 19 patients with HBsAg-positive cirrhosis by the simultaneous measurement of portal venous pressure and wedged hepatic venous pressure. Baseline values for the two groups were similar. One hour after oral administration of 20 mg isosorbide-5-mononitrate in 10 patients, mean arterial pressure, mean pulmonary arterial pressure and pulmonary capillary wedge pressure significantly decreased from 92 +/- 13 (mean +/- S.D.) to 82 +/- 14 mmHg, from 12.9 +/- 4.5 to 9.3 +/- 2.4 mmHg and from 6.9 +/- 3.4 to 4.3 +/- 1.8 mmHg, respectively. However, both portal venous pressure gradient (from 18.1 +/- 3.6 to 17.5 +/- 3.0 mmHg) and hepatic venous pressure gradient (from 17.8 +/- 5.2 to 16.6 +/- 5.3 mmHg) remained unchanged during the study. In six patients who received 20 mg isosorbide-5-mononitrate twice daily for 7 days, hepatic venous pressure gradient remained unaltered as compared to basal and 1-hr values. There was no significant change in cardiac index, heart rate or systemic vascular resistance in either immediate (1-hr) or delayed (7-day) studies. Three patients (30%) developed mild headache or dizziness and two patients (20%) demonstrated systolic hypotension (less than mmHg) during the immediate study. This study shows that isosorbide-5-mononitrate appears to have no effect in treating portal hypertension in patients with HBsAg-positive cirrhosis. In addition, the isosorbide-5-mononitrate may affect the systemic circulation more than the portal circulation.


Misleading effects of impurities derived from the extremely long-lived electron-transfer state of 9-mesityl-10-methylacridinium ion.
9-Mesityl-10-methylacridinium ion has an extremely long-lived electron-transfer state, which results in the misleading effects of electron donor impurities contained in benzonitrile to produce the stable acridinyl radical.


Tissue characterization of brain tumors during and after pion radiation therapy.
Negative Pi-mesons (pions) are applied at the Paul Scherrer Institute in the radiotherapy of highly malignant gliomas using a dose escalation program. The therapy effects of 7 randomly selected patients were followed up by 62 MRI examinations. The quantification of the effects is based on the relaxation times T1 and T2, which are acquired by a new designed multi-echo multiple saturation recovery imaging technique. As a summary of the results, roughly two reaction types are observed. For both types the relaxation times increase up to two to three months after the radiation therapy. Then in one type (two patients) the T1 and T2 values of the tumors, and of the edemas surrounding the tumors, further increase, indicating an unfavorable prognosis. In the other type (five patients) the relaxation times drop down towards, or even below, their initial values, reflecting the onset of the reparation processes in the tissue. This later behaviour reflects an at least temporary control of the disease; that is, the short term prognosis for these patients is more favorable. It further can be concluded, with respect to our MR parameters, that the radiotolerance of healthy brain tissue is much higher than that of malignant glioma tissue, despite the fact that these tumors are very seldom definitively radiosensible.


Structure-function relationships using spectral-domain optical coherence tomography: comparison with scanning laser polarimetry.
To evaluate and compare the regional relationships between visual field sensitivity and retinal nerve fiber layer (RNFL) thickness as measured by spectral-domain optical coherence tomography (OCT) and scanning laser polarimetry. Prospective cross-sectional study. One hundred and twenty eyes of 120 patients (40 with healthy eyes, 40 with suspected glaucoma, and 40 with glaucoma) were tested on Cirrus-OCT, GDx VCC, and standard automated perimetry. Raw data on RNFL thickness were extracted for 256 peripapillary sectors of 1.40625 degrees each for the OCT measurement ellipse and 64 peripapillary sectors of 5.625 degrees each for the GDx VCC measurement ellipse. Correlations between peripapillary RNFL thickness in 6 sectors and visual field sensitivity in the 6 corresponding areas were evaluated using linear and logarithmic regression analysis. Receiver operating curve areas were calculated for each instrument. With spectral-domain OCT, the correlations (r(2)) between RNFL thickness and visual field sensitivity ranged from 0.082 (nasal RNFL and corresponding visual field area, linear regression) to 0.726 (supratemporal RNFL and corresponding visual field area, logarithmic regression). By comparison, with GDx-VCC, the correlations ranged from 0.062 (temporal RNFL and corresponding visual field area, linear regression) to 0.362 (supratemporal RNFL and corresponding visual field area, logarithmic regression). In pairwise comparisons, these structure-function correlations were generally stronger with spectral-domain OCT than with GDx VCC and with logarithmic regression than with linear regression. The largest areas under the receiver operating curve were seen for OCT superior thickness (0.963 ± 0.022; P < .001) in eyes with glaucoma and for OCT average thickness (0.888 ± 0.072; P < .001) in eyes with suspected glaucoma. The structure-function relationship was significantly stronger with spectral-domain OCT than with scanning laser polarimetry, and was better expressed logarithmically than linearly. Measurements with these 2 instruments should not be considered to be interchangeable.


Enzymic programs of rat bone marrow and the impact of acivicin and tiazofurin.
The in vivo actions of two antimetabolites, acivicin (NSC-163501) and tiazofurin (NSC-286193), were examined on the enzymic programs of rat bone marrow. From the bone marrow of the femurs, 100,000 g supernatant fractions were prepared; enzymic activities were measured by isotopic assays, and cellularity was determined. In the normal bone marrow, the specific activities of pyrimidine de novo synthetic enzymes, CDP reductase, dTMP synthase, CTP synthase, carbamoyl-phosphate synthase II (synthase II), orotidine 5'-phosphate decarboxylase and aspartate carbamoyltransferase, were 1, 2.7, 5, 10, 63 and 601 nmol/hr/mg protein, respectively, whereas those of the salvage enzymes, deoxycytidine, thymidine, cytidine and uridine kinases were 3, 43, 149, and 367 nmol/hr/mg protein, respectively. In purine biosynthesis, the activities of the de novo synthetic enzymes, IMP dehydrogenase, formylglycinamidine ribonucleotide (FGAM) synthase, GMP synthase, amidophosphoribosyl-transferase (AT) and adenylosuccinate synthase were 16, 8, 107, 78 and 124 nmol/hr/mg protein, respectively, and those of the salvage enzymes, adenine, hypoxanthine and guanine phosphoribosyl-transferases, were 340, 407, and 1018 nmol/hr/mg protein, respectively. The sequence of events was elucidated after a single i.p. injection of acivicin (5 mg/kg) or tiazofurin (200 mg/kg). Within 2 hr after acivicin injection, CTP, GMP and FGAM synthases lost 85-90%, while AT and synthase II lost 50 and 80%, respectively, of their activities. The activities rose to near normal range by 72-96 hr. The bone marrow cellularity decreased, reaching a nadir at 24 and 48 hr, and returning to normal range by 72 and 92 hr; thymidine kinase activity followed a similar pattern. Tiazofurin injection depressed IMP dehydrogenase activity to 20% by 2 hr with a rebound to normal range by 48 and 72 hr. The cellularity decreased more slowly, reaching its lowest point at 24 hr and returning to normal range at 72 hr. For acivicin the marked depletion of the activities of the glutamine-utilizing enzymes and for tiazofurin that of IMP dehydrogenase might account, in part at least, for the bone marrow toxicity of these antimetabolites. Because of the presence in the bone marrow of high activities of purine and pyrimidine salvage enzymes, it should be possible to design methods utilizing nucleosides and nucleobases to protect the bone marrow from the action of antimetabolites.


Noise reduction using novel loss functions to compute tissue mineral density and trabecular bone volume fraction on low resolution QCT.
Micro-structural parameters of the thoracic or lumbar spine generally carry insufficient accuracy and precision for clinical in vivo studies when assessed on quantitative computed tomography (QCT). We propose a 3D convolutional neural network with specific loss functions for QCT noise reduction to compute micro-structural parameters such as tissue mineral density (TMD) and bone volume ratio (BV/TV) with significantly higher accuracy than using no or standard noise reduction filters. The vertebra-phantom study contained high resolution peripheral and clinical CT scans with simulated in vivo CT noise and nine repetitions of three different tube currents (100, 250 and 360 mAs). Five-fold cross validation was performed on 20466 purely spongy pairs of noisy and ground-truth patches. Comparison of training and test errors revealed high robustness against over-fitting. While not showing effects for the assessment of BMD and voxel-wise densities, the filter improved thoroughly the computation of TMD and BV/TV with respect to the unfiltered data. Root-mean-square and accuracy errors of low resolution TMD and BV/TV decreased to less than 17% of the initial values. Furthermore filtered low resolution scans revealed still more TMD- and BV/TV-relevant information than high resolution CT scans, either unfiltered or filtered with two state-of-the-art standard denoising methods. The proposed architecture is threshold and rotational invariant, applicable on a wide range of image resolutions at once, and likely serves for an accurate computation of further micro-structural parameters. Furthermore, it is less prone for over-fitting than neural networks that compute structural parameters directly. In conclusion, the method is potentially important for the diagnosis of osteoporosis and other bone diseases since it allows to assess relevant 3D micro-structural information from standard low exposure CT protocols such as 100 mAs and 120 kVp.


Adult male report of childhood sexual abuse by mothers: case descriptions, motivations and long-term consequences.
Sexual abuse of male children by their mothers is rarely reported. However, it may not be as rare as commonly believed. Eight case histories are presented which exemplify mothers' sexual abuse of their sons to satisfy the mothers' own needs. In no case was the mother psychotic. In seven cases the mother began the seduction and sexual abuse while the son was prepubescent and continued until at least early adolescence. As adults, each of these sons experienced difficulty maintaining an intimate emotional and sexual relationship with one person, and most presented with some degree of depression. Substance abuse was present in five of eight cases.


Synthesis, Structural Characterization and Coordination Chemistry of a NHC-stabilized Iminoborane.
NHC-stabilized iminoborane 4 was prepared and structurally characterized via the reduction of a cationic iminoborane with potassium graphite. Compound 4 could be served as a supporting ligand for the synthesis of both main group and transition metal complexes through variable coordination modes. This study demonstrates the rich coordination chemistry of the Lewis base-stabilized iminoborane.


Coordination abilities of piperyd-1-yl-methane-1,1-diphosphonic acids towards zinc(II), magnesium(II) and calcium(II): potentiometric and NMR studies.
The combination of the pH-metric and NMR studies is used to examine the stabilities and coordination modes as well as related structural aspects of zinc(II), magnesium(II) and calcium(II) complexation to piperyd-1-yl-methane-1,1-diphosphonic acid (1) and its derivatives containing a topologically modified piperidine ring (2-7). The studied compounds coordinate metal ions exclusively via the phosphonate functions with a nitrogen atom remaining protonated over the whole range of studied pH. Compounds 1-6 readily form soluble multinuclear complexes of type [M(3)(HL)(2)] and [M(3)(HL)(3)](3-) with Zn(2+) or [M(2)(H(2)L)(2)] with Ca(2+) and Mg(2+). These species are formed based on dimers consisting of two head-to-head arranged molecules linked by strong symmetrical hydrogen bonds. The placement of the two methyl groups at 2- and 6-positions on the piperidine ring precludes the molecular recognition via similar hydrogen bonds and accounts for different complexation properties of 7 compared to 1-6. The role that the metal coordination plays on conformation dynamics in 1-7 is also discussed.


Influence of ions on the coagulation and removal of fullerene in aqueous phase.
Increased attention has been paid to the transportation and removal efficiencies of nanoparticles during water treatment. Here, C₆₀ was selected as nano-pollutant to investigate its removal efficiency in the process of coagulation by Al(3+) in regard to different ions. Negatively charged C₆₀ tends to adsorb hydrated Al(3+) to form positively charged C₆₀-Al(OH)(n)((m-δ)+) particles, leading to the restabilization of C₆₀. Multivalent anions, i.e., CO₃(2-), SO₄(2-), HPO₄(2-),and humic acid (HA), were shown to bridge the C₆₀-Al (OH)(n)((m-δ)+) particles together and induce the formation of flocs, resulting in enhanced C₆₀ removal (>80%). Monovalent anions, such as OH(-), NO₃(-) and the surfactant SDS, were not able to bridge the C₆₀-Al (OH)(n)((m-δ)+) particles together; hence, they had no effect on the C₆₀ removal. The addition of Na(+) and Ca(2+) increased the ionic strength of the aqueous solution, which compressed the electric double layer and induced the C₆₀-Al(OH)(n)((m-δ)+) particles to aggregate. However, they are not the essential factors for flocs formation. This is the first study to report the bridging phenomena between multivalent anions and C₆₀-Al(OH)(n)((m-δ)+) in water treatments, and it highlights that the presence of multivalent anions and cations in raw water are important for the effective removal of C₆₀.


Interleukin-33 is released in spinal cord and suppresses experimental autoimmune encephalomyelitis in mice.
Interleukin-33 (IL-33) is usually expressed in the nucleus as a non-histone chromatin-associated protein. After passively released by necrotic cells, it functions as an IL-1 family member. IL-33 is highly expressed in the central nervous system (CNS), whether IL-33 is actively released in the CNS and involved in experimental autoimmune encephalomyelitis (EAE) remains unclear. In this study, we found that IL-33 and receptor ST2 were expressed in the spinal cord of naïve mice. Compared to naive situation, the intracellular IL-33 was dramatically decreased and extracellular IL-33 was markedly increased in the spinal cord in the pre-onset, onset and peak stage of EAE. In the chronic stage, the reverse happened. The decrease of intracellular IL-33 was related to the activation of astrocytes and the damage of neurons in situ during EAE. Astrocytes secreted IL-33 actively upon inflammatory stimulation in vitro. Furthermore, blockage of the CNS-derived IL-33 exacerbated EAE development. Our data demonstrated that IL-33 was released by activated astrocytes actively, and by damaged neurons during EAE. It plays a suppressive role in EAE development via an autocrine or paracrine manner. Our findings are helpful to understand the release feature and function of the CNS-derived IL-33 and supply a potential therapeutic target for multiple sclerosis.


Comparison of [14C]FMAU, [3H]FEAU, [14C]FIAU, and [3H]PCV for monitoring reporter gene expression of wild type and mutant herpes simplex virus type 1 thymidine kinase in cell culture.
To assess the optimal reporter probe/reporter gene combination for monitoring herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene expression, we compared the cellular uptake of 1-(2'-fluoro-2'-deoxy-D-arabinofuranosyl)-5-methyluracil (FMAU), 2'-fluoro-2'-deoxyarabinofuranosyl-5-ethyluracil (FEAU), 2'-fluoro-2'-deoxy-beta-D-arabinofuranosyl-5-iodouracil (FIAU) and penciclovir (PCV) in both HSV1-tk and HSV1-sr39tk expressing cells. For stably transfected cell studies, C6 rat glioma cells, C6 HSV1-tk transfectant, C6 mutant HSV1-sr39tk transfectant, rat Morris hepatoma cells (MH3924A), and MH3924A HSV1-tk transfectant cells were used. For adenoviral infection studies, C6 rat glioma cells were exposed to serial titers of AdCMV-HSV1-tk, AdCMV-HSV1-sr39tk, or AdCMV-fluc for 24 hours. These cells were incubated with [(14)C]FMAU, [(3)H]FEAU, [(14)C]FIAU, and [(3)H]PCV, and cellular uptake of radioactivity was measured. [(3)H]FEAU exhibited the highest or second highest accumulation and the most selectivity regardless of the mode of gene transfer for both HSV1-tk and mutant HSV1-sr39tk reporter genes. This combination of high accumulation and high selectivity for both HSV1-tk and HSV1-sr39tk makes suitably radiolabeled FEAU a promising candidate as a radiotracer for imaging HSV1-tk/HSV1-sr39tk gene expression in living subjects.


Cyclophilin A modulates the sensitivity of HIV-1 to host restriction factors.
Many mammalian species express restriction factors that confer host resistance to retroviral infection. Here we show that HIV-1 sensitivity to restriction factors is modulated by cyclophilin A (CypA), a host cell protein that binds the HIV-1 capsid protein (CA). In certain nonhuman primate cells, the CA-CypA interaction is essential for restriction: HIV-1 infectivity is increased >100-fold by cyclosporin A (CsA), a competitive inhibitor of the interaction, or by an HIV-1 CA mutation that disrupts CypA binding. Conversely, disruption of CA-CypA interaction in human cells reveals that CypA protects HIV-1 from the Ref-1 restriction factor. These findings suggest that HIV-1 has co-opted a host cell protein to counteract restriction factors expressed by human cells and that this adaptation can confer sensitivity to restriction in unnatural hosts. Manipulation of HIV-1 CA recognition by restriction factors promises to advance animal models and new therapeutic strategies for HIV-1 and AIDS.


Serotonin in the development of pituitary-adrenocortical response to stress.
Adrenocortical response to intraperitoneal 5-HT administration and immobilization stress is observed in rats as early as the 4th day of life when response to intraventricular administration is still absent. Dexamethasone pretreatment influences plasma corticosteroid response to stimulation of peripheral 5-HT receptors in adult rats but does not alter such response to intraperitoneal 5-HT administration and immobilization in 4-day-old rats. Plasma 11-OHCS response to this stress increases considerably by days 12-16, when stress-induced changes in brain 5-HIAA are also observed. The increase of adrenocortical response to stress coincides with the appearance of brain 5-HT metabolic stress-induced changes as well as pituitary-adrenocortical response to centrally administrators, rather than to brain 5-HT, the latter appearing to become essential for the pituitary-adrenocortical system response to this stress from days 12-16 of life.


The novel HLA-DQB1*03:499N allele identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.
HLA-DQB1*03:499N differs from HLA-DQB1*03:01:01:01 by one nucleotide in exon 2.


Tissue specificity of oncogenic BRAF targeted to lung and thyroid through a shared lineage factor.
Cells of origin in cancer determine tumor phenotypes, but whether lineage-defining transcription factors might influence tissue specificity of tumorigenesis among organs with similar developmental traits are unknown. We demonstrate here that tumor development and progression markedly differ in lung and thyroid targeted by Braf mutation in Nkx2.1CreERT2 mice heterozygous for Nkx2-1. In absence of tamoxifen, non-induced Nkx2.1CreERT2;BrafCA/+ mutants developed multiple full-blown lung adenocarcinomas with a latency of 1-3 months whereas thyroid tumors were rare and constrained, although minute BrafCA activation documented by variant allele sequencing was similar in both tissues. Induced oncogene activation accelerated neoplastic growth only in the lungs. By contrast, NKX2-1+ progenitor cells were equally responsive to constitutive expression of mutant Braf during lung and thyroid development. Both lung and thyroid cells transiently downregulated NKX2-1 in early tumor stages. These results indicate that BRAFV600E-induced tumorigenesis obey organ-specific traits that might be differentially modified by a shared lineage factor.


Development and validation of a novel risk prediction algorithm to estimate 10-year risk of oesophageal cancer in primary care: prospective cohort study and evaluation of performance against two other risk prediction models.
Methods to identify patients at increased risk of oesophageal cancer are needed to better identify those for targeted screening. We aimed to derive and validate novel risk prediction algorithms (CanPredict) to estimate the 10-year risk of oesophageal cancer and evaluate performance against two other risk prediction models. Prospective open cohort study using routinely collected data from 1804 QResearch® general practices. We used 1354 practices (12.9 M patients) to develop the algorithm. We validated the algorithm in 450 separate practices from QResearch (4.12 M patients) and 355 Clinical Practice Research Datalink (CPRD) practices (2.53 M patients). The primary outcome was an incident diagnosis of oesophageal cancer found in GP, mortality, hospital, or cancer registry data. Patients were aged 25-84 years and free of oesophageal cancer at baseline. Cox proportional hazards models were used with prediction selection to derive risk equations. Risk factors included age, ethnicity, Townsend deprivation score, body mass index (BMI), smoking, alcohol, family history, relevant co-morbidities and medications. Measures of calibration, discrimination, sensitivity, and specificity were calculated in the validation cohorts. There were 16,384 incident cases of oesophageal cancer in the derivation cohort (0.13% of 12.9 M). The predictors in the final algorithms were: age, BMI, Townsend deprivation score, smoking, alcohol, ethnicity, Barrett's oesophagus, hiatus hernia, H. pylori infection, use of proton pump inhibitors, anaemia, lung and blood cancer (with breast cancer in women). In the QResearch validation cohort in women the explained variation (R2) was 57.1%; Royston's D statistic 2.36 (95% CI 2.26-2.46); C statistic 0.859 (95% CI 0.849-0.868) and calibration was good. Results were similar in men. For the 20% at highest predicted risk, the sensitivity was 76%, specificity was 80.1% and the observed risk at 10 years was 0.76%. The results from the CPRD validation were similar. We have developed and validated a novel prediction algorithm to quantify the absolute risk of oesophageal cancer. The CanPredict algorithms could be used to identify high risk patients for targeted screening. Innovate UK and CRUK (grant 105857).


Modelling the effects of tides and storm surges on coastal aquifers using a coupled surface-subsurface approach.
Coastal aquifers are complex hydrologic systems because many physical processes interact: (i) variably saturated flow, (ii) spatial-temporal fluid density variations, (iii) tidal fluctuations, (iv) storm surges overtopping dykes, and (v) surface runoff of storm water. The HydroGeoSphere model is used to numerically simulate coastal flow dynamics, assuming a fully coupled surface-subsurface approach, accounting for all processes listed above. The diffusive wave approximation of the St. Venant equation is used to describe surface flow. Surface flow and salt transport are fully coupled with subsurficial variably saturated, variable-density flow and salt transport through mathematical terms that represent exchange of fluid mass and solute mass, respectively. Tides and storm surges induce a time-variant head that is applied to nodes of the surface domain. The approach is applied to real cases of tide and storm surge events. Tide simulation results confirm the existence of a recirculating zone, forming beneath the upper part of the intertidal zone. By monitoring the exchange fluid flux rates through the beach, it was found that the major inflow to the aquifer takes place at the upper part of the intertidal zone, which explains the formation of the recirculating zone. The recirculating zone is forming particularly during rising tide. Results from a storm surge simulation show that plume fingers develop below the flooded land surface. Natural remediation by seaward flowing freshwater is relatively slow, such that reducing the salt concentration in the aquifer down to drinking water standards takes up to 10 years.


Evolution and expression of BMP genes in flies.
Bone morphogenetic proteins (BMPs) play key roles in development. In Drosophila melanogaster, there are three BMP-encoding genes: decapentaplegic (dpp), glass bottom boat (gbb) and screw (scw). dpp and gbb are found in all groups of insects. In contrast, the origin of scw via duplication of an ancestral gbb homologue is more recent, with new evidence placing it within the Diptera. Recent studies show that scw appeared basal to the Schizophora, since scw orthologues exist in aschizan cyclorrhaphan flies. In order to further localise the origin of scw, we have utilised new genomic resources for the nematoceran moth midge Clogmia albipunctata (Psychodidae). We identified the BMP subclass members dpp and gbb from an early embryonic transcriptome and show that their expression patterns in the blastoderm differ considerably from those seen in cyclorrhaphan flies. Further searches of the genome of C. albipunctata were unable to identify a scw-like gbb duplicate, but confirm the presence of dpp and gbb. Our phylogenetic analysis shows these to be clear orthologues of dpp and gbb from other non-cyclorrhaphan insects, with C. albipunctata gbb branching ancestrally to the cyclorrhaphan gbb/scw split. Furthermore, our analysis suggests that scw is absent from all Nematocera, including the Bibionomorpha. We conclude that the gbb/scw duplication occurred between the separation of the lineage leading to Brachycera and the origin of cyclorrhaphan flies 200-150 Ma ago.


Interaction of meso-tetrakis(4-sulphonatophenyl)porphine with chitosan in aqueous solutions.
Interaction of meso-tetrakis(4-sulphonatophenyl)porphine (TPPS4) with chitosan (Mr approximately 400 kDa, N-acetyls approximately 20 mol.%) was studied in aqueous solutions. UV-vis absorption and circular dichroism (CD) spectroscopic titration of 10 micromol l-1 TPPS4 with chitosan demonstrated that an addition of the polysaccharide at appropriate concentrations and pH values induce and support self-aggregation of the macrocycles. The mode of aggregation was strongly dependent on pH: stacking (H-type) aggregates predominated at weak acidic conditions (pH 4.8-6.8) and tilted (J-type) aggregates at pH 2.5. At the intermediate pH value (3.6) both types of TPPS4 aggregates were detected. High amount of chitosan (>0.05 mmol l-1 of GlcN) disrupts H-aggregates forming monomeric porphyrin-chitosan complexes (pH 3.6-6.8), while J-aggregates (pH 2.5) are stable even at very high chitosan concentrations. CD titration experiments confirmed the formation of optically active species of TPPS4 in the presence of chitosan. The complex nature of CD bands assigned to both types of porphyrin aggregates indicated the occurrence of several chiral macrocyclic species dependently on pH value and chitosan concentration.


[The conjoined latissimus dorsi and parascapular free flap for reconstruction of extensive soft tissue defects].
Durable and resilient soft tissue reconstruction of vast defects of the extremities or the torso. Reconstruction of disproportionately large defects, particularly in cases of simultaneous bone and joint reconstruction. History of surgery or irradiation of upper back and axilla, impossibility of surgery under lateral positioning; relative contraindications in wheelchair users, hemiplegics, or amputees. General anesthesia and lateral positioning. First, the parascapular flap is harvested, with the initial skin incision made medially in order to identify the medial triangular space and the circumflex scapular artery. Flap raising then proceeds from caudal to cranial. Second, the latissimus dorsi is harvested, with the lateral border being dissected free first, before the thoracodorsal vessels are visualized on its undersurface. Flap raising then proceeds from caudal to cranial. Third, the parascapular flap is advanced through the medial triangular space. If the circumflex scapular and thoracodorsal vessels originate separately from the subscapular axis, an in-flap anastomosis is warranted. Subsequent microvascular anastomoses should be performed outside the zone of injury, typically in an end-to-end fashion of the vein and end-to-side fashion of the artery. Postoperative anticoagulation with low-molecular-weight heparin under anti-Xa monitoring (semitherapeutic in normal-risk and therapeutic in high-risk cases). Hourly clinical assessment of flap perfusion for 5 consecutive days, followed by stepwise relaxation of immobilization and commencement of dangling procedures in cases of lower extremity reconstruction. Between 2013 and 2018, 74 conjoined latissimus dorsi and parascapular flaps were transplanted to cover vast defects of the lower (n = 66) and upper extremity (n = 8). The mean defect size was 723 ± 482 cm2 and the mean flap size was 635 ± 203 cm2. Eight flaps required in-flap anastomoses for separate vascular origins. There was no case of total flap loss.


Reduction of infection rates in abdominal incisions by delayed wound closure techniques.
A low incidence of infection in abdominal wounds after contaminated, infected, and selected clean-contaminated operations was achieved after delayed wound closure of the skin and subcutaneous tissue. An effective method of delayed primary closure is described. Four days of open wound management with Xeroform gauze between the skin and subcutaneous tissue is followed on the 5th day be removal of the Xerform and skin approximation with Steri-Strips. Proper use of this technique is based upon appropriate assessment of wound contamination and infection risk factors. All contaminated and infected wounds are best managed with delayed primary closure and, when not possible, with healing by secondary intention. Delayed primary closure should be applied to clean-contaminated wounds if the patients are older than 60 years or have associated diabetes mellitus, malnutrition, or obesity.


Characterisation of CwpA, a putative glycosylphosphatidylinositol-anchored cell wall mannoprotein in the filamentous fungus Aspergillus niger.
Glycosylphosphatidylinositol (GPI)-anchored proteins in fungi are found at the cell surface, either as plasma membrane proteins (GPI-PMPs) or attached by a remnant of the GPI-anchor to the cell wall (GPI-CWPs). GPI-CWPs can be extracted from the cell wall by treatment with hydrofluoric acid (HF), which cleaves the phosphodiester bond that is present in the remnant of the GPI-anchor. The filamentous fungus Aspergillus niger contains at least seven HF-extractable cell wall mannoproteins. One gene encoding an HF-extractable cell wall mannoprotein, cwpA, was cloned and further characterised. The protein sequence of CwpA indicated the presence of two hydrophobic signal sequences both at the N-terminus and C-terminus of the protein, for entering the ER and the addition of a GPI-anchor, respectively. A CwpA-specific antiserum was raised and in combination with fractionation experiments, we show that this protein was abundantly present as an HF-extractable protein in the cell wall of A. niger.


Assessment of pharmacokinetics and pharmacodynamic effects related to abuse potential of a unique oral osmotic-controlled extended-release methylphenidate formulation in humans.
This was a double-blind, placebo-controlled, randomized, 5-period crossover study in 49 healthy subjects with a history of light (occasional) recreational stimulant use, to evaluate the abuse-related subjective effects of oral osmotic-controlled extended-release methylphenidate with comparable doses of immediate-release methylphenidate. Healthy subjects with a history of light recreational stimulant use were enrolled in the study if they demonstrated a positive response to a 20-mg dose of d-amphetamine and a negative placebo response. Enrolled subjects received single doses of placebo, 54 and 108 mg osmotic-controlled extended-release methylphenidate, and 50 and 90 mg immediate-release methylphenidate. For each treatment, pharmacokinetics, pharmacodynamics, and safety were assessed for 24 hours. Subjective data were collected through standard questionnaires and visual analog scales for positive, stimulant, negative, and other effects. Immediate-release and osmotic-controlled extended-release methylphenidate produced expected plasma concentration-time profiles of d-methylphenidate. Both doses of immediate-release methylphenidate (50 and 90 mg) produced statistically significantly higher subjective effects (eg, positive, stimulant) with respect to placebo for all measures. The higher osmotic-controlled extended-release methylphenidate dose of 108 mg also produced statistically significant differences from placebo for most measures. However, the most commonly prescribed therapeutic dose of osmotic-controlled extended-release methylphenidate (54 mg) did not produce significant differences from placebo for most measures. In addition, for comparable dose levels, osmotic-controlled extended-release methylphenidate produced lower positive and stimulant subjective effects than immediate-release methylphenidate, and low-dose immediate-release methylphenidate (50 mg) produced greater subjective effects than high-dose osmotic-controlled extended-release methylphenidate, with many effects demonstrating statistically significant differences. These data support the hypothesis that a formulation can modulate abuse potential by controlling the rate and extent of drug delivery.


Constitutive expression of the Trichoderma reesei beta-1,4-xylanase gene (xyn2) and the beta-1,4-endoglucanase gene (egl) in Aspergillus niger in molasses and defined glucose media.
The xylanase II (xyn2)- and endoglucanase I (egl)-encoding regions of Trichoderma reesei QM6a were successfully expressed in Aspergillus niger D15 under the transcriptional control of the glyceraldehyde-6-phosphate dehydrogenase (gpd) promoter from A. niger and the glaA terminator of Aspergillus awamori. A stable xyn2 transformant produced beta-xylanase activity of 8,000 nkat/ml and 5,000 nkat/ml in shake-flask cultures containing defined or 20% (v/v) molasses medium, respectively. The recombinant Xyn2 enzyme expressed highest activity at pH 5-6 and 50-60 degrees C and retained more than 75% of its activity after 3 h of incubation at 50 degrees C. A stable egl transformant produced endo-P-1,4-glucanase activity of 2,300 nkat/ml in shake-flask cultures containing defined media and about half the activity in 20% molasses medium. Maximum endoglucanase activity was obtained at pH 5 and 60 degrees C. Both Xyn2 and EgI retained >80% activity after incubation at 50 degrees C for 3 h. The heterologous Xyn2 and EgI represent a significant portion of the total extracellular proteins produced.


Physiological and molecular mechanisms of cobalt and copper interaction in causing phyto-toxicity to two barley genotypes differing in Co tolerance.
The combined effects of cobalt (Co) and copper (Cu) in their toxicity to plants is poorly studied although these two metals co-exist commonly in soil. In this study, a hydroponic experiment was carried out to investigate the effect of longer exposure of two barley genotypes differing in Co tolerance to the combined Co and Cu stress. The results confirmed the previous findings that Co accumulation in plant tissues was reduced by Cu presence, while Cu accumulation was stimulated by Co presence. Moreover, both single and combined treatments of Co and Cu reduced the mineral (Mn, Zn and K) uptake. Co and Cu applied alone or in combination at rate of 50 μM resulted in the significant reduction of plant growth and increase of oxidative stress (ROS and MDA), and meanwhile the capacity of scavenging active oxygen species (AOS) was enhanced, reflected by increased phytochelatin (PC) and glutathione (GSH and GSSG) content, as well as expression of the related genes (HvPCS1 and HvGR1). Yan66, a Co tolerant genotype was less affected in oxidative stress, and had higher AOS scavenging capacity in comparison with Ea52, a Co sensitive one. Among three HvSOD isoforms, only HvFeSOD expression was up-regulated in the combined treatment relative to control as well as the treatment of Co or Cu alone, while HvCuZnSOD and HvMnSOD were down-regulated and unaffected, respectively. In addition, the expressions of metal transporter genes (HvHMA2, HvHMA3 and HvHMA5) varied with genotype and metal treatments, with the extent being greater in Yan66 on the whole. The results suggest that upon longer exposure to the combined stress of Co and Cu, the greater phyto-toxicity than each element alone is associated with more Cu accumulation stimulated by Co and that, the higher regulation of transporter genes observed in Yan66 could in part explain for its higher metal tolerance ability.


Immunological identity of urinary alkaline phosphatase in health and disease.
Urinary alkaline phosphatase has been measured and an investigation has been undertaken involving starch gel electrophoresis after treatment of the enzyme with antisera raised against various human tissues. Urine was obtained from patients with bone and liver disease as well as pregnant women and normal persons. The enzyme level in urine in disease and pregnancy was raised above the normal but considerable overlap occurred. After electrophoresis the most usual finding was a single zone of activity although occasionally very minor zones were also present. Antiserum to kidney had the greatest affinity for the urinary enzyme. Decreasing affinity was found with intestinal and placental antiserum. No affinity was found with liver antiserum.


Postsurgical observations of mean hemispheric cerebral blood flow with patients receiving high-flow EC-IC bypass using a radial artery graft (preliminary report, one-year observation of 10 hemispheres).
When we encounter an inaccessible internal carotid artery lesion, we sacrifice the inaccessible internal carotid artery and, at the same time, establish a high-flow extracranial-to-intracranial arterial bypass (EC-IC bypass). Questions and concerns remain, however, about the dynamics of the patient's cerebral blood flow postoperatively. Ten hemispheres from 9 patients had received radial artery grafting as an EC-IC bypass with the simultaneous obstruction of the internal carotid artery. Their mean hemispheric cerebral blood flow (mCBF) was measured longitudinally during 1 year after surgery by single photon emission computed tomography (SPECT) and a 133Xenon-inhalation method. The serial mCBF values of the five hemispheres with giant aneurysm or paracavernous meningioma were attenuated during the postoperative 3-month period and then returned to the normal range. In only one case did the postoperative reduction of mCBF fail to return to normal; in addition there was a memory disturbance that had developed and progressed. Five hemispheres with the ICA stenosis did not present reduction in mCBF. In three out of five hemispheres, transient elevation of mCBF at 1 month after surgery was observed.


Surface Functionalization for Immobilization of Probes on Microarrays.
The microarray technology has been a tremendous advance in molecular-based testing methods for biochemical and biomedical applications. As a result, the immobilization techniques and grafting chemistries of biochemical molecules have experienced great progress. The particularities of the grafting techniques adapted to the microarray development will be presented here.


Efficacy of a standardized herbal formulation from Glycyrrhiza glabra L. as an adjuvant treatment in hospitalized patients with COVID-19: A Randomized Controlled trial.
As no specific pharmacological intervention has been known for COVID-19, medicinal plants may be a suitable candidate for management of this disease. The aim of this study was to investigate the efficacy of a herbal syrup from licorice as an adjuvant treatment in hospitalized patients with COVID-19. 213 hospitalized patients diagnosed with COVID-19 were assigned to receive either standardized licorice syrup as an adjuvant treatment plus standard care [Syrup Group (SYRUP), N = 91], or standard care alone [Standard Group (STANDARD), N = 104], for 7 days. The primary endpoint was duration of hospitalization in survivors. The secondary endpoints included 25% increase in oxygen saturation, C-reactive protein (CRP) difference and lymphocyte difference from baseline, number of death and number of patients transferred to ICU. Mean duration of admission was 5.24 days in SYRUP and 7.14 days in STANDARD (p < 0.001). Oxygen saturation increased in 86 of 91 patients (94.5%) in the licorice group, compared to 83 of 104 patients (79.8%) in the control group (p = 0.002). There was no significant difference between the two groups in the number of patients died during hospitalization (p = 0.837). Five patients in SYRUP and 16 patients in STANDARD were transferred to ICU (p < 0.026). Mean reduction in CRP (p < 0.001) and mean increase in the number of lymphocytes (p = 0.008) in SYRUP were significantly higher than STANDARD. Licorice syrup as an adjuvant treatment demonstrated promising results on duration of hospital admission, O2 saturation as well as inflammatory markers in COVID-19 patients; however, further clinical studies with larger sample size are suggested to achieve more conclusive results.


Determining the prevalence of human immunodeficiency virus-associated nephropathy (HIVAN) using proteinuria and ultrasound findings in a Nigerian paediatric HIV population.
HIV associated nephropathy (HIVAN) is the most common form of chronic kidney disease resulting directly from HIV infection. The true prevalence of HIVAN in the paediatric population of West Africa is unknown, largely due to lack of surveillance and reporting of kidney disease in HIV positive patients. This was a prospective study over a six month period( July to December 2008) conducted in the Infectious Disease Unit of the Department of Paediatrics, University of Uyo Teaching Hospital, Uyo, Nigeria involving all confirmed cases of paediatric HIV infection. Urine microalbuminuria using calculated urine albumin - creatinine ratio was determined and repeated in 4 weeks interval. CD4 count and renal ultrasonography was done for all the patients. The correlation of urine albumin - creatinine ratio with CD4 count, duration of treatment with highly active antiretroviral therapy (HAART) and association with clinical staging of the disease was also examined. Fifty - nine (60.2%) were males, thirty - nine (39.8%) were females with male to female ratio of 1.5:1. The prevalence rate of 31.6% HIVAN was found, out of which 3.1% had abnormal ultrasound findings. There was a significant correlation between CD4 count and urine albumin - creatinine ratio (r=-0.22, p=0.03). There was no correlation between urine albumin - creatinine ratio and duration on HAART (r=-0.10, p=0.31). Screening for microalbuminuria is essential for the early diagnosis and treatment of HIVAN in this age group.


The ostrich approach - Prognostic avoidance, strategies and barriers to assessing older hospital patients' risk of dying.
Predicting older patients' life expectancy is an important yet challenging task. Hospital aged care assessment teams advise treating teams on older patients' type and place of care, directly affecting quality of care. Yet, little is known about their experiences with prognostication. Twenty semi-structured interviews were conducted with seven geriatricians/ registrars, ten nurses and three allied health staff from aged care assessment teams across two hospitals in Melbourne, Australia. Data were analysed thematically. To generate prognoses, clinicians used analytical thinking, intuition, assessments from others, and pattern matching. Prognostic tools were an underutilised resource. Barriers to recognition of dying included: diffusion of responsibility regarding whose role it is to identify patients at end-of-life; lack of feedback about whether a prognosis was correct; system pressures to pursue active treatment and vacate beds; avoidance of end-of-life discussions; lack of confidence, knowledge and training in prognostication and pandemic-related challenges.


Computed tomography and magnetic resonance imaging of cerebral coenurosis.
A case of Coenurus cerebralis involving both cerebral hemispheres and the interpeduncular cistern is presented to illustrate the computed tomography (CT) and magnetic resonance imaging (MRI) features. In CT scans viable cysts appear as lucent lesions surrounded by a contrast-enhanced peripheral rim. By using multiple echo sequences the cyst content is characterized in magnetic resonance images by a cerebrospinal fluid-like intensity pattern.


Variability signatures distinguish verbal from nonverbal counting for both large and small numbers.
Humans appear to share with animals a nonverbal counting process. In a nonverbal counting condition, subjects pressed a key a numeral-specified number of times, while saying "the" at every press. The mean number of presses increased as a power function of the target number, with a constant coefficient of variation (c.v.), both within and beyond the proposed subitizing range (1-4 or 5), suggesting small numbers are represented on the same continuum as larger numbers and subject to the same noise process (scalar variability). By contrast, when subjects counted their presses out loud as fast as they could, the c.v. decreased as the inverse square root of the target value (binomial variability instead of scalar variability). The unexpected power-law relation between target value and mean number of presses in nonverbal counting suggests a new hypothesis about the development of the function relating number symbols to mental magnitudes.


Hsa_circ_0004287 inhibits macrophage-mediated inflammation in an N6-methyladenosine-dependent manner in atopic dermatitis and psoriasis.
Circular RNA (circRNA) has been implicated in various diseases; however, its role in atopic dermatitis (AD) or psoriasis remains unclear. We sought to determine the differential expression profiles of circRNAs in peripheral blood mononuclear cells between healthy controls and AD patients, and explore the mechanisms underlying the effects of circRNAs on the pathogenesis of AD. The differential expression profiles of circRNAs were analyzed by circRNA microarray. In vitro function and mechanisms by which circRNAs regulate macrophage-mediated inflammation were detected by reverse transcription quantitative PCR, Western blot analysis, RNA stability assay, immunoprecipitation, ELISA, and methylated RNA immunoprecipitation assay. In vivo roles of circRNAs were determined in 2,4-dinitrochlorobenzene (DNCB)-induced dermatitis and imiquimod (IMQ)-induced psoriasis mouse model. We identified a functional unknown circRNA hsa_circ_0004287 from 88750 circRNAs, which was upregulated in peripheral blood mononuclear cells of both AD and psoriasis patients, and was mainly expressed by macrophages under inflammatory conditions. Hsa_circ_0004287 inhibited M1 macrophage activation in vitro, and macrophage-specific overexpression of hsa_circ_0004287 alleviated skin inflammation in both AD- and psoriasis-like mice. Mechanistically, hsa_circ_0004287 reduced the stability of its host gene metastasis associated lung adenocarcinoma transcript 1 (MALAT1) by competitively binding to IGF2BP3 with MALAT1 in an N6-methyladenosine (m6A)-dependent manner. Lower levels of MALAT1 promoted the ubiquitination degradation of S100A8/S100A9, thereby impeding p38/mitogen-activated protein kinase phosphorylation and macrophage-mediated inflammation. hsa_circ_0004287 inhibits M1 macrophage activation in an m6A-dependent manner in AD and psoriasis, and may serve as a general therapeutic candidate for AD and psoriasis.


Probing the locomotor conundrum: descending the 'V' interneuron ladder.
The assembly of neuronal circuits involved in locomotor control in the mammalian spinal cord is influenced by genetic programs specifying four ventral (V) interneuron populations (V0-V3). In this issue of Neuron, Crone et al. and Zhang et al. make use of genetic tools to map connectivity patterns and to abolish the function of V2a and V3 interneurons. The absence of V2a interneurons reveals defects in left-right alternation during locomotion, whereas ablation of either V2a or V3 interneurons leads to disturbances in the precision and reliability of the motor output.


A review of tissue reference values used to assess the trace element status of farmed red deer (Cervus elaphus).
This paper reviews the principles for the establishment of biochemical reference criteria for assessing the trace element status of farmed livestock and summarises data for copper, selenium, vitamin B12 and iodine for farmed red deer. Enzootic ataxia and osteochondrosis occur when liver copper concentrations are below 60 micromol/kg fresh tissue, and serum copper concentrations are below 3-4 micromol/l. Growth responses to copper supplementation have been equivocal when blood copper concentrations were 3-4 micromol/l, but were significant when mean blood copper concentrations were 0.9-4.0 micromol/l. No antler growth or bodyweight response to copper supplementation was observed when blood ferroxidase levels averaged 10-23 IU/l (equivalent to serum copper concentrations of 6-13 micromol/l) and liver copper concentrations averaged 98 mumol/kg fresh tissue. These data suggest that 'deficient', 'marginal' and 'adequate' ranges for serum copper concentrations should be 5, 5-8, and 8 micromol/l, respectively, and those for liver copper concentrations should be 60, 60-100, and 100 micromol/kg, respectively. White muscle disease has been reported in young deer with blood and liver selenium concentrations of 84-140 nmol/l and 240-500 nmol/kg fresh tissue, respectively. No growth-rate response to selenium supplementation occurred in rising 1-year-old deer when blood selenium concentrations were less than 130 nmol/l, the range in which a growth-rate response would be expected in sheep. VITAMIN B12: Vitamin B12 concentrations in deer are frequently below 185 pmol/l without clinical or subclinical effects. No growth response was observed in young deer with vitamin B12 concentrations as low as 75-83 pmol/l. A growth response to cobalt/vitamin B12 supplementation occurs in lambs with serum vitamin B12 concentrations 336 pmol/l. Data that can be used to establish reference ranges for assessing trace element status in deer are limited. More robust reference values for farmed red deer need to be established through further studies relating biochemical data to health and performance.


[Association of epithermal growth factor receptor expression and its downstream gene mutation status with radiosensitivity of colorectal carcinoma cell lines in vitro].
To investigate the effect of epithermal growth factor receptor (EGFR) expression and K-ras, B-raf and PIK3CA mutation status on the radiosensitivity of human colorectal carcinoma (CRC) cell lines in vitro. Real-time RT-PCR was used to measure EGFR mRNA expression in nine human CRC cell lines, and K-ras, B-raf and PIK3CA mutation status of each CRC cell line was also identified respectively. After treatment with irradiation at graded dose, the cell viability was measured by clonogenic survival assay. The rate of cell apoptosis and cell cycle distribution were tested by flow cytometry. The cell morphology was observed with hoechst 33258 staining to analyze the correlation between EGFR mRNA expression and radiosensitivity of CRC cell lines. A positive correlation between EGFR mRNA expression and survival fraction of 2 Gy(SF2) was observed (r=0.717, P=0.030). Association was also identified between the mutation status of PIK3CA and radiosensitivity (t=2.401, P=0.047), while mutation status of K-ras and B-raf was not associated with radiosensitivity. At 48-hour after exposing to irradiation, the apoptosis rate of radiosensitive cell line (HCT116) was significantly increased in a dose-dependent manner (P<0.05), while the apoptosis rate of radioresistant cell line (HT29) was significantly increased only when radiation dose increased to 6 Gy. The ratio of G0/G1 phase was reduced significantly with the increase of radiation dose in radiosensitive cell line (HCT116, P<0.05), while this trend was not observed in radioresistant cell line (HT29, P>0.05). Over-expression of EGFR mRNA is correlated to radioresistance of human CRC cell lines, and mutation status of PIK3CA is closely related with radiosensitivity of CRC cells. The inhibition of apoptosis and G0/G1 arrest may induce the radioresistance of CRC cell lines.


How patients deal with an ambiguous medical test: Decision-making after genetic testing.
We know little about how patients make decisions when they receive a variant of uncertain significance result (VUS) from genetic testing. The purpose of this study was to elucidate a model of patient-informed decision-making after receiving a VUS result. Using an adapted Mental Models Approach, we conducted semi-structured interviews with women who received a VUS result from genetic testing for hereditary breast cancer (N = 20) to explore factors they believed were relevant to their decision-making. Two coders used a coding scheme informed by experts in hereditary breast cancer to conduct analysis. Inter-coder reliability was α = .86. Three overarching decision themes emerged from the interviews: managing ambiguity, medical risk management, and sharing results with others. While participants noted some difficulty understanding their result, genetic counselors' interpretations, psychosocial factors (e.g., risk perceptions), and competing extrinsic demands influenced their decisions. Complex influences affect patient decision-making after a VUS result from genetic testing and may encourage health protective behavior. Even patients who understand their test result could use support managing the ambiguity of their test result and sharing it with others.


Deep learning to predict elevated pulmonary artery pressure in patients with suspected pulmonary hypertension using standard chest X ray.
Accurate diagnosis of pulmonary hypertension (PH) is crucial to ensure that patients receive timely treatment. We hypothesized that application of artificial intelligence (AI) to the chest X-ray (CXR) could identify elevated pulmonary artery pressure (PAP) and stratify the risk of heart failure hospitalization with PH. We retrospectively enrolled a total of 900 consecutive patients with suspected PH. We trained a convolutional neural network to identify patients with elevated PAP (> 20 mmHg) as the actual value of PAP. The endpoints in this study were admission or occurrence of heart failure with elevated PAP. In an independent evaluation set for detection of elevated PAP, the area under curve (AUC) by the AI algorithm was significantly higher than the AUC by measurements of CXR images and human observers (0.71 vs. 0.60 and vs. 0.63, all p < 0.05). In patients with AI predicted PH had 2-times the risk of heart failure with PH compared with those without AI predicted PH. This preliminary work suggests that applying AI to the CXR in high risk groups has limited performance when used alone in identifying elevated PAP. We believe that this report can serve as an impetus for a future large study.


Long non-coding RNA Gm15441 attenuates hepatic inflammasome activation in response to PPARA agonism and fasting.
Exploring the molecular mechanisms that prevent inflammation during caloric restriction may yield promising therapeutic targets. During fasting, activation of the nuclear receptor peroxisome proliferator-activated receptor α (PPARα) promotes the utilization of lipids as an energy source. Herein, we show that ligand activation of PPARα directly upregulates the long non-coding RNA gene Gm15441 through PPARα binding sites within its promoter. Gm15441 expression suppresses its antisense transcript, encoding thioredoxin interacting protein (TXNIP). This, in turn, decreases TXNIP-stimulated NLR family pyrin domain containing 3 (NLRP3) inflammasome activation, caspase-1 (CASP1) cleavage, and proinflammatory interleukin 1β (IL1B) maturation. Gm15441-null mice were developed and shown to be more susceptible to NLRP3 inflammasome activation and to exhibit elevated CASP1 and IL1B cleavage in response to PPARα agonism and fasting. These findings provide evidence for a mechanism by which PPARα attenuates hepatic inflammasome activation in response to metabolic stress through induction of lncRNA Gm15441.


Evaluation of CHROMagar Candida for presumptive identification of clinically important Candida species.
CHROMagar Candida is a recently described and differential medium for the isolation and the presumptive identification of clinically important yeasts. We evaluated it with 262 yeast strains from clinical specimens, including 173 Candida albicans, 21 Candida tropicalis, 8 Candida krusei, 49 Candida glabrata, and 12 strains of other yeast species. Strains were presumptively identified on the basis of colony color and texture. These observations were compared with conventional identification results. Candida albicans was identified correctly in 170 (98%) of the 173 strains. A total of 46 of the 205 specimens that were plated on CHROMagar contained mixed cultures of yeast. Thirty-seven (80%) of these mixed cultures were not detected in the original specimens. CHROMagar Candida was useful for the rapid presumptive identification of Candida albicans and facilitated the recognition of mixed cultures. For other yeast species, it may provide additional information to laboratories that do not regularly perform identifications beyond the germ tube test.


Measuring Quality in Pediatric Endoscopy.
Measuring quality in endoscopy includes the assessment of appropriateness of a procedure and the skill with which it is performed. High-quality pediatric endoscopy is safe and efficient, used effectively to make proper diagnoses, is useful for excluding other diagnoses, minimizes adverse events, and is accompanied by appropriate documentation from beginning through end of the procedure. There are no standard quality metrics for pediatric endoscopy, but proposed candidates are both process and outcomes oriented. Both are likely to be used in the near future to increase transparency about patient outcomes, as well as to influence payments for the procedure.


Alemtuzumab-induced autoimmune thyroid events in patients with relapsing-remitting multiple sclerosis: A real-life and monocentric experience at a tertiary-level centre.
Alemtuzumab-induced autoimmune thyroid events (AIATEs) are the most common adverse effects observed in relapsing-remitting multiple sclerosis (RRMS) patients. This study aims to explore the clinical and biochemical characteristics of such AIATEs, and to examine the risk factors for their occurrence, particularly for the worst clinical phenotype of fluctuating Graves' disease (GD). We retrospectively analysed a real-life single-centre consecutive series of 57 RRMS patients treated with alemtuzumab whose clinical and biochemical parameters were collected before starting the treatment and then monthly during their follow-up. AIATEs developed in 39% of patients a mean 17 months ± 11 after the first cycle of alemtuzumab. The most common AIATEs were GD (64%), followed by Hashimoto's thyroiditis with hypothyroidism (23%), TSH-receptor-antibody (TRAb)-positive hypothyroidism (9%), and silent thyroiditis (4%). GD showed a fluctuating course in 57% of cases. Baseline positivity for anti-thyroperoxidase antibodies, and higher absolute titers of anti-thyroglobulin and anti-thyroperoxidase antibodies correlated significantly with the risk of developing AIATEs, but TRAb positivity did not. Higher TRAb titers at the time of GD being diagnosed correlated strongly with a greater risk of the fluctuating GD phenotype. On ROC curve analysis, we found that a cut-off of 7.3 IU/L could be used to predict the risk of developing a fluctuating GD, with a positive predictive value of 100%. TRAb levels measured with commercial automatic methods at the time of a patient being diagnosed with alemtuzumab-induced GD emerged as a novel biomarker for predicting a fluctuating disease phenotype, with an influence on subsequent therapeutic decisions and patients' follow-up.


Intraosseous lidocaine provides effective analgesia for percutaneous vertebroplasty of osteoporotic fractures.
To assess the safety and efficacy of intraosseous lidocaine (IL), in comparison with iv nalbuphine and propacetamol (NP) for analgesia during percutaneous vertebroplasty (PV) in order to avoid general anesthesia in elderly patients. Patients (age 68 +/- 13 yr, weight 66 +/- 6 kg) undergoing PV for osteoporotic fractures were randomized prospectively into two groups: NP (n=50) and IL (n=50). All patients were premedicated (oral hydroxyzine 1 mg.kg(-1)) and had skin infiltration with 5 mL of 1% lidocaine prior to vertebral puncture. Thirty minutes before the procedure, Group NP received, in a blinded manner, 50 mL of iv nalbuphine (0.3 mg.kg(-1)) and propacetamol (30 mg.kg(-1)) while Group IL received 50 mL of iv saline. During vertebral puncture, Groups NP and IL received, in a blinded manner, 1 mL.10 kg(-1) of intraosseous saline and 1% lidocaine respectively. Pain was assessed during vertebral puncture and cement injection with a four-point verbal rating scale. Additionally, lidocaine plasma kinetics were obtained in 11 IL patients. Analgesic efficacy was similar in the IL and NP groups (85 vs 84%). Group NP had more side effects. Lidocaine peak recorded concentration was 2.6 +/- 0.1 microg.mL(-1) i.e., about three times less than the reported toxic limits. IL is as effective as the association of iv NP for analgesia in PV. However, considering that both protocols were insufficient in about 15% of cases, other modalities are needed to further improve analgesia and avoid general anesthesia during vertebroplasty.


Are inflammatory markers associated with sarcopenia-related traits in older adults with sarcopenia? - A cross-sectional analysis of the ENHANce study.
To explore the relationship between inflammatory markers and sarcopenia-related traits in sarcopenic older adults. Baseline data of the ongoing Exercise and Nutrition for Healthy AgeiNg (ENHANce) study were used for a secondary, exploratory, cross-sectional analysis. ENHANce is a 5-armed triple blinded randomized controlled trial, in older adults (>65y) with sarcopenia defined according to the revised criteria of the European Working Group of Sarcopenia in Older People (EWGSOP2) aiming to assess the effect of combined anabolic interventions (protein supplement, omega-3 supplement and physical exercise) on physical performance, compared to single/placebo interventions. Inflammatory markers C-reactive protein (hs-CRP), albumin, interleukin-1β (IL-1β), IL-6, IL-8, and tumour necrosis factor-α (TNF-α) were assessed at baseline. Spearman's rho (ρ) correlation coefficients were calculated to associate these inflammatory markers with baseline sarcopenia-defining parameters (handgrip strength, chair stand test, appendicular lean mass [aLM], gait speed, Short Physical Performance Battery), physical activity (step count) and quality of life (SF-36, SarQoL). We included 40 sarcopenic subjects (15 men/25 women, age 77.1 ± 6.8 years). Contrary to expectations, the pro-inflammatory IL-1β correlated positively with handgrip strength (ρ: 0.376; p = 0.024) and IL-6 with aLM (ρ: 0.334; p = 0.0433). IL-6 inversely correlated with step count (ρ:-0.358; p = 0.048). Subgroup analysis revealed important gender differences. IL-8 inversely correlated with handgrip strength in women (ρ: -0.425; p = 0.034) but not in men. In contrast, pro-inflammatory cytokines CRP (ρ: -0.615; p = 0.019), IL-6 (ρ: -0.604; p = 0.029) and TNF-α (ρ: -0.615; p = 0.025) inversely correlated with the SF-36 physical component score in men but not in women. Although Inflammageing might play a role in sarcopenia-related traits, this exploratory study highlights an important role of gender. Future research should take this into account when elucidating the Inflammageing-sarcopenia interplay.


Revascularization and aneurysm surgery: current status.
Unclippable intracranial aneurysms are most effectively treated by hunterian ligation; however, the attendant risk of cerebral ischemia is significant. Many techniques have been used in an attempt to predict the safety of proximal vessel occlusion. Unfortunately, there is none that is risk-free and highly successful. A combination of stump pressure and cerebral blood flow measurements has been shown to be the most accurate in the acute assessment. In addition, recent studies have demonstrated that the long term risk of carotid ligation is significant. Extracranial-intracranial bypass grafting (EC-IC) has been shown to improve the safety of parent vessel ligation and is a low risk procedure. Whenever hunterian ligation is planned for the treatment of an intracranial aneurysm, EC-IC should be strongly considered.


Dostarlimab for recurrent mismatch repair-deficient endometrial cancer: A cost-effectiveness study.
Patients with recurrent endometrial cancer treated with carboplatin and paclitaxel whose disease progresses have few effective treatment options. Based on promising clinical trial data, the anti-programmed cell death 1 (anti-PD-1) antibody dostarlimab was recently granted accelerated approval for endometrial cancer by the US Food and Drug Administration. We developed a decision model to examine the cost-effectiveness of dostarlimab for patients with progressive/recurrent deficient mismatch repair (dMMR) endometrial cancer whose disease has progressed with first-line chemotherapy. Cost-effectiveness study. Hypothetical cohort of 6000 women with progressive/recurrent dMMR endometrial cancer. The initial decision point in the Markov model was treatment with dostarlimab, pembrolizumab or pegylated liposomal doxorubicin (PLD). Model probabilities, and cost and utility values were derived with assumptions drawn from published literature. Effectiveness was estimated as average quality-adjusted life years (QALYs) gained. One-way, two-way and probabilistic sensitivity analyses were performed to vary the assumptions across a range of plausible values. The primary outcome was the incremental cost-effectiveness ratio (ICER). Pegylated liposomal doxorubicin (PLD) was the least costly strategy, at $55,732, followed by dostarlimab ($151,533) and pembrolizumab ($154,597). Based on a willingness-to-pay threshold of $100,000/QALY, PLD was cost-effective compared with dostarlimab, with an ICER of $331,913 per QALY gained for dostarlimab, whereas pembrolizumab was ruled out by extended dominance (less effective, more costly), compared with dostarlimab. In one-way sensitivity analyses, dostarlimab was cost-effective when its cost was reduced to $4905 (52% reduction). These results were robust in a variety of sensitivity analyses. Dostarlimab is associated with greater survival compared with other treatments for women with recurrent dMMR endometrial cancer. Although the agent is substantially more costly, dostarlimab became cost-effective when its cost was reduced to $5489 per cycle.


Information-based analysis of the relationship between brain and facial muscle activities in response to static visual stimuli.
Human facial muscles react differently to different visual stimuli. It is known that the human brain controls and regulates the activity of the muscles. In this research, for the first time, we investigate how facial muscle reaction is related to the reaction of the human brain. Since both electromyography (EMG) and electroencephalography (EEG) signals, as the features of muscle and brain activities, contain information, we benefited from the information theory and computed the Shannon entropy of EMG and EEG signals when subjects were exposed to different static visual stimuli with different Shannon entropies (information content). Based on the obtained results, the variations of the information content of the EMG signal are related to the variations of the information content of the EEG signal and the visual stimuli. Statistical analysis also supported the results indicating that the visual stimuli with greater information content have a greater effect on the variation of the information content of both EEG and EMG signals. This investigation can be further continued to analyze the relationship between facial muscle and brain reactions in case of other types of stimuli.


Identification and genetic effect of haplotype in the bovine BMP7 gene.
Bone morphogenetic proteins (BMPs) are peptide growth factors belonging to the transforming growth factor-beta (TGF-β) superfamily, and some members of the BMP family support white adipocyte differentiation. In this study, we focused on the BMP7 which singularly promotes the differentiation of brown preadipocytes. Haplotypes involving 5 single nucleotide polymorphism (SNP) sites in the bovine BMP7 gene were identified and their effect on body weight was analyzed. 16 haplotypes and 18 combined haplotypes were revealed and the linkage disequilibrium was assessed in the cattle population with 602 individuals representing three main cattle breeds from China. The results showed that haplotypes 3, 10 and 14 were predominant and accounted for 75.64%, 69.85%, and 83.36% in Nanyang, Qinchuan and Jiaxian cattle breeds, respectively. The statistical analyses indicated that the SNP 1, 4, and 5 are associated with the body weight, body length, and heart girth at 12 and 24 months in Nanyang cattle population (P<0.05), whereas there is no significant association between their 16 haplotypes and 18 combined haplotypes. Our results provide evidence that some SNPs and haplotypes in BMP7 are associated with growth traits, and may be utilized as a genetic marker in marker-assisted selection for beef cattle breeding programs.


Serum Metabolomic Markers of Dairy Consumption: Results from the Atherosclerosis Risk in Communities Study and the Bogalusa Heart Study.
Dairy consumption is related to chronic disease risk; however, the measurement of dairy consumption has largely relied upon self-report. Untargeted metabolomics allows for the identification of objective markers of dietary intake. We aimed to identify associations between dietary dairy intake (total dairy, low-fat dairy, and high-fat dairy) and serum metabolites in 2 independent study populations of United States adults. Dietary intake was assessed with food frequency questionnaires. Multivariable linear regression models were used to estimate cross-sectional associations between dietary intake of dairy and 360 serum metabolites analyzed in 2 subgroups of the Atherosclerosis Risk in Communities study (ARIC; n = 3776). Results from the 2 subgroups were meta-analyzed using fixed effects meta-analysis. Significant meta-analyzed associations in the ARIC study were then tested in the Bogalusa Heart Study (BHS; n = 785). In the ARIC study and BHS, the mean age was 54 and 48 years, 61% and 29% were Black, and the mean dairy intake was 1.7 and 1.3 servings/day, respectively. Twenty-nine significant associations between dietary intake of dairy and serum metabolites were identified in the ARIC study (total dairy, n = 14; low-fat dairy, n = 10; high-fat dairy, n = 5). Three associations were also significant in BHS: myristate (14:0) was associated with high-fat dairy, and pantothenate was associated with total dairy and low-fat dairy, but 23 of the 27 associations significant in the ARIC study and tested in BHS were not associated with dairy in BHS. We identified metabolomic associations with dietary intake of dairy, including 3 associations found in 2 independent cohort studies. These results suggest that myristate (14:0) and pantothenate (vitamin B5) are candidate biomarkers of dairy consumption.


The DNA crosslink-induced S-phase checkpoint depends on ATR-CHK1 and ATR-NBS1-FANCD2 pathways.
The genetic syndrome Fanconi anemia (FA) is characterized by aplastic anemia, cancer predisposition and hypersensitivity to DNA interstrand crosslinks (ICLs). FA proteins (FANCs) are thought to work in pathway(s) essential for dealing with crosslinked DNA. FANCs interact with other proteins involved in both DNA repair and S-phase checkpoint such as BRCA1, ATM and the RAD50/MRE11/NBS1 (RMN) complex. We deciphered the previously undefined pathway(s) leading to the ICLs-induced S-phase checkpoint and the role of FANCs in this process. We found that ICLs activate a branched pathway downstream of the ATR kinase: one branch depending on CHK1 activity and the other on the FANCs-RMN complex. The transient slow-down of DNA synthesis was abolished in cells lacking ATR, whereas CHK1-siRNA-treated cells, NBS1 or FA cells showed partial S-phase arrest. CHK1 RNAi in NBS1 or FA cells abolished the S-phase checkpoint, suggesting that CHK1 and FANCs/NBS1 proteins work on parallel pathways. Furthermore, we found that ICLs trigger ATR-dependent FANCD2 phosphorylation and FANCD2/ATR colocalization. This study demonstrates a novel relationship between the FA pathway(s) and the ATR kinase.


Effects of repetitive passive movement on ankle joint on spinal reciprocal inhibition.
Repetitive passive movement (RPM) activates afferent Ia fibers. The input of afferent Ia fibers from antagonist muscle may modulate the extent of spinal reciprocal inhibition (RI). However, effects of RPM on RI remain unknown. We aimed to clarify these effects in 20 healthy adults. Four RPM tasks (40°/s, 80°/s, 120°/s, and 160°/s), with the range of ankle joint movement set to 40°, ranging from 10° in dorsiflexion to 30° in plantar flexion, were performed for 10 min. For measuring RI, a deep peroneal nerve as a conditioning stimulus, tibial nerve as a test stimulus, and three condition-test stimulus intervals (CTIs; single, 2 ms, and 20 ms) were used. The stimulation frequency was 0.3 Hz for 36 times (3 stimulation conditions × 12 sets). RI was measured before, immediately after, and 5, 10, 15, and 20 min (Pre, Post 5, 10, 15, and 20, respectively) after the task. The extent of reciprocal Ia inhibition (CTI 2 ms) significantly increased in Post 5 and 10 at RPM speed of ≥ 120°/s. The extent of D1 inhibition (CTI 20 ms) significantly increased in Post 5 and 10 at RPM speed of ≥ 80°/s, and continued to increase until Post 15 at RPM speed of 160°/s. The extent of RI was the highest at RPM speed of 160°/s for both Ia and D1. Therefore, high RPM may increase the extent of reciprocal Ia inhibition and D1 inhibition, suggesting that rapid movements affect RI by increasing the firing frequency from the muscle spindle to afferent Ia fibers.


Overexpression of mouse Mdm2 induces developmental phenotypes in Drosophila.
The Mdm2 proto-oncogene is amplified and over-expressed in a variety of tumors. One of the major functions of Mdm2 described to date is its ability to modulate the levels and activity of the tumor suppressor protein p53. Mdm2 binds to the N-terminus of p53 and, through its action as an E3 ubiquitin ligase, targets p53 for rapid proteasomal degradation. Mdm2 can also bind to other cellular proteins such as hNumb, E2F1, Rb and Akt; however, the biological significance of these interactions is less clear. To gain insight into the function of Mdm2 in vivo, we have generated a transgenic Drosophila strain bearing the mouse Mdm2 gene. Ectopic expression of Mdm2, using the UAS/GAL4 system, causes eye and wing phenotypes in the fly. Analysis of wing imaginal discs from third instar larvae showed that expression of Mdm2 induces apoptosis. Crosses did not reveal genetic interactions between Mdm2 and the Drosophila homolog of E2F, Numb and Akt. These transgenic flies may provide a unique experimental model for exploring the molecular interactions of Mdm2 in a developmental context.


Complete interruption of aortic arch diagnosed in adulthood: a case report.
Few patients with interrupted aortas survive into maturity, and the majority of instances are diagnosed in young children. There are only a few cases of this extremely rare total aortic interruption that survives into maturity, necessitating the substantial growth of collaterals to supply the descending aorta. Here, we describe a rare instance of an interrupted aorta in a 43-year male that presented in late adulthood with complete interruption of the aortic arch. The patient has remained symptom-free and without treatment. This case gives us an idea about how one can survive into adulthood with complete interruption of the aortic arch given that extensive collaterals are formed.


Eosinophilic colitis in a patient with acute myeloid leukemia after allogeneic bone marrow transplantation.
Eosinophilic colitis is a rare inflammatory disease characterized by eosinophilic infiltration of the colon and peripheral blood eosinophilia. We report on a case of eosinophilic colitis in a 29-year-old woman with acute myeloid leukemia following allogeneic bone marrow transplantation from her HLA-identical sister. To our knowledge, eosinophilic colitis has rarely been reported in association with allogeneic bone marrow transplantation.


Effects of auricular acupressure on depression and anxiety in older adult residents of long-term care institutions: A randomized clinical trial.
The current study was a single-blind clinical trial that evaluated the effect of auricular acupressure on older adults' depression and anxiety. Forty-seven older adults in long-term care institutions were randomly divided into two groups using block randomization. In the experimental group, patches with magnetic beads were pasted onto the auricular Shenmen acupoints for 14 days. In the control group, blank patches were used. Among the older adults in the study institutions, 82.09% scored ≥5 points on the Geriatric Depression Scale (GDS). Significant differences in GDS scores were observed between the two groups after both 7 and 14 days of treatment (p < 0.05). There was no significant difference (p < 0.05) in anxiety scores between the two groups at 7 days, but there was a significant difference after 14 days of the intervention (p < 0.05). Our results suggest that acupressure can reduce depression and anxiety among older adults in long-term care institutions.


Association of a CB1 cannabinoid receptor gene (CNR1) polymorphism with severe alcohol dependence.
Due to the involvement of the endogenous cannabinoid system in brain reward mechanisms a silent polymorphism (1359G/A; Thr453Thr) in the single coding exon of the CB1 human cannabinoid receptor gene (CNR1) was analysed in 121 severely affected Caucasian alcoholics and 136 most likely non-alcoholic controls. The observed frequency of the A allele was 31.2% for controls and 42.1% for alcoholics with severe withdrawal syndromes (P=0.010). Post-hoc exploration indicated that this allelic association resulted from an excess of the homozygous A/A genotype in patients with a history of alcohol delirium (P=0.031, DF 2), suggesting s an increased risk of delirium (OR=2.45, 95% CI 1.14--5.25). This finding suggests that the homozygous genotype CNR1 1359A/A confers vulnerability to alcohol withdrawal delirium.


FTIR spectroscopic, thermal and XRD characterization of hydroxyapatite from new natural sources.
The inorganic constituents of 5 different plants (leaves and stalks) were investigated by using Fourier transformer infrared spectroscopy (FTIR), X-ray diffraction (XRD) and thermal analysis including thermal gravimetric analysis (TGA), derivative thermogravimetry (DTG) and differential scanning calorimetry (DSC). These plants are Catha edulis (Khat), basil, mint, green tea and trifolium. The absorption bands of carbonate ions CO(3)(2-) was exhibited at 1446 cm(-1), and the phosphate ions PO(4)(3-) was assigned at 1105 and 1035 cm(-1). At high temperatures (600, 700 and 600°C) further absorption bands of the phosphate ions PO(4)(3-) was assigned at the frequencies 572, 617, 962, 1043 and 1110 cm(-1) and the vibrational absorption band of the carbonate ions CO(3)(2-) was assigned at 871, 1416 and 1461 cm(-1). X-ray diffraction and thermal analysis confirm the obtained results of FITR. Results showed that the main inorganic constituents of C. edulis and basil leaves are hydroxyapatite whereas the hydroxyapatite content in the other plant samples is less than that in case of C. edulis and basil plant leaves.


Spatial firing patterns of auditory neuron network modelling by computer simulation.
This communication examines, in digital computer simulated network, input signals and response patterns established at excitatory neurons' level i.e. the membrane potential of neuron soma. It is restricted to spatial patterns of the auditory neuron networks and time factor for nervous conduction and transmission is neglected compared with long maintained membrane potentials of neuron somas. The model analyzes the change in the spatial patterns of the membrane potential in the two dimensional networks of the auditory system. In order to evaluate the contribution of the various parameters, it is started that the simplest model has only one parameter, lateral inhibition. The other parameters are then added, one at a time, to successive models. The lateral inhibition is a necessary condition in the auditory nervous system if any sharpening of the response areas in the single neurons is to occur. A necessary condition for the validity of the model is that is should be applicable to the other senses such as vision and chemical patterns, taste. The threshold feature of auditory neurons aids in producing a sharpening in the neuron of the auditory relay nuclei. It does this clipping the spatial response patterns in one dimensional arrays of excitatory neurons. Recurrent inhibition seems a necessary condition in the sensory nervous system that any kinds of input signals are to be preserved over a wide range of stimulus intensity. In other words, this network has a wide dynamic range against any kinds of input signals. A simple self-recurrent negative feedback does not contribute to the sharpening, but more complex socalled averaged type does. A neuron network is capable of responding stably to stimuli with a wide range of intensity and with any kind of spatial patterns if there is a simple negative feedback mechanism. When there is no negative feedback, input signals soon disappear or saturate in the neuron network. Therefore, recurrent inhibition is the most important mechanism. Spontaneous activity appears to aid in the sharpening by providing a kind of contrast, that is by reducting the amount of activity in neurons adjacent to the excitatory area. Moreover, the effect of spontaneous activity in the model seems to make repples around the excitatory area and suggests that an introduction of activity at any stage of the networks, from whatever source for example reticulum formation and thalamus, might appreciably alter the response patterns at subsequent neuron network. This suggests that the mechanism of the consciousness that might be controlled by the thalamus and or reticular formation. These two dimensional neuron networks may be expanded to three dimensional neuron networks. The former might simulate the auditory nervous system while the latter might simulate the visual system.


Thiol modified bimodal mesoporous silica nanoparticles for removal and determination toxic vanadium from air and human biological samples in petrochemical workers.
Investigation of exposure to toxic vanadium (V) in petrochemical workers is very important for human health, and it must be removed and determined in workplace air and human biological samples. In this research, the enriched adsorbent based on the thiol modified bimodal mesoporous silica nanoparticle (HS-UVM7) was used for the extraction vanadium in human blood by the dispersive sonication ionic liquid micro solid phase extraction (DS-IL-μ-SPE) at pH of 4.5. In addition, the vanadium (V) was removed from the industrial workplace air based on HS-UVM7 adsorbent by the liquid-solid phase-gas removal (LSP-GR). In the static and dynamic system, the vanadium (V) was removed from artificial air with HS-UVM7 and compared with the polyvinyl chloride membrane (PCM, sorbent in 7300 NIOSH). The LSP-GR procedure based on HS-UVM7 had more recovery and adsorption capacity as compared to PCM. The adsorption capacity of HS-UVM7 and UVM7 adsorbents were obtained 144.1 mg g-1 and 23.3 mg g-1, respectively. In addition, the main parameters effected on extraction vanadium in blood samples and removal from air were studied and optimized by ET-AAS. The LOD, RSD%, linear range (LR) and enrichment factor (EF) was achieved 0.03 μg L-1, 3.1, 0.1-4.5 μg L-1 and 48.7, respectively for extraction of vanadium in 10 mL of blood samples by the DS-IL-MSPE procedure. The validation of the methodology was confirmed by standard addition to gas phase and using certified reference materials (CRM, NIST) or ICP-MS in human blood samples.


Chimeric Antigen Receptor (CAR) T Cell Therapy for Cancer. Challenges and Opportunities: An Overview.
The use of immunotherapy as an alternative treatment for cancer patients has become of great interest in the scientific community as it is required to overcome many of the currently unsolved problems such as tumor escape, immunosuppression and unwanted unspecific toxicity. The use of chimeric antigen receptor T cells has been a very successful strategy in some hematologic malignancies. However, the application of CAR T cells has been limited to solid tumors, and this has aimed the development of new generation of CARs with enhanced effectivity and specificity. Here, we review the state of the art of CAR T cell therapy with special emphasis on the current challenges and opportunities.


MR angiography in the follow-up of coiled cerebral aneurysms after treatment with Guglielmi detachable coils.
Intra-arterial digital subtraction angiography (DSA) has been considered the gold standard examination in the follow-up of patients treated with Guglielmi detachable coils (GDCs). However, DSA is an invasive and expensive investigation and results in exposure to ionising radiation to both patient and operator. The aim of this study was to compare MR angiography (MRA) with DSA with regard to patency of the occlusion of aneurysms following GDC treatment. We performed 75 MRA and DSA examinations on 51 patients treated with GDCs. The examinations were performed 3-36 months after embolisation and the interval between MRA and DSA was less than 1 week. Hard copies of both studies were interpreted retrospectively and independently for residual flow within the aneurysm, residual aneurysmal neck, and parent and branch vessel flow. Patency status of parent and branch vessel flow was correctly identified with MRA in all patients except 1. The sensitivity of MRA in revealing residual flow within the aneurysm was 97%. The specificity in ruling out residual flow within the aneurysm was 91%. MRA may replace DSA in the long-term follow-up of coiled cerebral aneurysms. The initial follow-up examination should, however, include both modalities.


Spectral changes of the fluorescent acriflavine--(Feulgen) DNA dye complex at different temperatures: application of a new cryostat for measurements in reflected and transmitted light.
The fluorometric behaviour of cellular objects is influenced during excitation by two nearly independent phenomena: (1) by the photochemical reaction of the DNA/AF dye complex, and (2) by the energy transfer among several DNA/AF dye complexes. Both processes show a distinct temperature-dependent behaviour and can therefore be characterized by the analysis of the fluorescence spectra at different temperatures. All microfluorometric measurements were performed with a self-constructed cooling device. The cryostat permits measurements of the cellular fluorescence within a range of temperatures between 4 K and 300 K. The cooling unit operates in accordance with the 'Continuous Flow Principle' and allows the application of objectives up to a numerical aperture of 0.6.


Th2 and regulatory immune reactions contribute to IgG4 production and the initiation of Mikulicz disease.
Mikulicz disease has been considered to be a subtype of Sjögren's syndrome (SS). However, recent studies have suggested that Mikulicz disease is an IgG4-related disease and is distinguishable from SS. In addition, it has been reported that both interleukin-4 (IL-4) and IL-10 induce IgG4 production and inhibit IgE. This study was undertaken to examine the expression of these cytokines in patients with Mikulicz disease and patients with SS. Labial salivary gland (LSG) sections from 15 patients with Mikulicz disease and 18 patients with SS were examined for subsets of the infiltrating lymphocytes, expression patterns of messenger RNA (mRNA) for cytokines/chemokines, and relationships between the IgG4:IgG ratio and the expression of mRNA for IL-4 or IL-10. Immunohistochemical analysis showed lymphocyte infiltration of various subsets in the LSGs of SS patients, and the selective infiltration of IgG4-positive plasma cells and Treg cells in the LSGs of Mikulicz disease patients. The levels of mRNA for both Th1 and Th2 cytokines and chemokines in LSGs from patients with SS were significantly higher than in controls, while the expression of both Th2 and Treg cells was significantly higher in the patients with Mikulicz disease than in controls. Furthermore, the expression of IL-4 or IL-10 in the LSGs was correlated with the IgG4:IgG ratio. These results suggest that the pathogenesis of Mikulicz disease is different from that of SS. Mikulicz disease is a unique inflammatory disorder characterized by Th2 and regulatory immune reactions that might play key roles in IgG4 production.


Biocompatible in-tube solid-phase microextraction coupled to HPLC for the determination of angiotensin II receptor antagonists in human plasma and urine.
A poly (methacrylic acid-ethylene glycol dimethacrylate, MAA-EGDMA) monolithic capillary was used for the in-tube solid-phase microextraction (in-tube SPME) of several angiotensin II receptor antagonists (ARA-IIs) from human plasma and urine. Under the optimized extraction condition, the protein component of the biological sample was flushed through the monolithic capillary, while the analytes were successfully trapped. Coupled to HPLC with fluorescence detection, this on-line in-tube SPME method was successfully applied for the determination of candesartan, losartan, irbesartan, valsartan, telmisartan, and their detection limits were found to be 0.1-15.3ng/mL and 0.1-15.2ng/mL in human plasma and urine, respectively. The method was linear over the range of 0.5-200ng/mL for telmisartan, 5-2000ng/mL for candesartan and irbesartan, 10-2000ng/mL for valsartan, and 50-5000ng/mL for losartan with correlation coefficients being above 0.9985 in plasma sample and above 0.9994 in urine sample. The method reproducibility was evaluated at three concentration levels, resulting in the R.S.D. <7%. The poly (MAA-EGDMA) monolithic capillary was demonstrated to be robust and biocompatible by using direct injections of biological samples.


Inhibition of phosphodiesterase-4 in the spinal dorsal horn ameliorates neuropathic pain via cAMP-cytokine-Cx43 signaling in mice.
The spinal phosphodiesterase-4 (PDE4) plays an important role in chronic pain. Inhibition of PDE4, an enzyme catalyzing the hydrolysis of cyclic adenosine monophosphate AMP (cAMP), produces potent antinociceptive activity. However, the antinociceptive mechanism remains largely unknown. Connexin43 (Cx43), a gap junction protein, has been shown to be involved in controlling pain transduction at the spinal level; restoration of Cx43 expression in spinal astrocytes to the normal levels reduces nerve injury-induced pain. Here, we evaluate the novel mechanisms involving spinal cAMP-Cx43 signaling by which PDE4 inhibitors produce antinociceptive activity. First, we determined the effect of PDE4 inhibitors rolipram and roflumilast on partial sciatic nerve ligation (PSNL)-induced mechanical hypersensitivity. Next, we observed the role of cAMP-Cx43 signaling in the effect of PDE4 inhibitors on PSNL-induced mechanical hypersensitivity. Single or repeated, intraperitoneal or intrathecal administration of rolipram or roflumilast significantly reduced mechanical hypersensitivity in mice following PSNL. In addition, repeated intrathecal treatment with either of PDE4 inhibitors reduced PSNL-induced downregulation of cAMP and Cx43, and upregulation of proinflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β. Furthermore, the antinociceptive effects of PDE4 inhibitors were attenuated by the protein kinase A (PKA) inhibitor H89, TNF-α, or Cx43 antagonist carbenoxolone. Finally, PSNL-induced upregulation of PDE4B and PDE4D, especially the PDE4B subtype, was reduced by treatment with either of the PDE4 inhibitors. The results suggest that the antinociceptive effect of PDE4 inhibitors is contributed by increasing Cx43 expression via cAMP-PKA-cytokine signaling in the spinal dorsal horn.