Optimization of ultrasonic-assisted extraction of cordycepin from Cordyceps militaris using orthogonal experimental design.
This study reports on the optimization of the extraction conditions of cordycepin from Cordyceps militaris by using ultrasonication. For this purpose, the orthogonal experimental design was used to investigate the effects of factors on the ultrasonic-assisted extraction (UAE). Four factors: extraction time (min), ethanol concentration (%), extraction temperature (°C) and extraction frequency (kHz), were studied. The results showed that the highest cordycepin yield of 7.04 mg/g (86.98% ± 0.23%) was obtained with an extraction time of 60 min, ethanol concentration of 50%, extraction temperature of 65 °C and extraction frequency of 56 kHz. It was found that the cordycepin extraction yield increased with the effect of ultrasonication during the extraction process. Therefore, UAE can be used as an alternative to conventional immersion extraction with respect to the recovery of cordycepin from C. militaris, with the advantages of shorter extraction time and reduced solvent consumption.


High sensitivity analysis of amyloid-beta peptide composition in amyloid deposits from human and PS2APP mouse brain.
Cortical amyloid-beta (Abeta) deposition is considered essential in Alzheimer's disease (AD) and is also detectable in nondemented individuals with pathologic aging (PA). The present work presents a detailed analysis of the Abeta composition in various plaque types from human AD and PA cases, compared with plaque Abeta isolated from PS2APP mice. To determine minute amounts of Abeta from 30 to 50 laser-dissected amyloid deposits, we used a highly sensitive mass spectrometry procedure after restriction protease lysyl endopeptidase (Lys-C) digestion. This approach allowed the analysis of the amino-terminus and, including a novel ionization modifier, for the first time the carboxy-terminus of Abeta at a detection limit of approximately 200 fmol. In addition, full length Abeta 40/42 and pyroglutamate 3-42 were analyzed using a highly sensitive urea-based Western blot procedure. Generally, Abeta fragments were less accessible in human deposits, indicative of more posttranslational modifications. Thioflavine S positive cored plaques in AD were found to contain predominantly Abeta 42, whereas thioflavine S positive compact plaques and vascular amyloid consist mostly of Abeta 40. Diffuse plaques from AD and PA, as well as from PS2APP mice are composed predominantly of Abeta 1-42. Despite biochemical similarities in human and PS2APP mice, immuno-electron microscopy revealed an extensive extracellular matrix associated with Abeta fibrils in AD, specifically in diffuse plaques. Amino-terminal truncations of Abeta, especially pyroglutamate 3-40/42, are more frequently found in human plaques. In cored plaques we measured an increase of N-terminal truncations of approximately 20% between Braak stages IV to VI. In contrast, diffuse plaques of AD and PA cases, show consistently only low levels of amino-terminal truncations. Our data support the concept that diffuse plaques represent initial Abeta deposits but indicate a structural difference for Abeta depositions in human AD compared with PS2APP mice already at the stage of diffuse plaque formation.


Non-Hodgkin's lymphomata: clinical features in relation to histology.
An analysis is given of the clinico-pathological correlations of non-Hodgkin's lymphomata in 332 patients referred to the Netherlands Cancer Institute in Amsterdam and the Rotterdam Radiotherapy Institute. Clinical staging proved to be an important prognostic index. In Stage I the 5 year survival was 55%, in Stage II 25% and in Stages III and IV less than 10%. The presence of follicular structures in non-Hodgkin's lymphomata has similarly an important prognostic significance especially in Stage I and II. In lymphocytic lymphomata a larger cell size is correlated with less favourable prognosis. The presence of macrophages in non-Hodgkin's lymphomata is found in patients with short survival. The histiocytic lymphomata have a different survival pattern from the lymphocytic lymphomata in Stages I and II.


Production of HO2 and OH radicals from near-UV irradiated airborne TiO2 nanoparticles.
The production of gas-phase hydroperoxyl radicals, HO2, is observed directly from sub-micron airborne TiO2 nanoparticles irradiated by 300-400 nm radiation. The rate of HO2 production as a function of O2 pressure follows Langmuir isotherm behaviour suggesting O2 is involved in the production of HO2 following its adsorption onto the surface of the TiO2 aerosol. Reduction of adsorbed O2 by photogenerated electrons is likely to be the initial step followed by reaction with a proton produced via oxidation of adsorbed water with a photogenerated hole. The rate of HO2 production decreased significantly over the range of relative humidities between 8.7 and 36.9%, suggesting competitive adsorption of water vapour inhibits HO2 production. From the data, the adsorption equilibrium constants were calculated to be: KO2 = 0.27 ± 0.02 Pa-1 and KH2O = 2.16 ± 0.12 Pa-1 for RH = 8.7%, decreasing to KO2 = 0.18 ± 0.01 Pa-1 and KH2O = 1.33 ± 0.04 Pa-1 at RH = 22.1%. The increased coverage of H2O onto the TiO2 aerosol surface may inhibit HO2 production by decreasing the effective surface area of the TiO2 particle and lowering the binding energy of O2 on the aerosol surface, hence shortening its desorption lifetime. The maximum yield (i.e. when [O2] is projected to atmospherically relevant levels) for production of gas-phase HO2, normalised for surface area and light intensity, was found to be at a RH of 8.7% for the 80% anatase and 20% rutile formulation of TiO2 used here. This yield decreased to as the RH was increased to 22.1%. Using this value, the rate of production of HO2 from TiO2 surfaces under atmospheric conditions was estimated to be in the range 5 × 104-1 × 106 molecule cm-3 s-1 using observed surface areas of mineral dust at Cape Verde, and assuming a TiO2 fraction of 4.5%. For the largest loadings of dust in the troposphere, the rate of this novel heterogeneous production mechanism begins to approach that of HO2 production from the gas-phase reaction of OH with CO in unpolluted regions. The production of gas-phase OH radicals could only be observed conclusively at high aerosol surface areas, and was attributed to the decomposition of H2O2 at the surface by photogenerated electrons.


A hybrid toxin from bacteriophage f1 attachment protein and colicin E3 has altered cell receptor specificity.
A hybrid protein was constructed in vitro which consists of the first 372 amino acids of the attachment (gene III) protein of filamentous bacteriophage f1 fused, in frame, to the carboxy-terminal catalytic domain of colicin E3. The hybrid toxin killed cells that had the F-pilus receptor for phage f1 but not F- cells. The activity of the hybrid protein was not dependent upon the presence of the colicin E3 receptor, BtuB protein. The killing activity was colicin E3 specific, since F+ cells expressing the colicin E3 immunity gene were not killed. Entry of the hybrid toxin was also shown to depend on the products of tolA, tolQ, and tolR which are required both for phage f1 infection and for entry of E colicins. TolB protein, which is required for killing by colicin E3, but not for infection by phage f1, was also found to be necessary for the killing activity of the hybrid toxin. The gene III protein-colicin E3 hybrid was released from producing cells into the culture medium, although the colicin E3 lysis protein was not present in those cells. The secretion was shown to depend on the 18-amino-acid-long gene III protein signal sequence. Deletion of amino acids 3 to 18 of the gene III moiety of the hybrid protein resulted in active toxin, which remained inside producing cells unless it was mechanically released.


Cellulosimicrobium protaetiae sp. nov., isolated from the gut of the larva of Protaetia brevitarsis seulensis.
A Gram-stain-positive, non-spore-forming, yellow-pigmented, non-motile, non-flagellated, facultative anaerobic and rod-shaped bacterial strain, designated BI34T, was isolated from the gut of the larva of Protaetia brevitarsis seulensis. Strain BI34T grew at 15-40 °C (optimum, 37 °C), at pH 6.5-9.0 (optimum, pH 7.5) and in the presence of 0-7 % (w/v) NaCl (optimum, 2 %). Based on the results of 16S rRNA gene sequence analysis, strain BI34T belonged to the phylum Actinobacteria and was closely related to Cellulosimicrobium funkei NBRC 104118T (99.3 %), Cellulosimicrobium cellulans NBRC 15516T (99.1 %), Cellulosimicrobium composti BIT-GX5T (99.0 %), Cellulosimicrobium fucosivorans SE3T (99.0 %), Cellulosimicrobium marinum NBRC 110994T (98.4 %) and Cellulosimicrobium terreum DS-61T (97.0 %). The genome to genome relatedness of the average nucleotide identity (ANI) and the digital DNA-DNA hybridization (dDDH) values calculated by the Genome-to-Genome Distance Calculator between strain BI34T and its related species mentioned above were lower than the threshold of 95 and 70 % for speciation, respectively. The predominant menaquinone of strain BI34T contained MK-9(H4), and the major fatty acids were anteiso-C15 : 0, C16 : 0 and anteiso-C17 : 0. Strain BI34T had diphosphatidylglycerol and phosphatidylglycerol as major polar lipids. The whole-cell sugars were galactose, glucose and ribose, and the cell-wall peptidoglycan contained lysine, alanine, aspartic acid and glutamic acid. The DNA G+C content of strain BI34T was 73.8 mol%. The difference in physiological and biochemical characteristics and the below-threshold values of genome-to-genome relatedness indicate that strain BI34T represents a novel species in the genus Cellulosimicrobium, for which the name Cellulosimicrobium protaetiae sp. nov. is proposed. The type strain is BI34T (=KCTC 49302T=NBRC 114073T).


The Global Regulators Lrp, LeuO, and HexA Control Secondary Metabolism in Entomopathogenic Bacteria.
Photorhabdus luminescens TTO1 and Xenorhabdus nematophila HGB081 are insect pathogenic bacteria and producers of various structurally diverse bioactive natural products. In these entomopathogenic bacteria we investigated the role of the global regulators Lrp, LeuO, and HexA in the production of natural products. Lrp is a general activator of natural product biosynthesis in X. nematophila and for most compounds in TTO1. Microarray analysis confirmed these results in X. nematophila and enabled the identification of additional biosynthesis gene clusters (BGC) regulated by Lrp. Moreover, when promoters of two X. nematophila BGC were analyzed, transcriptional activation by Lrp was observed. In contrast, LeuO in X. nematophila and P. luminescens has both repressing and activating features, depending on the natural product examined. Furthermore, heterologous overexpression of leuO from X. nematophila in the closely related Xenorhabdus szentirmaii resulted in overproduction of several natural products including novel compounds. The presented findings could be of importance for establishing a tool for overproduction of secondary metabolites and subsequent identification of novel compounds.


Magnetic resonance spectra of hyperpolarized (129)xe in human blood and living rat chest.
We constructed a gas polarization system to test the feasibility of using hyperpolarized (129)Xe gas as an NMR (nuclear magnetic resonance) probe to explore brain function. Both in vitro and in vivo experiments were performed with a 4.7 T NMR spectrometer. Xenon spectra from human blood confirmed the existence of two peaks corresponding to red blood cells and plasma. In rat studies, three peaks at around 200 ppm were observed. Our results are consistent with previously reported data.


SO2-Driven In Situ Formation of Superstable Hg3Se2Cl2 for Effective Flue Gas Mercury Removal.
Flue gas mercury removal is mandatory for decreasing global mercury background concentration and ecosystem protection, but it severely suffers from the instability of traditional demercury products (e.g., HgCl2, HgO, HgS, and HgSe). Herein, we demonstrate a superstable Hg3Se2Cl2 compound, which offers a promising next-generation flue gas mercury removal strategy. Theoretical calculations revealed a superstable Hg bonding structure in Hg3Se2Cl2, with the highest mercury dissociation energy (4.71 eV) among all known mercury compounds. Experiments demonstrate its unprecedentedly high thermal stability (>400 °C) and strong acid resistance (5% H2SO4). The Hg3Se2Cl2 compound could be produced via the reduction of SeO32- to nascent active Se0 by the flue gas component SO2 and the subsequent combination of Se0 with Hg0 and Cl- ions or HgCl2. During a laboratory-simulated experiment, this Hg3Se2Cl2-based strategy achieves >96% removal efficiencies of both Hg0 and HgCl2 enabling nearly zero Hg0 re-emission. As expected, real mercury removal efficiency under Se-rich industrial flue gas conditions is much more efficient than Se-poor counterparts, confirming the feasibility of this Hg3Se2Cl2-based strategy for practical applications. This study sheds light on the importance of stable demercury products in flue gas mercury treatment and also provides a highly efficient and safe flue gas demercury strategy.


Spatio-temporal patterning of arginyl-tRNA protein transferase (ATE) contributes to gametophytic development in a moss.
The importance of the arginyl-tRNA protein transferase (ATE), the enzyme mediating post-translation arginylation of proteins in the N-end rule degradation (NERD) pathway of protein stability, was analysed in Physcomitrella patens and compared to its known functions in other eukaryotes. We characterize ATE:GUS reporter lines as well as ATE mutants in P. patens to study the impact and function of arginylation on moss development and physiology. ATE protein abundance is spatially and temporally regulated in P. patens by hormones and light and is highly abundant in meristematic cells. Further, the amount of ATE transcript is regulated during abscisic acid signalling and downstream of auxin signalling. Loss-of-function mutants exhibit defects at various levels, most severely in developing gametophores, in chloroplast starch accumulation and senescence. Thus, arginylation is necessary for moss gametophyte development, in contrast to the situation in flowering plants. Our analysis further substantiates the conservation of the N-end rule pathway components in land plants and highlights lineage-specific features. We introduce moss as a model system to characterize the role of the NERD pathway as an additional layer of complexity in eukaryotic development.


An unusual cause of hearing loss in an 81-year-old man.
An 81-year-old man presented to accident and emergency with a recent history of rapidly progressive bilateral hearing loss. Further questioning revealed recent sinusitis, fevers and general malaise. Initial bloods showed raised inflammatory markers, raised creatinine, low albumin and urine dip was positive for blood and protein. Chest x-ray showed faint bilateral mid-zone infiltrates. He was treated with intravenous methylprednisolone (500 mg×3) for presumed Wegener's granulomatosis which was later confirmed by c-anti-neutrophil cytoplasmic antibodies testing (proteinase-3 positive) and supportive findings on CT chest and sinuses. He was discharged after 10 days in hospital on oral prednisolone and cyclophosphamide. He is continuing treatment and seen regularly in outpatient follow-up. Unfortunately his hearing has not fully recovered and he is awaiting a hearing aid.


Combined strategies on the treatment of cerebellar arteriovenous malformation.
Cerebellar arteriovenous malformations (AVMs) comprise 10%-15% of all intracranial AVMs and have a higher risk for morbidity and mortality than supratentorial AVMs. Patients with cerebellar AVMs present with hemorrhage more often than patients with cerebral AVMs, justifying an interventional treatment. Patient outcome can be predicted with specific grade systems, guiding vascular neurosurgeons in decision-making. The authors present the case of a 42-year-old man incidentally diagnosed with an unruptured cerebellar inferior vermian AVM, which was managed through a combined strategy of preoperative endovascular embolization of the main arterial feeders followed by microsurgical resection via midline suboccipital craniotomy, with a favorable outcome. The video can be found here: https://youtu.be/3WESejZbk90.


Attitude of outpatients with neuromuscular diseases in Japan to pain and use of analgesics.
The prevalence of pain and its impact on outpatients with neuromuscular disease, and their attitude towards the use of analgesics were studied. Seventy-eight outpatients at the university hospital, Tokyo, diagnosed with Parkinson's disease, spinocerebellar degeneration, amyotrophic lateral sclerosis, or multiple sclerosis were asked whether they had experienced pain in the preceding week. The Brief Pain Inventory, Japanese version was used to interview participants reporting pain, about its intensity and interference with activities, the way they dealt with it, attitudes to pain and use of analgesics, and desire for treatment. Forty-six participants experienced pain in the preceding week (59%). The mean pain intensity was 4.1 out of 10, and 20% of participants reported that the degree of interference with mobility was at least 6 out of 10. Most participants dealt with their pain without medication, by changing posture frequently or massage. Approximately 80% of participants regarded pain as something they should endure. Half of the participants wanted more information on methods for pain relief. Approximately 80% of participants were anxious about adverse reactions of analgesics. These findings suggest that medical staffs should provide appropriate information and educate their patients.


Predictors of occult metastasis in clinical stage I nonseminoma: a systematic review.
Patients with clinical stage I nonseminomatous testicular germ cell tumor should ideally receive adjuvant therapy only when they are at high risk for occult metastasis. We aimed to quantify the importance of predictors for occult metastasis by performing a systematic review of the relevant literature. In addition, we reviewed published multivariable models and risk-adapted treatment policies. We identified 23 publications between 1979 and 2001, reporting a total of 2,587 patients. Twenty-nine percent of the patients (759 of 2,587 patients) had occult metastases, which was diagnosed either at retroperitoneal lymph node dissection (n = 193) or during follow-up (n = 566). Odds ratios (OR) were pooled using meta-analysis techniques. The presence of vascular invasion of the primary tumor cells had the strongest effect (OR, 5.2; 95% CI, 4.0 to 6.8). Immunohistochemical staining of the primary tumor cells with the MIB-1 monoclonal antibody showing proliferative activity was a promising predictor (OR, 4.7; 95% CI, 2.0 to 11). Intermediate effects were found for embryonal carcinoma in the primary tumor (OR, 2.9; 95% CI, 2.0 to 4.4) and a high pathologic stage of the tumor (OR, 2.6; 95% CI, 1.8 to 3.8). Size of the primary tumor and age of the patient had weaker though also statistically significant associations with occult metastasis. Until now, multivariable models often included vascular invasion and embryonal carcinoma with one or two weaker predictors. None of the published risk-adapted treatment policies included MIB-1 staining. Several strong predictors for occult metastasis were identified. A risk-adapted treatment policy should be developed that incorporates all relevant predictors so that adjuvant therapy is targeted better to those with occult metastases.


Suitability of three indicators measuring the quality of coordination within hospitals.
Coordination within hospitals is a major attribute of medical care and influences quality of care. This study tested the validity of 3 indicators covering two key aspects of coordination: the transfer of written information between professionals (medical record content, radiology exam order) and the holding of multidisciplinary team meetings during treatment planning. The study was supervised by the French health authorities (COMPAQH project). Data for the three indicators were collected in a panel of 30 to 60 volunteer hospitals by 6 Clinical Research Assistants. The metrological qualities of the indicators were assessed: (i) Feasibility was assessed using a grid of 19 potential problems, (ii) Inter-observer reliability was given by the kappa coefficient () and internal consistency by Cronbach's alpha test, (iii) Discriminatory power was given by an analysis of inter-hospital variability using the Gini coefficient as a measure of dispersion. Overall, 19281 data items were collected and analyzed. All three indicators presented acceptable feasibility and reliability (, 0.59 to 0.97) and showed wide differences among hospitals (Gini, 0.08 to 0.11), indicating that they are suitable for making comparisons among hospitals. This set of 3 indicators provides a proxy measurement of coordination. Further research on the indicators is needed to find out how they can generate a learning process. The medical record indicator has been included in the French national accreditation procedure for healthcare organisations. The two other indicators are currently being assessed for inclusion.


New formulations of demineralized bone matrix as a more effective graft alternative in experimental posterolateral lumbar spine arthrodesis.
A rabbit model of posterolateral intertransverse process spine arthrodesis was used. To determine the efficacy of two new formulations of demineralized bone matrix. The flowable gel form of Grafton (Osteotech, Eatontown, NJ) demineralized bone matrix has been shown to have osteoinductive properties in various models and currently is used clinically as bone graft material in posterolateral lumbar spine arthrodesis. Two new formulations of Grafton, one made of flexible sheets (Flex) and the other made in a malleable consistency (Putty), have improved handling characteristics compared with the gel form. In this study, 108 New Zealand white rabbits underwent bilateral posterolateral intertransverse spine arthrodesis at L5-L6 using autogenous iliac crest bone graft alone (control), one of the new forms of demineralized bone matrix (DBM; made from rabbit bone) alone or in combination with autogenous iliac crest bone. Rabbits were killed 6 weeks after surgery. The lumbar spines were excised, and fusion success or failure was determined by manual palpation and radiography. Specimens also were processed for undecalcified histologic analysis. Manual palpation of the harvested lumbar spines revealed that the fusion rates of the Flex-DBM/Auto group (9/9, 100%) and Putty-DBM/Auto group (10/10, 100%) were superior (P < 0.01) to those of the Auto/control group (3/9, 33%). As a stand-alone graft substitute, Flex-DBM performed superiorly with a fusion rate of 11/11 (100%) compared with that of Putty-DBM (10/12, 83%) and Gel-DBM (7/12, 58%). The devitalized version of Flex-DBM had a fusion rate of 4/11 (36%), which was comparable with the devitalized Putty-DBM rate of 4/12 (33%). Both were superior (P < 0.05) to the devitalized Gel-DBM rate of 0/12 (0%). More mature fusions with greater amounts of trabecular bone were present radiographically and histologically in rabbits that received all forms of demineralized bone matrix than in those in which autograft was used. The new flexible sheet and malleable putty forms of demineralized bone matrix were effective as graft extender and graft enhancer in a model of posterolateral lumbar spine fusion. These newer formulations of Grafton appear to have a greater capacity to form bone than the gel form or autogenous bone graft alone in this model.


The effect of biliary decompression on antibiotic biliary excretion.
Raised biliary pressure may affect antibiotic biliary excretion. We evaluated whether biliary decompression for patients with biliary obstruction could improve antibiotic biliary excretion. Eight patients with common bile duct obstruction undergoing endoscopic nasobiliary drainage were evaluated. During endoscopic cannulation, biliary pressure above the obstruction and antibiotic concentrations in the bile and peripheral blood were determined 60 min after the intravenous antibiotic (panipenem) administration. Biliary pressure was initially elevated above normal in all the patients, but normalized after biliary drainage for 5 to 7 days. At the initial endoscopic retrograde cholangiopancreatography, the aspirated bile contained low or undetectable levels of the antibiotic, but the mean bile panipenem concentration and the mean bile/plasma ratio of panipenem concentrations significantly improved after biliary decompression. The results suggest an important role of biliary pressure in determining antibiotic transfer into the bile.


Effects of Dietary Protein and Fat Content on Intrahepatocellular and Intramyocellular Lipids during a 6-Day Hypercaloric, High Sucrose Diet: A Randomized Controlled Trial in Normal Weight Healthy Subjects.
Sucrose overfeeding increases intrahepatocellular (IHCL) and intramyocellular (IMCL) lipid concentrations in healthy subjects. We hypothesized that these effects would be modulated by diet protein/fat content. Twelve healthy men and women were studied on two occasions in a randomized, cross-over trial. On each occasion, they received a 3-day 12% protein weight maintenance diet (WM) followed by a 6-day hypercaloric high sucrose diet (150% energy requirements). On one occasion the hypercaloric diet contained 5% protein and 25% fat (low protein-high fat, LP-HF), on the other occasion it contained 20% protein and 10% fat (high protein-low fat, HP-LF). IHCL and IMCL concentrations (magnetic resonance spectroscopy) and energy expenditure (indirect calorimetry) were measured after WM, and again after HP-LF/LP-HF. IHCL increased from 25.0 ± 3.6 after WM to 147.1 ± 26.9 mmol/kg wet weight (ww) after LP-HF and from 30.3 ± 7.7 to 57.8 ± 14.8 after HP-LF (two-way ANOVA with interaction: p < 0.001 overfeeding x protein/fat content). IMCL increased from 7.1 ± 0.6 to 8.8 ± 0.7 mmol/kg ww after LP-HF and from 6.2 ± 0.6 to 6.9 ± 0.6 after HP-LF, (p < 0.002). These results indicate that liver and muscle fat deposition is enhanced when sucrose overfeeding is associated with a low protein, high fat diet compared to a high protein, low fat diet.


Mutational analysis and characterization of nocardicin C-9' epimerase.
The biosynthetic gene cluster for the nocardicin A producer Nocardia uniformis subsp. tsuyamanensis ATCC 21806 was recently identified. Nocardicin A is the most potent of a series of monocyclic beta-lactam antibiotics produced by this organism. Its activity has been attributed to a syn-configured oxime moiety and a d-homoseryl side chain attached through an unusual ether linkage to the core nocardicin framework. Notably present in the nocardicin biosynthetic gene cluster is nocJ, encoding a protein with sequence similarity to the pyridoxal 5'-phosphate (PLP)-dependent 1-aminocyclopropane-1-carboxylic acid deaminases. Insertional mutagenesis of nocJ abolished nocardicin A production, while the l-homoseryl isomer, isonocardicin A, was still observed. Expression of the disrupted nocJ gene in trans was sufficient to restore production of nocardicin A in the disruption mutant. Heterologous expression, purification, and in vitro characterization of NocJ by UV spectroscopy, cofactor reduction, chiral HPLC analysis of the products and their exchange behavior in deuterium oxide led to confirmation of its role as the PLP-dependent nocardicin C-9' epimerase responsible for interconversion of the nocardicin homoseryl side chain in both nocardicin A with isonocardicin A, and nocardicin C with isonocardicin C. NocJ is the first member of a new class of beta-lactam aminoacyl side chain epimerases, the first two classes being the evolutionarily distinct prokaryotic PLP-dependent isopenicillin N epimerase and the fungal isopenicillin N epimerase two protein system.


T cell costimulation blockade blunts pressure overload-induced heart failure.
Heart failure (HF) is a leading cause of mortality. Inflammation is implicated in HF, yet clinical trials targeting pro-inflammatory cytokines in HF were unsuccessful, possibly due to redundant functions of individual cytokines. Searching for better cardiac inflammation targets, here we link T cells with HF development in a mouse model of pathological cardiac hypertrophy and in human HF patients. T cell costimulation blockade, through FDA-approved rheumatoid arthritis drug abatacept, leads to highly significant delay in progression and decreased severity of cardiac dysfunction in the mouse HF model. The therapeutic effect occurs via inhibition of activation and cardiac infiltration of T cells and macrophages, leading to reduced cardiomyocyte death. Abatacept treatment also induces production of anti-inflammatory cytokine interleukin-10 (IL-10). IL-10-deficient mice are refractive to treatment, while protection could be rescued by transfer of IL-10-sufficient B cells. These results suggest that T cell costimulation blockade might be therapeutically exploited to treat HF.


Automatic pneumonia detection based on ultrasound video analysis.
Pneumonia is a disease which causes high mortality in children under five years old, particularly in developing countries. This paper proposes a novel application of ultrasound video analysis for the detection of pneumonia. This application is based on the processing of small video chunks, in which an image processing algorithm analyzes each frame to get some overall video statistics. Then, based on these quantities, the likeness of presence of pneumonia in the video is determined. The algorithm exploits different geometrical properties of typical anatomical and pathological features that commonly appear in lung sonography and which are already clinically typified in the literature. Our technique has been tested on different transverse thoracic scanning protocols and probe's maneuvers, thus, under a variety of clinical and usage protocols. Then, it can be targeted towards screening applications. We present encouraging results (AUC measure between 0.7851 and 0.9177) based on the analysis of 346 videos with an average duration of eight seconds. The analyzed videos were taken from children who were between three and five years old. Finally, our algorithm can be used directly as a classifier, but we detail how its performance may be enhanced if used as a first stage of a larger pipeline of other complementary pneumonia detection processes.


Genetic constraints on adaptation to a changing environment.
Genetic correlations between traits can constrain responses to natural selection. To what extent such correlations limit adaptation depends on patterns of directional selection. I derive the expected rate of adaptation (or evolvability) under randomly changing selection gradients. When directional selection gradients have an arbitrary covariance matrix, the average rate of adaptation depends on genetic correlations between traits, contrary to the isotropic case investigated in previous studies. Adaptation may be faster on average with more genetic correlation between traits, if these traits are selected to change jointly more often than the average pair of traits. However, natural selection maximizes the long-term fitness of a population, not necessarily its rate of adaptation. I therefore derive the average lag load caused by deviations of the mean phenotype from an optimum, under several forms of environmental changes typically experienced by natural populations, both stochastic and deterministic. Simple formulas are produced for how the G matrix affects long-term fitness in these contexts, and I discuss how their parameters can be estimated empirically.


Isolation and characterization of the CD133+ precursors from the ventricular zone of human fetal brain by magnetic affinity cell sorting.
A fast and effective method to enrich large number of neural precursors from the ventricular zone of human fetus by magnetic affinity cell sorting (MACS) is reported. After incubation with phycoerythrin (PE)-conjugated anti-CD133 antibodies and anti-PE magnetic beads followed by one cycle of MACS, CD133(+) cells were harvested at 85% purity as confirmed by flow-cytometry and immunocytochemistry. In contrast to CD133(-) cells, these CD133(+) cells initiated primary and secondary neurospheres in culture, and the progeny of sorted cells could be differentiated into both neurons and glia, indicating that these highly enriched cells are capable of self-renewal and multi-lineage potential.


Vesico-ureteric reflux.
There is increasing awareness that vesico-ureteric reflux is associated with the occurrence of renal scars. The influence of infection in scar formation is unclear. The way in which corrective surgery may prevent scar development in selected patients is gradually becoming apparent. It would seem probable that surgical correction of vesico-ureteric reflux is indicated only in infants and young children with gross vesico-ureteric reflux.


Soleus H reflex studies in humans at 40-70 bar in helium-oxygen-nitrogen (trimix).
Excitability of the Hoffmann (H) reflex was studied in 6 subjects at pressures of 40 to 70 bar during two trimix exposures (10% N2, 0.5 bar O2, the remainder He; Atlantis II and III simulated dives, Duke University Medical Center). In Atlantis II (12 h to 47 bar, 1 + 3 days to 66 bar), the H reflex recruitment ratio (Hmax/Mmax) was increased in 2 out of 3 subjects and reduced in the 3rd, while the usual facilitatory effect on the H reflex of a hand-grip reinforcement maneuver was reduced in all subjects. These results are similar to those previously seen in He-O2 dives and suggest a slight hyperreflexia mediated by an increase in alpha motoneuron excitability. In Atlantis III (1.5 days to 47 bar, 7 days to 66 bar) H recruitment ratios were normal in 2 subjects, and reduced in 1 subject (P less than 0.001). Early (III) and late (V) inhibitory phases of the H reflex recovery cycle were not significantly altered, while a 50-ms delay in the recovery cycle rebound (phase IV) in 5 out of 6 subjects at 66 bar (P less than 0.01) is attributed to the slowed muscle contraction. However, the increased amplitude of the rebound and the presence of clonic electromyographic potentials in Atlantis III are evidence of stretch receptor hyperexcitability that apparently persisted when H reflex excitability was normal. It is concluded that increased reflex responsiveness at depth is not promoted exclusively by either enhanced alpha motoneuron excitability or stretch receptor sensitivity but is mediated via both systems. Slower compression and the presence of N2 appear to preserve normal spinal inhibitory feedback and alpha motoneuron excitability but do not prevent increases in stretch receptor sensitivity.


Effects of accelerating voltage and specimen thickness on the spatial resolution of transmission electron backscatter diffraction in Cu.
A quantitative approach was proposed to determine the spatial resolution of transmission electron backscatter diffraction (t-EBSD) and to understand the limits of spatial resolution of t-EBSD. In this approach, Cu bicrystals and digital image correlation were employed. The effects of accelerating voltage and specimen thickness on the spatial resolution of t-EBSD were also investigated. t-EBSD specimens with 8μm×10μm dimensions and different thicknesses were prepared using focused ion beam milling. The optimized quality of Kikuchi pattern was achieved at a working distance of 12mm and a tilting angle of 20°. The optimum depth resolution of 34.4nm was observed in the lower surface of a 100nm thick sample at 25kV. Thus, the penetration depth from the upper surface is 65.6nm. The optimum lateral and longitudinal resolutions obtained from a 100nm thick sample at 30kV are 25.2 and 43.4nm, respectively. The spatial resolution of t-EBSD can be enhanced by increasing the accelerating voltage and decreasing the sample thickness.


Role of consolidative surgical therapy in patients with locally advanced or regionally metastatic bladder cancer.
The effect of radical cystectomy and extended pelvic lymph node dissection (RC/PLND) on the survival of patients with locally advanced and/or regionally metastatic bladder cancer is unknown. However, emerging evidence suggests that there may be survival benefit to a subset of select patients with this disease who demonstrate a response to chemotherapy. This article will review the current literature on the role of RC/PLND in the consolidative treatment of locally advanced and regionally metastatic bladder cancer.


Pirinixic acid derivatives as novel dual inhibitors of microsomal prostaglandin E2 synthase-1 and 5-lipoxygenase.
Dual inhibition of the prostaglandin (PG) and leukotriene (LT) biosynthetic pathway is supposed to be superior over single interference, both in terms of efficacy and side effects. Here, we present a novel class of dual microsomal PGE(2) synthase-1/5-lipoxygenase (5-LO) inhibitors based on the structure of pirinixic acid [PA, 2-(4-chloro-6-(2,3-dimethylphenylamino)pyrimidin-2-ylthio)acetic acid, compound 1]. Target-oriented structural modification of 1, particularly alpha substitution with extended n-alkyl or bulky aryl substituents and concomitant replacement of the 2,3-dimethylaniline by a biphenyl-4-yl-methane-amino residue, resulted in potent suppression of mPGES-1 and 5-LO activity, exemplified by 2-(4-(biphenyl-4-ylmethylamino)-6-chloropyrimidin-2-ylthio)octanoic acid (7b, IC(50) = 1.3 and 1 microM, respectively). Select compounds also potently reduced PGE(2) and 5-LO product formation in intact cells. Importantly, inhibition of cyclooxygenases-1/2 was significantly less pronounced. Taken together, these pirinixic acid derivatives constitute a novel class of dual mPGES-1/5-LO inhibitors with a promising pharmacological profile and a potential for therapeutic use.


Emphysematous cystitis: A case report and literature review of 113 cases.
Emphysematous cystitis (EC) is a rare complicated urinary tract infection characterized by gas formation within the wall of the urinary bladder. Although EC has multifactorial etiology, commonly seen in elderly, diabetic and female sex; most of the cases of EC present along with emphysematous pyelonephritis (EPN) and full-blown urosepsis. We present a case of EC with EPN presented with features of sepsis managed conservatively. Furthermore, we reviewed the literature of published cases reports of EC with or without EPN from 1999 to 2019 (20 Years). From eligible 113 case reports, data of clinical presentation, demographic profile, risk factors, diagnostic methods, treatment, and prognosis were analyzed.


Granulocytic sarcoma of the colon and leukemic infiltration of the liver in a patient presenting with hematochezia and jaundice.
Granulocytic sarcoma (GS) is an extramedullary tumor composed of immature cells of the granulocytic series known to occur in patients with myelodysplastic syndrome, chronic myelogenous leukemia, or acute myelogenous leukemia (AML). Involvement of the gastrointestinal tract is relatively rare in GS. We present an extremely rare case of GS of the colon and liver infiltration in a 60-year-old male patient with AML presenting with jaundice and hematochezia and review the literature. It should be kept in mind that hematochezia may be due to colonic involvement of GS besides thrombocytopenia which is usually encountered in patients with AML.


Akt-mTOR signaling is involved in Notch-1-mediated glioma cell survival and proliferation.
Both Notch signaling and Akt-mTOR signaling pathway are involved in glioma cell proliferation and survival. Previous studies have shown that Notch-1 is overexpressed in many glioma cell lines and primary human gliomas. Blocking of Notch signaling pathway can induces glioma cell apoptosis and growth suppression. However, the underlying molecular mechanism is not clear. We report that activation of the Notch pathway by intracellular domain of human Notch-1 (NIC-1) strongly activates Akt and promotes U251 glioma cell proliferation. Knockdown of Notch-1 by RNA interference suppresses Akt activation, reduces glioma cell growth rate and induce cell apoptosis. Following Notch-1 suppression, phosphorylated Akt and its downstream effector mTOR were reduced. Knockdown of Notch-1 also involves down-regulation of anti-apoptotic protein MCL-1, in parallel with activation of apoptotic associate proteins PARP, caspase-9 and caspase-3. Our data demonstrate that Notch-1 can positively regulate Akt-mTOR pathways, which is associated with glioma cell proliferation and apoptosis. This also suggests a molecular mechanism for the inhibitory effect of Notch-1 RNA interference on glioma cell proliferation through Akt-mTOR signaling pathway.


Comparison between Airway Responses to High versus Low Molecular Weight Compounds in Occupational Asthma.
Occupational asthma (OA) is a heterogeneous disease, and the characteristics of the sensitizer responsible for OA may induce different clinical, functional, and biological manifestations. We examined the characteristics of 74 patients with OA induced by low molecular weight compounds (LMWC) or by high molecular weight compounds (HMWC) and diagnosed by specific inhalation challenge (SIC). Patients with OA induced by LMWC had a longer occupational exposure before the beginning of symptoms, a lower sputum eosinophilia, and a higher prevalence of late airway response (LAR), in comparison with patients with OA induced by HMWC. Pulmonary function tended to be poorer and atopy tended to be less frequent in LMWC-induced OA than in HMWC-induced OA. These data confirm and extend previous observations showing that the characteristics of the specific sensitizer inducing OA may determine different clinical, functional, and biological features, probably related to the difference pathogenetic mechanisms underlying these different types of OA.


Crosslinked Microgels as Platform for Hydrolytic Catalysts.
In an effort to develop biomimetic glycosyl transfer catalysts, a polymerization protocol was developed that is applicable to the synthesis of crosslinked microgels via UV-initiated free radical polymerization of miniemulsions at ambient temperature and below. The catalytic activity of the microgels derived from butyl acrylate, EGDMA, and a catalyst-precursor ligand was established using the hydrolysis of 4-methylumbelliferyl β-d-galactopyranoside as a model reaction. The microgel-catalyzed hydrolysis was up to 3 × 105-fold accelerated over the background reaction and notably 38-fold faster than the hydrolysis catalyzed by its low molecular weight analog. Dynamic light scattering analysis demonstrated mean hydrodynamic diameters of the particles between 210 and 280 nm and chemical stability of the particles in aqueous solution between pH 1 and 13. A bell-shaped correlation between catalytic proficiency and material rigidity was observed that peaks at 40 mol % of crosslinking content of the microgel.


Dysregulated signalling pathways in innate immune cells with cystic fibrosis mutations.
Cystic fibrosis (CF) is one of the most common life-limiting recessive genetic disorders in Caucasians, caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR). CF is a multi-organ disease that involves the lungs, pancreas, sweat glands, digestive and reproductive systems and several other tissues. This debilitating condition is associated with recurrent lower respiratory tract bacterial and viral infections, as well as inflammatory complications that may eventually lead to pulmonary failure. Immune cells play a crucial role in protecting the organs against opportunistic infections and also in the regulation of tissue homeostasis. Innate immune cells are generally affected by CFTR mutations in patients with CF, leading to dysregulation of several cellular signalling pathways that are in continuous use by these cells to elicit a proper immune response. There is substantial evidence to show that airway epithelial cells, neutrophils, monocytes and macrophages all contribute to the pathogenesis of CF, underlying the importance of the CFTR in innate immune responses. The goal of this review is to put into context the important role of the CFTR in different innate immune cells and how CFTR dysfunction contributes to the pathogenesis of CF, highlighting several signalling pathways that may be dysregulated in cells with CFTR mutations.


A quantitative analysis of the aging of human glial cells in culture.
The kinetics of aging of normal human diploid brain cells in culture have been determined using the miniclone technique in which cells are cloned in the presence of a large number of other cells. The miniclone technique records the behaviour of every viable cell in the sample, not merely those cells capable of forming visible clones. This technique permits the direct measurement of the reproductive potential of individual cells growing in bulk culture and of the dispersion of the sizes of colonies generated by dividing cells. The fraction of cells that are able to divide declines smoothly and continuously from the beginning of in vitro cultures of human glial cells. There is a broad distribution of colony sizes; even at the earliest passages there are significant numbers of small colonies. With increasing age of the culture there is a shift in the distribution, so that fewer large colonies and more small colonies occur. The distribution of intermitotic times is almost identical in young and middle-aged cultures. Our data seem to exclude quite positively any description in terms of a catastrophe or any abrupt change in the population. On the contrary, the decline in reproductive potential may be described adequately either as a linear change with time, or as predicted by the mortality theory of Shall and Stein (1979), in which the single constant, gamma, describes the change in reproductive potential over the entire lifetime.


Is binding decline the main source of the ageing effect on prospective memory? A ride in a virtual town.
Objective: This study was designed to improve our understanding of prospective memory (PM) changes in ageing, and to identify the cognitive correlates of PM decline, using a virtual environment, to provide a more realistic assessment than traditional laboratory tasks. Design: Thirty-five young and 29 older individuals exposed to a virtual town were asked to recall three event-based intentions with a strong link between prospective and retrospective components, three event-based intentions with a weak link, and three time-based intentions. They also underwent retrospective episodic memory, executive functions, binding in working memory, processing speed, and time estimation assessments. Results: Older individuals recalled fewer intentions than young adults. While age-related PM decline affected the recall of both prospective and retrospective components, the recall of the latter seemed more challenging for older individuals when the link was weak. This PM decline was linked to an age-related decline in the binding process in working memory, as well as in processing speed, executive functioning, and episodic memory, depending on the nature of intentions. Conclusion: PM appears to be sensitive to ageing, even when the device is thought to be ecological. This decline is particularly pronounced when controlled processes are needed.


[Physiopathology of the clinical manifestation of multiple sclerosis. Electrophysiological hypotheses].
Clinical manifestations of multiple sclerosis cannot always be correlated with the type of lesion present. Plaques in certain regions may be clinically silent whereas, inversely, some neurological disorders appear not to be related to a demyelinization process. The pathophysiology of neurological symptoms and signs in multiple sclerosis may therefore be related to two other factors: 1) membrane phenomena observed in demyelinated fibers and leading to partial or complete conduction block, or 2) conduction blocks due to pathological phenomena that fail to provoke anatomical demyelinization. Demyelinization induces a sudden drop in impedance of the axon membrane which is responsible for the conduction block. Redistribution of sodium ionophores in the demyelinated axon enables transmission of certain messages but in a slower and unreliable manner. Sensitivity of demyelinated fibers to temperature and the extracellular ionic environment provides an explanation for the transient, recurrent stereotyped reactions provoked by fever, physical exercise or digestion. In the absence of demyelinization, a conduction block could be the result of either a minimal lesion at the paranodal myelin with denudation of the specialized parts between axon and glia,or of humoral blocking factors--of debatable specificity--or cellular factors that have been demonstrated recently. The existence of such mechanisms suggests the need for revision of the notion of acute episodes of multiple sclerosis. Finally, knowledge of the mechanisms involved in nerve conduction along demyelinated fibers suggests the possibility of therapy to restore conduction of these fibers by acting on extracellular ionic concentrations or directly on membrane ionic canals.


Vibrotactile thresholds in mechanoreceptive afferents innervating the foot pad of the cat. The importance of stimulus frequency and duration.
Vibrotactile thresholds were determined for non-Pacinian (SA and RA) and Pacinian (PC) mechanoreceptive units innervating the foot pad of the cat with vibratory stimuli of varying frequency (20, 80, and 240 Hz) and duration (100, 400, and 800 ms). Both the absolute (1 impulse/stimulus train) and the tuning (1 impulse/cycle) thresholds of SA and RA units were elevated and those of PC units were decreased as a function of increasing vibration frequency. PC units had the lowest thresholds at 80 and at 240 Hz whereas RA units had the lowest thresholds at 20 Hz. Larger atonal intervals (the range between the absolute and the tuning thresholds) were obtained for all unit types with increasing vibration frequency and PC units had the smallest atonal intervals at all frequencies. The absolute thresholds of all unit types were independent of stimulus duration. The tuning thresholds of SA and RA units were elevated with increasing stimulus duration whereas the tuning thresholds of PC units were independent of stimulus duration. Consequently, larger atonal intervals were obtained for RA and SA but not for PC units with increasing stimulus duration. The results indicate that there are both differences (effect of frequency) and similarities (effect of stimulus duration and the width of atonal intervals) in response characteristics of non-Pacinian mechanoreceptors innervating the hairy skin and the glabrous foot pad of the cat.


Single-anastomosis gastric bypass: better, faster, and safer?
Metabolic surgery is evolving and safer, but more effective alternatives are constantly sought for. The single-anastomosis gastric bypass was first described in 2001, and although controversial, this operation is gaining fast in popularity worldwide. This review is aiming at a critical evaluation of the procedure in the light of the literature. The abstracts of 73 articles were reviewed, and after removal of case studies, duplicates, and irrelevant articles, 10 articles remained for closer review. Several thousand of this operation have been performed for more than 15 years. It is claimed to be an easier, safer, faster, and more effective metabolic operation compared to the standard Roux-en-Y gastric bypass. The proponents of this operation also claim that this operation is easier to revise and reverse, leaving more options compared to the situation after standard bypass in case of failure. However, there is much controversy surrounding this method, mainly concerning the possible harmful and in the long-term even carcinogenic effects of biliopancreaticoduodenal reflux in the gastric pouch. The single-anastomosis gastric bypass may be an attractive alternative metabolic operation. However, further studies with robust, long-term follow-up are needed.


Testing time symmetry in time series using data compression dictionaries.
Time symmetry, often called statistical time reversibility, in a dynamical process means that any segment of time-series output has the same probability of occurrence in the process as its time reversal. A technique, based on symbolic dynamics, is proposed to distinguish such symmetrical processes from asymmetrical ones, given a time-series observation of the otherwise unknown process. Because linear stochastic Gaussian processes, and static nonlinear transformations of them, are statistically reversible, but nonlinear dynamics such as dissipative chaos are usually statistically irreversible, a test will separate large classes of hypotheses for the data. A general-purpose and robust statistical test procedure requires adapting to arbitrary dynamics which may have significant time correlation of undetermined form. Given a symbolization of the observed time series, the technology behind adaptive dictionary data compression algorithms offers a suitable estimate of reversibility, as well as a statistical likelihood test. The data compression methods create approximately independent segments permitting a simple and direct null test without resampling or surrogate data. We demonstrate the results on various time-series-reversible and irreversible systems.


Vitamin B12 and folic acid in children with intestinal parasitic infection.
To determine prospectively plasma levels of vitamin B12 and folic acid in children with intestinal parasitic infection before and three months after antiparasitic treatment. 3036 stool samples were collected from 1959 children and 939 cello-tape anal swabs were taken from 688 children for intestinal parasite investigation. Of these, 155 children were identified as having a parasitic infection; however, only 86 were followed up during this study: 26 children with Giardia lamblia infection were treated with tinidazole and metronidazole, pyrantel pamoate was used in the treatment of 40 children with Enterobius vermicularis, and 20 patients infected with Cryptosporidium parvum received only symptomatic treatment. Vitamin B12 and folic acid levels were measured by radioimmunoassay, before and three months after the completion of treatment. Vitamin B12 serum concentrations did not show any significant differences among the three groups. There was a significant increase in vitamin B12 serum concentrations after three months of anti-parasitic treatment (630.57+/-200.97 vs. 667.97+/-181.55 pg/dL, p = 0.002, n = 86). Paired analysis in each group showed only significant increases for vitamin B12 in the Giardia lamblia group and in the Enterobius vermicularis group. No statistically significant differences were found for folic acid serum concentrations before and three months after treatment. Patients with symptomatic infection by Giardia lamblia and Enterobius vermicularis have lower vitamin B12 levels than asymptomatic patients. This could reflect a more affected intestinal mucous. These results could present the opportunity to treat these parasitic infections and to use vitamin B12 supplementation in symptomatic children with Giardia lamblia and Enterobius vermicularis infection.


Fundamental challenges in assessing the impact of research infrastructure.
Clinical research infrastructure is one of the unsung heroes of the scientific response to the current COVID-19 pandemic. The extensive, long-term funding into research support structures, skilled people, and technology allowed the United Kingdom research response to move off the starting blocks at pace by utilizing pre-existing platforms. The increasing focus from funders on evaluating the outcomes and impact of research infrastructure investment requires both a reframing and progression of the current models in order to address the contribution of the underlying support infrastructure. The majority of current evaluation/outcome models focus on a "pipeline" approach using a methodology which follows the traditional research funding route with the addition of quantitative metrics. These models fail to embrace the complexity caused by the interplay of previous investment, the coalescing of project outputs from different funders, the underlying infrastructure investment, and the parallel development across different parts of the system. Research infrastructure is the underpinning foundation of a project-driven research system and requires long-term, sustained funding and capital investment to maintain scientific and technological expertise. Therefore, the short-term focus on quantitative metrics that are easy to collect and interpret and that can be assessed in a roughly 5-year funding cycle needs to be addressed. The significant level of investment in research infrastructure necessitates investment to develop bespoke methodologies that develop fit-for-purpose, longer-term/continual approach(es) to evaluation. Real-world research should reflect real-world evaluation and allow for the accrual of a narrative of value indicators that build a picture of the contribution of infrastructure to research outcomes. The linear approach is not fit for purpose, the research endeavour is a complex, twisted road, and the evaluation approach needs to embrace this complexity through the development of realist approaches and the rapidly evolving data ecosystem. This paper sets out methodological challenges and considers the need to develop bespoke methodological approaches to allow a richer assessment of impact, contribution, attribution, and evaluation of research infrastructure. This paper is the beginning of a conversation that invites the community to "take up the mantle" and tackle the complexity of real-world research translation and evaluation.


Plasma conditions for improved energy coupling into the gain region of the Ni-like Pd transient collisional x-ray laser.
We have directly probed the conditions in which the Ni-like Pd transient collisional x-ray laser is generated and propagates by measuring the near-field image and by utilizing picosecond resolution soft x-ray laser interferometry of the preformed Pd plasma gain medium. The electron density and gain region of the plasma have been determined experimentally and are found to be in good agreement with simulations. We observe a strong dependence of the laser pump-gain medium coupling on the laser pump parameters. The most efficient coupling occurs with the formation of lower density gradients in the preformed plasma and when the duration of the main heating pulse is comparable to the gain lifetime (approximately 10 ps for mid- Z Ni-like schemes). This increases the output intensity by more than an order of magnitude relative to the commonly utilized case where the same pumping energy is delivered within a shorter heating pulse duration (<3 ps) . In contrast, the higher intensity heating pulses are observed to be absorbed at higher electron densities and in regions where steep density gradients limit the effective length of the gain medium.


Electrochemical behavior of titanium exposed to a biofilm supplemented with different sucrose concentrations.
Biofilms can reduce the corrosion resistance of titanium because of the bacterial metabolism of fermentable carbohydrates, including sucrose. However, studies evaluating whether biofilms exposed to higher sucrose concentrations can affect the electrochemical behavior of titanium are lacking. The purpose of this in vitro study was to test the electrochemical behavior of titanium previously exposed to biofilm supplemented with different sucrose concentrations. Streptococcus mutans UA159 biofilms were formed on commercially pure titanium (cpTi) surfaces and supplemented constantly with different sucrose concentrations (0%, 1%, 10%, and 40%) for 7 days (experimental groups) (n=12 per group). CpTi disks without biofilm were used as a control (n=12). The standard electrochemical tests open-circuit potential, electrochemical impedance spectroscopy, and potentiodynamic curve were performed. Data were submitted to ANOVA and the Tukey honestly significant difference (HSD) tests (α=.05). The biofilm exposed to sucrose had an increased biofilm dry weight (P<.05). The polysaccharide amount and the pH drop were higher in the groups exposed to sucrose (P<.05). No difference was noted between the control and experimental groups for the electrochemical properties of cpTi (P>.05). Biofilms exposed to greater carbohydrate concentration did not alter the corrosive behavior of titanium.


18F-FDG PET/CT findings in a patient with blastic plasmacytoid dendritic cell neoplasm and post-transplant lymphoproliferative disorder after hematopoietic stem cell transplantation: a case report.
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is an extremely rare hematopoietic malignancy, which originating from precursors of plasmacytoid dendritic cells. Allogeneic hematopoietic stem cell transplantation (HSCT) is normally considered in the treatment of BPDCN patients to acquire sustained remission. Post-transplant lymphoproliferative disorder (PTLD) is a group of conditions involving abnormal lymphoid cells proliferation in the context of extrinsic immunosuppression after solid organ transplantation (SOT) or HSCT. Herein, we report a patient with BPDCN, who suffered from PTLD after allogeneic HSCT. A 66-year-old man was diagnosed with BPDCN, confirmed by pathologic examination after splenectomy. The post-surgery 18F-fluoro-2-deoxy-D-glucose-positron emission tomography/computed tomography (18F-FDG PET/CT) showed multifocal 18F-FDG avidity in the left cheek, lymph nodes and bone marrow. The patient started chemotherapy, followed by allogeneic HSCT and immunosuppressive therapy. Four months after the HSCT, the patient developed intermittent fever and recurrent lymphadenopathy, accompanied with progressively elevated Epstein-Barr virus (EBV)-DNA both in serum and lymphocytes. 18F-FDG PET/CT was performed again and found multiple new enlarged 18F-FDG-avid lymph nodes, while the previous hypermetabolic lesions all disappeared. The pathology of mesenteric lymph node indicated a monomorphic PTLD (diffuse large B-cell lymphoma). Then the immunosuppressive medications were stopped and two cycles of Rituximab were given, and the follow-up CT scan indicated a complete response. When patients with BPDCN recurred new enlarged lymph nodes after allogeneic HSCT and immunosuppressive therapy, PTLD should be taken into consideration. 18F-FDG PET/CT may provide additional evidence for supporting or refuting the suspicion of PTLD, and suggest lesions accessible for biopsy.


Visualization Techniques of Time-Oriented Data for the Comparison of Single Patients With Multiple Patients or Cohorts: Scoping Review.
Visual analysis and data delivery in the form of visualizations are of great importance in health care, as such forms of presentation can reduce errors and improve care and can also help provide new insights into long-term disease progression. Information visualization and visual analytics also address the complexity of long-term, time-oriented patient data by reducing inherent complexity and facilitating a focus on underlying and hidden patterns. This review aims to provide an overview of visualization techniques for time-oriented data in health care, supporting the comparison of patients. We systematically collected literature and report on the visualization techniques supporting the comparison of time-based data sets of single patients with those of multiple patients or their cohorts and summarized the use of these techniques. This scoping review used the PRISMA-ScR (Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews) checklist. After all collected articles were screened by 16 reviewers according to the criteria, 6 reviewers extracted the set of variables under investigation. The characteristics of these variables were based on existing taxonomies or identified through open coding. Of the 249 screened articles, we identified 22 (8.8%) that fit all criteria and reviewed them in depth. We collected and synthesized findings from these articles for medical aspects such as medical context, medical objective, and medical data type, as well as for the core investigated aspects of visualization techniques, interaction techniques, and supported tasks. The extracted articles were published between 2003 and 2019 and were mostly situated in clinical research. These systems used a wide range of visualization techniques, most frequently showing changes over time. Timelines and temporal line charts occurred 8 times each, followed by histograms with 7 occurrences and scatterplots with 5 occurrences. We report on the findings quantitatively through visual summarization, as well as qualitatively. The articles under review in general mitigated complexity through visualization and supported diverse medical objectives. We identified 3 distinct patient entities: single patients, multiple patients, and cohorts. Cohorts were typically visualized in condensed form, either through prior data aggregation or through visual summarization, whereas visualization of individual patients often contained finer details. All the systems provided mechanisms for viewing and comparing patient data. However, explicitly comparing a single patient with multiple patients or a cohort was supported only by a few systems. These systems mainly use basic visualization techniques, with some using novel visualizations tailored to a specific task. Overall, we found the visual comparison of measurements between single and multiple patients or cohorts to be underdeveloped, and we argue for further research in a systematic review, as well as the usefulness of a design space.


Impact of risk-based partial vaccination on clinical incidence and seroprevalence of foot and mouth disease in Lao PDR.
Foot and mouth disease (FMD) is endemic in the Lao People's Democratic Republic (PDR) and it causes economic loss to smallholder husbandry systems. An intervention programme based on a risk-based partial vaccination strategy was implemented in three provinces of Lao PDR (Champasak, Savannakhet and Xiangkhouang) to immunise domestic cattle and buffalo during 2016-2020. Two cross-sectional surveys were conducted in 2016/17 and 2020 to evaluate the impact of the vaccination programme on the prevalence of FMD virus exposure and clinical incidence of the disease. A total of 212 villages were visited during the two surveys, collecting 1609 household-level questionnaire results and 5931 blood samples of domestic cattle and buffalo. Blood samples were tested for the presence of antibodies to the non-structural proteins of FMD virus, and seroprevalence of 42.5 and 47.5% in 2016/17 and 2020, respectively were found. Multivariable regression analysis indicated that the efficacy of the FMD vaccination programme for reducing FMD virus circulation varied by province. In general, the incidence of clinical FMD increased toward the end of the 5-year intervention period, coinciding with a reduction of vaccine coverage in the last 2 years of the period. The findings suggest that the risk-based vaccination strategy achieved a marginally protective effect against the circulation of FMD virus with the possible limiting factors being operational constraints of public veterinary services, lack of farmers' compliance and unsustainable funding. We conclude that consistent resource availability and higher vaccination coverage is required to successfully control FMD with a risk-based vaccination strategy in Lao PDR.


Thrombolysis in thromboembolic diseases.
Thrombolysis in many manifestations of thromboembolic disease offers a valuable alternative to surgery. However, as thrombolysis is always associated with a bleeding hazard (though low) one should always weigh the risks against the expected benefits when the decision for or against this therapeutic option is made. Furthermore, in selecting the appropriate thrombolytic agent, one should be led by the urgency of reperfusion to maintain organ function. If one decides on an aggressive, high-dose, brief-duration regimen, reperfusion may be achieved more rapidly but may be incomplete in the majority of cases. On the other hand, by selecting an intermediate- or long-duration, low-dose regimen, reperfusion may happen too late to improve the patient's prognosis. Above all, one should keep in mind that the hazard of serious bleeding constantly increases with duration of thrombolysis. No matter which strategy is regarded as the best to resolve a clot in a particular patient with a particular type of thromboembolic disease, thrombolysis should be accompanied by high doses of i.v. heparin. Finally, if bleeding occurs in spite of all precautions taken, the new generation of fibrin-specific thrombolytic agents offers the advantage of short half-lives. In addition--in contrast to streptokinase--the hemostatic defect that they cause may be rapidly reversed by the infusion of antagonist drugs such as aprotinin, tranexamic acid, or epsilon-aminocaproic acid. This adds to the clinical safety profile of these thrombolytic agents.


Cable grafting of the spinal accessory nerve after radical neck dissection.
From January 1981 through March 1996, 20 patients with head and neck cancer underwent radical neck dissection with sacrifice of the spinal accessory nerve and immediate reconstruction of the nerve using a microsurgical technique and a cable graft of the great auricular nerve. Postoperative shoulder function was assessed via a subjective questionnaire, objective strength testing, and/or postoperative electromyography. The latter was used to evaluate for the presence and amplitude of voluntary motor potentials, the presence of fibrillation potentials, and nerve conduction latency. The group of patients who underwent cable grafting of the spinal accessory nerve was compared with a group of patients who underwent modified radical neck dissection with preservation of the spinal accessory nerve and with another group of patients who underwent a classic neck dissection with sacrifice of the spinal accessory nerve and no reconstruction. In terms of shoulder function, the group of patients in whom the spinal accessory nerve was reconstructed occupied an intermediate position; ie, their postoperative shoulder function was better than that of the patients who underwent radical neck dissection without reconstruction but not as good as that of the patients who underwent modified neck dissection with preservation of the spinal accessory nerve. Cable grafting of the spinal accessory nerve that has been sacrificed during radical neck dissection results in improved shoulder function in the postoperative period.


[Daily approaches of sexuality].
Dealing with STD goes beyond the simple notion of "disease" and implies for the doctor to pay attention to the possible consequences on his patients' sexuality and couple relationship. Indeed, feelings of shame, culpability, fear in the present context of the AIDS epidemic can develop very fast as well as suspicion and loss of trust considering the problem of transmission. Couple's crisis and sexual problems could result from it. One of the most important elements of STD prevention, in particular AIDS is communication around sexuality. The doctor's role is very important as a mediator, he has to treat the dimensions of Safer Sex and use of condoms taking into account the individual needs (and not only the technical aspects) because the sexologic material does not uncover by itself.


The protective effect of melatonin on brain ischemia and reperfusion in rats and humans: In vivo assessment and a randomized controlled trial.
Carotid endarterectomy (CEA) is the treatment of choice for carotid stenosis. Some patients develop ischemia and reperfusion (I/R) injury after CEA. This study was designed to investigate the neuroprotective effects of melatonin on I/R injury in both rats and humans. To this end, 36 male rats were evaluated, and a double-blind randomized controlled trial (RCT) including 60 patients was performed. A rat model of middle cerebral artery occlusion was used to mimic cerebral I/R. After 2 hour of occlusion and 24 hour of reperfusion, blood samples and brain tissues were harvested for further assessments. Compared with the vehicle treatment, melatonin decreased the expression of nuclear factor κ light-chain-enhancer of activated B cells (NF-κB) and S100 calcium-binding protein β (S100β) (P < 0.05) and markedly increased the expression of nuclear erythroid 2-related factor 2 (Nrf2), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) (P < 0.05). The participants in the RCT took 6 mg/d melatonin orally from 3 days before surgery to 3 days after surgery. Blood samples were drawn at the following times: baseline; pre-anesthesia; carotid reconstruction completion; and 6, 24, and 72 hour after CEA. Compared with the oral placebo treatment, melatonin decreased the expression of NF-κB, tumor necrosis factor-α, interleukin-6 (IL-6), and S100β (P < 0.05) and increased the expression of Nrf2, SOD, CAT, and GPx (P < 0.05) in patients after CEA. Our findings suggested that melatonin could ameliorate brain I/R injury after CEA and that this outcome was essentially due to the antioxidant and anti-inflammatory effects of melatonin.


Loss of the flagellum happened only once in the fungal lineage: phylogenetic structure of kingdom Fungi inferred from RNA polymerase II subunit genes.
At present, there is not a widely accepted consensus view regarding the phylogenetic structure of kingdom Fungi although two major phyla, Ascomycota and Basidiomycota, are clearly delineated. Regarding the lower fungi, Zygomycota and Chytridiomycota, a variety of proposals have been advanced. Microsporidia may or may not be fungi; the Glomales (vesicular-arbuscular mycorrhizal fungi) may or may not constitute a fifth fungal phylum, and the loss of the flagellum may have occurred either once or multiple times during fungal evolution. All of these issues are capable of being resolved by a molecular phylogenetic analysis which achieves strong statistical support for major branches. To date, no fungal phylogeny based upon molecular characters has satisfied this criterion. Using the translated amino acid sequences of the RPB1 and RPB2 genes, we have inferred a fungal phylogeny that consists largely of well-supported monophyletic phyla. Our major results, each with significant statistical support, are: (1) Microsporidia are sister to kingdom Fungi and are not members of Zygomycota; that is, Microsporidia and fungi originated from a common ancestor. (2) Chytridiomycota, the only fungal phylum having a developmental stage with a flagellum, is paraphyletic and is the basal lineage. (3) Zygomycota is monophyletic based upon sampling of Trichomycetes, Zygomycetes, and Glomales. (4) Zygomycota, Basidiomycota, and Ascomycota form a monophyletic group separate from Chytridiomycota. (5) Basidiomycota and Ascomycota are monophyletic sister groups. In general, this paper highlights the evolutionary position and significance of the lower fungi (Zygomycota and Chytridiomycota). Our results suggest that loss of the flagellum happened only once during early stages of fungal evolution; consequently, the majority of fungi, unlike plants and animals, are nonflagellated. The phylogeny we infer from gene sequences is the first one that is congruent with the widely accepted morphology-based classification of Fungi. We find that, contrary to what has been published elsewhere, the four morphologically defined phyla (Ascomycota, Basidiomycota, Zygomycota and Chytridiomycota) do not overlap with one another. Microsporidia are not included within kingdom Fungi; rather they are a sister-group to the Fungi. Our study demonstrates the applicability of protein sequences from large, slowly-evolving genes to the derivation of well-resolved and highly supported phylogenies across long evolutionary distances.


Identifying relevant asymmetry features of EEG for emotion processing.
The left and right hemispheres of the brain process emotion differently. Neuroscientists have proposed two models to explain this difference. The first model states that the right hemisphere is dominant over the left to process all emotions. In contrast, the second model states that the left hemisphere processes positive emotions, whereas the right hemisphere processes negative emotions. Previous studies have used these asymmetry models to enhance the classification of emotions in machine learning models. However, little research has been conducted to explore how machine learning models can help identify associations between hemisphere asymmetries and emotion processing. To address this gap, we conducted two experiments using a subject-independent approach to explore how the asymmetry of the brain hemispheres is involved in processing happiness, sadness, fear, and neutral emotions. We analyzed electroencephalogram (EEG) signals from 15 subjects collected while they watched video clips evoking these four emotions. We derived asymmetry features from the recorded EEG signals by calculating the log ratio between the relative energy of symmetrical left and right nodes. Using the asymmetry features, we trained four binary logistic regressions, one for each emotion, to identify which features were more relevant to the predictions. The average AUC-ROC across the 15 subjects was 56.2, 54.6, 51.6, and 58.4% for neutral, sad, fear, and happy, respectively. We validated these results with an independent dataset, achieving comparable AUC-ROC values. Our results showed that brain lateralization was observed primarily in the alpha frequency bands, whereas for the other frequency bands, both hemispheres were involved in emotion processing. Furthermore, the logistic regression analysis indicated that the gamma and alpha bands were the most relevant for predicting emotional states, particularly for the lateral frontal, parietal, and temporal EEG pairs, such as FT7-FT8, T7-T8, and TP7-TP8. These findings provide valuable insights into which brain areas and frequency bands need to be considered when developing predictive models for emotion recognition.


Analysis of the Kinetics of the immune response by means of ionizing radiation.
The inhibitory kinetics of the primary immune response was studied by means of lethal irradiation. CBA mice were immunized with sheep red blood cells and 48, 72 and 96 hours later, they were irradiated with 750R. Afterwards, the number of direct plaque-forming cells was determined in their spleens during 58 to 66 hours at 4-hour intervals. After irradiation at 48 and 72 hours of the immune response, a plateau of different duration was obtained: 48 hours in the former and 24 hours in the latter case. In both cases, reduction in the PFC number began at about 96 hours of the resonse. The PFC reduction curves in these two series of experiments and those obtained 96 hours after irradiation and in control, non-irradiated mice were parallel. The mean half-time of the PFC number reduction as determined by these curves was 6 hours. The time of functioning of PFC in the spleen is discussed with respect to recruitment of proliferating precursors.


Overexpression of the catalytic subunit of protein phosphatase 2A impairs cardiac function.
Reversible protein phosphorylation is an essential regulatory mechanism in many cellular functions. In contrast to protein kinases, the role and regulation of protein phosphatases has remained ambiguous. To address this issue, we generated transgenic mice that overexpress the catalytic subunit alpha of protein phosphatase 2A (PP2A) (PP2Acalpha) in the heart driven by the alpha-myosin heavy chain promoter. Overexpression of the PP2Acalpha gene in the heart led to increased levels of the transgene both at RNA and protein levels. This was accompanied by a significant increase of PP2A enzyme activity in the myocardium. Morphological analysis revealed isles of necrosis and fibrosis. The phosphorylation state of phospholamban, troponin inhibitor, and eukaryotic elongation factor 2 was reduced significantly. The expression of junctional (calsequestrin) and free SR proteins (SERCA and phospholamban) was not altered. Whereas no increase in morbidity or mortality was noted, transgenic mice developed cardiac hypertrophy and reduced contractility of the heart, as well as cardiac dilatation as shown by biplane echocardiography. Taken together, these findings are indicative of the fundamental role of PP2A in cardiac function and imply that disturbances in protein phosphatases expression and activity may cause or aggravate the course of cardiac diseases.


Serum potassium and aldosterone levels in gossypol-treated rats.
Male rats fed with 30 mg/kg gossypol for 2 or 4 weeks exhibited a decrease in growth but showed no change from control in serum potassium levels. Serum aldosterone levels remained unchanged, but serum testosterone levels were significantly reduced after 4 weeks of treatment. Serum T3 was significantly increased after 2 or 4 weeks of treatment, while serum T4 was not different from control. It is concluded that gossypol at a dose that is sufficient to produce sterility in rats did not change serum potassium and aldosterone levels.


Associations of Traumatic Injury with Abnormal Glucose Metabolism: A Population-Based Prospective Cohort Study.
Empirical data on the association between traumatic injury and abnormal glucose metabolism risk is limited. This study aimed to investigate the association between traumatic injury and abnormal glucose metabolism. This study included 153,162 participants in the Kailuan Study from 2006 to 2013. Participants with abnormal glucose metabolism at baseline were excluded. All participants were monitored every two years until December 31, 2019. During follow-up, 1915 subjects with a first traumatic injury (defined as a physical injury caused by an external force) were identified. For each subject with traumatic injury, one control subject was randomly selected and matched for age (± 3 years) and sex. A total of 3830 subjects were included in the final analysis. Cox proportional hazards models were used to examine the association between traumatic injury and the subsequent risk of abnormal glucose metabolism. During a median follow-up of 6.91 (3.57-9.41) years, 990 abnormal glucose metabolism events occurred. After adjustment for demographics, lifestyle behaviors, and traditional risk factors, those who had traumatic injury compared to controls were 32% more likely to develop any abnormal glucose metabolism (hazard ratio [HR] 1.32; 95% confidence interval [CI]1.16-1.49), including impaired fasting glucose (IFG) (HR 1.29; 95% CI 1.12-1.48) and diabetes (HR 1.37; 95% CI 1.10-1.70). The risks for abnormal glucose metabolism, IFG, and diabetes in subjects with moderate-severe injury were higher than in subjects with mild injury for the 1-year follow-up period, while the association was not significantly different by injury severity for the whole follow-up period. Traumatic injury was associated with an increased risk of abnormal glucose metabolism. However, the risks of outcome events decreased as the follow-up period extended. Improved short- and long-term prevention and management strategies for controlling glucose are needed for individuals with traumatic injury.


Near-fatal misuse of medical tape around an endotracheal tube connector during inhalation anesthesia in a horse.
A 7-year-old gelded Irish sports horse weighing 650 kg was anesthetized on 2 consecutive days for lavage of a septic right radio-carpal joint. On both occasions the endotracheal tube connector, which had been bound in medical tape to produce an airtight seal, functioned as a unidirectional valve during mechanical ventilation, retarding expiration, imposing positive end expiratory pressure (PEEP), and probably continuous positive airway pressure (CPAP). The equipment dysfunction was not identified on either occasion despite close inspection prompted by progressive increases in airway pressure and thoracic distension. Whilst the PEEP and CPAP exerted unexpectedly modest cardiovascular effects and the horse recovered uneventfully on both occasions, the improvisation may have proven fatal in a higher-risk subject. A 7-year-old gelded Irish sports horse weighing 650 kg was anesthetized on 2 consecutive days for lavage of a septic right radio-carpal joint. On both occasions the endotracheal tube connector, which had been bound in medical tape to produce an airtight seal, functioned as a unidirectional valve during mechanical ventilation, retarding expiration, imposing positive end expiratory pressure (PEEP), and probably continuous positive airway pressure (CPAP). The equipment dysfunction was not identified on either occasion despite close inspection prompted by progressive increases in airway pressure and thoracic distension. Whilst the PEEP and CPAP exerted unexpectedly modest cardiovascular effects and the horse recovered uneventfully on both occasions, the improvisation may have proven fatal in a higher-risk subject.


Influence of carbon dioxide plasma treatment on the dry adhesion of vertical aligned carbon nanotube arrays.
Vertical aligned carbon nanotube (VACNT) arrays used as dry adhesive materials have broad prospects in the applied fields of space, medicine and electronics. The adhesion of VACNT arrays is believed to be related not only to the nano-array structure and a clean surface, but also to chemical composition. Here, radio-frequency (RF) carbon dioxide (CO2) plasma treatment is introduced as an effective method for purifying and functionalizing the surface to improve the dry adhesive performance of VACNT arrays. Scanning electron microscopy showed that the VACNT arrays retained the alignment architecture with minimal damage at low power. X-ray photoelectron spectroscopy and contact angle tests revealed that the content of non-polar components (C = C bonds) decreased after treatment, while the content of polar groups (C = O and O-C = O bonds) increased, which changed the surface polarity of the VACNT arrays. Raman analyses and transmission electron microscopy demonstrated that amorphous carbon can be selectively removed with increasing time (0-18 min), but was continuously generated with increasing power (30-90 W). The best adhesive strength of 18 N cm-2 (increased by 39%) was obtained after CO2 plasma treating for 10 min at 30 W power, which was attributed to the combined action of purification and polarization.


Inhibitor and ion binding sites on the gastric H,K-ATPase.
The gastric H,K-ATPase catalyzes electroneutral exchange of H(+) for K(+) as a function of enzyme phosphorylation and dephosphorylation during transition between E(1)/E(1)-P (ion site in) and E(2)-P/E(2) (ion site out) conformations. Here we present homology modeling of the H,K-ATPase in the E(2)-P conformation as a means of predicting the interaction of the enzyme with two known classes of specific inhibitors. All known proton pump inhibitors, PPIs, form a disulfide bond with cysteine 813 that is accessible from the luminal surface. This allows allocation of the binding site to a luminal vestibule adjacent to Cys813 enclosed by part of TM4 and the loop between TM5 and TM6. K(+) competitive imidazo-1,2alpha-pyridines also bind to the luminal surface of the E(2)-P conformation, and their binding excludes PPI reaction. This overlap of the binding sites of the two classes of inhibitors combined with the results of site-directed mutagenesis and cysteine cross-linking allowed preliminary assignment of a docking mode for these reversible compounds in a position close to Glu795 that accounts for the detailed structure/activity relationships known for these compounds. The new E(2)-P model is able to assign a possible mechanism for acid secretion by this P(2)-type ATPase. Several ion binding side chains identified in the sr Ca-ATPase by crystallography are conserved in the Na,K- and H,K-ATPases. Poised in the middle of these, the H,K-ATPase substitutes lysine in place of a serine implicated in K(+) binding in the Na,K-ATPase. Molecular models for hydronium binding to E(1) versus E(2)-P predict outward displacement of the hydronium bound between Asp824, Glu820, and Glu795 by the R-NH(3)(+) of Lys791 during the conformational transition from E(1)P and E(2)P. The site for luminal K(+) binding at low pH is proposed to be between carbonyl oxygens in the nonhelical part of the fourth membrane span and carboxyl oxygens of Glu795 and Glu820. This site of K(+) binding is predicted to destabilize hydrogen bonds between these carboxylates and the -NH(3)(+) group of Lys791, allowing the Lys791 side chain to return to its E(1) position.


Warming can destabilize predator-prey interactions by shifting the functional response from Type III to Type II.
The potential for climate change and temperature shifts to affect community stability remains relatively unknown. One mechanism by which temperature may affect stability is by altering trophic interactions. The functional response quantifies the per capita resource consumption by the consumer as a function of resource abundance and is a suitable framework for the description of nonlinear trophic interactions. We studied the effect of temperature on a ciliate predator-prey pair (Spathidium sp. and Dexiostoma campylum) by estimating warming effects on the functional response and on the associated conversion efficiency of the predator. We recorded prey and predator dynamics over 24 hr and at three temperature levels (15, 20 and 25°C). To these data, we fitted a population dynamic model including the predator functional response, such that the functional response parameters (space clearance rate, handling time and density dependence of space clearance rate) were estimated for each temperature separately. To evaluate the ecological significance of temperature effects on the functional response parameters, we simulated predator-prey population dynamics. We considered the predator-prey system to be destabilized, if the prey was driven extinct by the predator. Effects of increased temperature included a transition of the functional response from a Type III to a Type II and an increase of the conversion efficiency of the predator. The simulated population dynamics showed a destabilization of the system with warming, with greater risk of prey extinction at higher temperatures likely caused by the transition from a Type III to a Type II functional response. Warming-induced shifts from a Type III to II are not commonly considered in modelling studies that investigate how population dynamics respond to warming. Future studies should investigate the mechanism and generality of the effect we observed and simulate temperature effects in complex food webs including shifts in the type of the functional response as well as consider the possibility of a temperature-dependent conversion efficiency.


Effects of exogenous and endogenous cues on attentional orienting in deaf adults.
Adults who are deaf have been shown to have better visual attentional orienting than those with typical hearing, especially when the target is located in the periphery of the visual field. However, most studies in this population have assessed exogenous visual attention orienting (bottom-up processing of external cues) rather than endogenous visual attention orienting (top-down processing of internal cues). We used a target detection task to assess both types of visual attention orienting. A modified cue-target paradigm was adopted to assess the facilitation effects of exogenous and endogenous cues during short and long inter-stimulus intervals (ISI), using a 2 (Group: deaf/typically hearing) * 2 (Location: central/peripheral) * 2 (Cue Type: exogenous/endogenous) mixed factorial design. ANOVAs showed that both exogenous cues and endogenous cues can facilitate deaf adults' visual attentional orienting, and the facilitation effect of exogenous cues on attention orienting was significantly stronger for deaf participants than hearing participants. When the ISI was long, the effect was significantly stronger when the exogenous cue appeared in the periphery of the visual field. In the periphery, deaf adults benefited most from exogenous cues, whereas hearing adults benefited most from endogenous cues. The results suggest that not only exogenous cues but also endogenous cues can facilitate deaf adults' visual attentional orienting. However, the effect of exogenous cues appears to be greater, especially when the stimulus appears in the peripheral visual field.


Color vision: "OH-site" rule for seeing red and green.
Eyes gather information, and color forms an extremely important component of the information, more so in the case of animals to forage and navigate within their immediate environment. By using the ONIOM (QM/MM) (ONIOM = our own N-layer integrated molecular orbital plus molecular mechanics) method, we report a comprehensive theoretical analysis of the structure and molecular mechanism of spectral tuning of monkey red- and green-sensitive visual pigments. We show that interaction of retinal with three hydroxyl-bearing amino acids near the β-ionone ring part of the retinal in opsin, A164S, F261Y, and A269T, increases the electron delocalization, decreases the bond length alternation, and leads to variation in the wavelength of maximal absorbance of the retinal in the red- and green-sensitive visual pigments. On the basis of the analysis, we propose the "OH-site" rule for seeing red and green. This rule is also shown to account for the spectral shifts obtained from hydroxyl-bearing amino acids near the Schiff base in different visual pigments: at site 292 (A292S, A292Y, and A292T) in bovine and at site 111 (Y111) in squid opsins. Therefore, the OH-site rule is shown to be site-specific and not pigment-specific and thus can be used for tracking spectral shifts in any visual pigment.


Electroperturbation of the human skin barrier in vitro: II. Effects on stratum corneum lipid ordering and ultrastructure.
In transdermal iontophoresis, drugs can be driven across the skin by electrorepulsion, but their transport can also be enhanced by electrical perturbation of the skin barrier. Our objective was to study perturbing effects of electrical current on human stratum corneum lipid fine structure combining techniques including freeze-fracture electron microscopy. Human stratum corneum was subjected to pulsed constant currents, varying from 0.013-13 mA.cm-2. The voltage across the stratum corneum was high-frequency-sampled and s.c. impedence values derived from it. Upon termination of the current, skin samples were rapidly frozen and processed for freeze-fracture electron microscopy or subjected to X-ray diffraction analysis. Initially a rapid decrease of the resistance and, overall, a rapid increase of the capacitances was observed; generally, these effects became more pronounced with increasing current density. Wide- and small-angle X-ray diffractograms of human stratum corneum exposed for 1 h to the highest current indicated a disordering of both the lateral packaging arrangement and long-range lamellar stacking of the intercellular lipids of stratum corneum. Furthermore, an increase in the stratum corneum hydration level as a result of electrical current application was observed. On electron micrographs of freeze-fracture replicas of human stratum corneum, exposed for 1 h to current densities between 0.013 and 13 mA.cm-2, perturbations of the intercellular lipid structure were observed in accordance with the results of X-ray diffraction; these perturbations aggravated with increasing current density. Together, the data suggest that both the lateral and the longitudinal disordering of the intercellular lipids observed with X-ray diffraction may be responsible for the appearance of perturbed structures observed with freeze-fracture electron microscopy. The lipid disordering may be due to polarization of the lipid head groups induced by the electrical field, followed by mutual repulsion.


[Morphological observation of cone biopsy and hysterectomy specimens of high-grade cervical intraepithelial neoplasia].
To study the histopathological features, growth pattern and distribution on the uterine cervix of high-grade cervical intraepithelial neoplasia (CIN). Cone biopsy and hysterectomy specimens of 592 women with high-grade CIN (CIN II or III) were reviewed morphologically. Of all cases, the gross appearance of the external os was recorded, and the frequency of cervical quadrants involved in high-grade CIN was assessed. Two hundred specimens were selected randomly to observe micrographical changes, including the extent, depth, growth pattern and cell types of high-grade CIN and the alteration of stroma surrounding CIN. Of 592 specimens, external os showed smoothness in 46 cases (7.8%), mild erosion in 151 cases (25.5%), moderate or severe erosion in 395 cases (66.7%), hypertrophy in 48 cases (8.1%), and neoplasm in 4 cases (0.7%). High-grade CIN involvement was equally frequent on each quadrant of cervix (P>0.05). Of 200 cases of high-grade CIN reviewed micrographically, the depth of the deepest involved crypt was less than 3 mm in 188 specimens (94.0%), between 3 to 5 mm in 12 specimens (6.0%), and more than 5 mm in none. High-grade CIN with isolated endocervical crypt involvement was detected in 77 cases (38.5%), multiple crypt involvement in 143 cases (71.5%), and confluent crypt involvement in 73 cases (36.5%). High-grade CIN showed explanate in 81.0% (162/200) of cases, endophytic in 49.0% (98/200) of cases, and exophytic in 6. 5% (13/200) of cases. Abrupt transition between CIN and normal epithelia were found in 167 specimens (83.5%), whereas gradual transition in 66 specimens (33.0%). A variety of cell types, but basaloid type predominantly, were observed in high-grade CIN. In 28.5%-45.0% of cases, the stroma surroundin CIN displayed oedema, desmoplastic and lymphocytic responses. Our data suggest that gross examination is of little importance for diagnoses of CIN. CIN trend to spread horizontally and along the endocervical crypts, and are mainly distributed to a depth less than 3 mm. These data also suggest that there are a variety of cell types of CIN, and stroma responses to CIN may exist in part of high-grade CIN.