Serum immunoglobulin free light-chain measurement in primary amyloidosis: prognostic value and correlations with clinical features.
Immunoglobulin free light chains (FLCs) are the precursors of amyloid fibrils in primary amyloidosis (AL). We studied the relationship between FLC levels and clinical features in 730 patients with newly diagnosed AL. The plasma cell clone was λ in 72% patients, and κ in 28% patients. κ-AL had more GI tract and liver involvement, where as renal involvement was more with λ-AL. While the overall survival (OS) was similar for κ and λ-AL, the median OS for those without an identifiable serum heavy chain was significantly shorter (12.6 vs 29.9 months; P = .02). The OS was shorter among those with a higher dFLC (involved FLC-uninvolved FLC; κ > 29.4 mg/dL or λ > 18.2 mg/dL using median for cutoff); 10.9 vs 37.1 months; P < .001. In multivariate analysis, dFLC was independent of other prognostic factors. The type of light chain impacts the spectrum of organ involvement and the FLC burden correlates with survival in AL.


Profiling stem cell states in three-dimensional biomaterial niches using high content image informatics.
A predictive framework for the evolution of stem cell biology in 3-D is currently lacking. In this study we propose deep image informatics of the nuclear biology of stem cells to elucidate how 3-D biomaterials steer stem cell lineage phenotypes. The approach is based on high content imaging informatics to capture minute variations in the 3-D spatial organization of splicing factor SC-35 in the nucleoplasm as a marker to classify emergent cell phenotypes of human mesenchymal stem cells (hMSCs). The cells were cultured in varied 3-D culture systems including hydrogels, electrospun mats and salt leached scaffolds. The approach encompasses high resolution 3-D imaging of SC-35 domains and high content image analysis (HCIA) to compute quantitative 3-D nuclear metrics for SC-35 organization in single cells in concert with machine learning approaches to construct a predictive cell-state classification model. Our findings indicate that hMSCs cultured in collagen hydrogels and induced to differentiate into osteogenic or adipogenic lineages could be classified into the three lineages (stem, adipogenic, osteogenic) with ⩾80% precision and sensitivity, within 72h. Using this framework, the augmentation of osteogenesis by scaffold design exerted by porogen leached scaffolds was also profiled within 72h with ∼80% high sensitivity. Furthermore, by employing 3-D SC-35 organizational metrics, differential osteogenesis induced by novel electrospun fibrous polymer mats incorporating decellularized matrix could also be elucidated and predictably modeled at just 3days with high precision. We demonstrate that 3-D SC-35 organizational metrics can be applied to model the stem cell state in 3-D scaffolds. We propose that this methodology can robustly discern minute changes in stem cell states within complex 3-D architectures and map single cell biological readouts that are critical to assessing population level cell heterogeneity. The sustained development and validation of bioactive materials relies on technologies that can sensitively discern cell response dynamics to biomaterials, while capturing cell-to-cell heterogeneity and preserving cellular native phenotypes. In this study, we illustrate the application of a novel high content image informatics platform to classify emergent human mesenchymal stem cell (hMSC) phenotypes in a diverse range of 3-D biomaterial scaffolds with high sensitivity and precision, and track cell responses to varied external stimuli. A major in silico innovation is the proposed image profiling technology based on unique three dimensional textural signatures of a mechanoreporter protein within the nuclei of stem cells cultured in 3-D scaffolds. This technology will accelerate the pace of high-fidelity biomaterial screening.


Benzyl Furanones and Pyrones from the Marine-Derived Fungus Aspergillus terreus Induced by Chemical Epigenetic Modification.
Chemical epigenetic modification on a marine-derived fungus Aspergillus terreus RA2905 using a histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), resulted in a significantly changed metabolic profile. A chemical investigation of its ethyl acetate (EtOAc) extract led to the isolation of a racemate of benzyl furanone racemate (±)-1, which further separated chirally as a pair of new enantiomers, (+)- and (-)-asperfuranone (1), together with two new benzyl pyrones, asperpyranones A (2) and B (3). Their structures were elucidated by analysis of the comprehensive spectroscopic data, including one-dimensional (1D) and two-dimensional (2D) NMR, and HRESIMS. The absolute configurations were determined by electronic circular dichroism (ECD) calculation and single-crystal X-ray crystallographic experiment. The structures with benzyl furanone or benzyl pyrone skeletons were discovered from natural products for the first time. Compounds (±)-1, (+)-1, (-)-1, and 2 displayed the antifungal activities against Candida albicans with MIC values of 32, 16, 64, and 64 μg/mL and PTP1B inhibitory activities with the IC50 values of 45.79, 17.32, 35.50, and 42.32 μM, respectively. Compound 2 exhibited antibacterial activity against Pseudomonas aeruginosa with the MIC value of 32 μg/mL.


Learning Words While Listening to Syllables: Electrophysiological Correlates of Statistical Learning in Children and Adults.
From an early age, exposure to a spoken language has allowed us to implicitly capture the structure underlying the succession of speech sounds in that language and to segment it into meaningful units (words). Statistical learning (SL), the ability to pick up patterns in the sensory environment without intention or reinforcement, is thus assumed to play a central role in the acquisition of the rule-governed aspects of language, including the discovery of word boundaries in the continuous acoustic stream. Although extensive evidence has been gathered from artificial languages experiments showing that children and adults are able to track the regularities embedded in the auditory input, as the probability of one syllable to follow another syllable in the speech stream, the developmental trajectory of this ability remains controversial. In this work, we have collected Event-Related Potentials (ERPs) while 5-year-old children and young adults (university students) were exposed to a speech stream made of the repetition of eight three-syllable nonsense words presenting different levels of predictability (high vs. low) to mimic closely what occurs in natural languages and to get new insights into the changes that the mechanisms underlying auditory statistical learning (aSL) might undergo through the development. The participants performed the aSL task first under implicit and, subsequently, under explicit conditions to further analyze if children take advantage of previous knowledge of the to-be-learned regularities to enhance SL, as observed with the adult participants. These findings would also contribute to extend our knowledge of the mechanisms available to assist SL at each developmental stage. Although behavioral signs of learning, even under explicit conditions, were only observed for the adult participants, ERP data showed evidence of online segmentation in the brain in both groups, as indexed by modulations in the N100 and N400 components. A detailed analysis of the neural data suggests, however, that adults and children rely on different mechanisms to assist the extraction of word-like units from the continuous speech stream, hence supporting the view that SL with auditory linguistic materials changes through development.


Synthesis and DNase I inhibitory properties of new benzocyclobutane-2,5-diones.
Aim: Eight new benzocyclobutane-2,5-diones (1a-1h) were synthesized, and their inhibitory properties against bovine pancreatic DNase I were examined in vitro. Methods & results: Compounds 1a-1h were synthesized using photocycloaddition of duroquinone with various phenyl-substituted ethylenes in the presence of 18W compact fluorescent lamp (visible light). Two compounds, 1,3,4,6-tetramethyl-7-phenylbicyclo[4.2.0]oct-3-ene-2,5-dione (1a) and 1,3,4,6-tetramethyl-7-p-tolylbicyclo[4.2.0]oct-3-ene-2,5-dione (1b) inhibited DNase I in a noncompetitive manner with IC50 values below 150 μM and showed to be more potent DNase I inhibitors than crystal violet, used as a positive control. In order to analyze potential binding sites for the studied compounds with DNase I, molecular docking study was performed. Conclusion: The studied benzocyclobutane-2,5-diones offer a good starting point for a design of new DNase I inhibitors.


The preparation and characterisation of hetero- and homobimetallic complexes containing bridging naphthalene-1,8-dithiolato ligands.
Homo- and heterobimetallic complexes of the form [(PPh(3))(2)(mu(2)-1,8-S(2)-nap){ML(n)}] (in which (1,8-S(2)-nap)=naphtho-1,8-dithiolate and {ML(n)}={PtCl(2)} (1), {PtClMe} (2), {PtClPh} (3), {PtMe(2)} (4), {PtIMe(3)} (5) and {Mo(CO)(4)} (6)) were obtained by the addition of [PtCl(2)(NCPh)(2)], [PtClMe(cod)] (cod=1,5-cyclooctadiene), [PtClPh(cod)], [PtMe(2)(cod)], [{PtIMe(3)}(4)] and [Mo(CO)(4)(nbd)] (nbd=norbornadiene), respectively, to [Pt(PPh(3))(2)(1,8-S(2)-nap)]. Synthesis of cationic complexes was achieved by the addition of one or two equivalents of a halide abstractor, Ag[BF(4)] or Ag[ClO(4)], to [{Pt(mu-Cl)(mu-eta(2):eta(1)-C(3)H(5))}(4)], [{Pd(mu-Cl)(eta(3)-C(3)H(5))}(2)], [{IrCl(mu-Cl)(eta(5)-C(5)Me(5))}(2)] (in which C(5)Me(5)=Cp*=1,2,3,4,5-pentamethylcyclopentadienyl), [{RhCl(mu-Cl)(eta(5)-C(5)Me(5))}(2)], [PtCl(2)(PMe(2)Ph)(2)] and [{Rh(mu-Cl)(cod)}(2)] to give the appropriate coordinatively unsaturated species that, upon treatment with [(PPh(3))(2)Pt(1,8-S(2)-nap)], gave complexes of the form [(PPh(3))(2)(mu(2)-1,8-S(2)-nap){ML(n)}][X] (in which {ML(n)}[X]={Pt(eta(3)-C(3)H(5))}[ClO(4)] (7), {Pd(eta(3)-C(3)H(5))}[ClO(4)] (8), {IrCl(eta(5)-C(5)Me(5))}[ClO(4)] (9), {RhCl(eta(5)-C(5)Me(5))}[BF(4)] (10), {Pt(PMe(2)Ph)(2)}[ClO(4)](2) (11), {Rh(cod)}[ClO(4)] (12); the carbonyl complex {Rh(CO)(2)}[ClO(4)] (13) was formed by bubbling gaseous CO through a solution of 12. In all cases the naphtho-1,8-dithiolate ligand acts as a bridge between two metal centres to give a four-membered PtMS(2) ring (M=transition metal). All compounds were characterised spectroscopically. The X-ray structures of 5, 6, 7, 8, 10 and 12 reveal a binuclear PtMS(2) core with PtM distances ranging from 2.9630(8)-3.438(1) A for 8 and 5, respectively. The napS(2) mean plane is tilted with respect to the PtP(2)S(2) coordination plane, with dihedral angles in the range 49.7-76.1 degrees and the degree of tilting being related to the PtM distance and the coordination number of M. The sum of the Pt(1)coordination plane/napS(2) angle, a, and the Pt(1)coordination plane/M(2)coordination plane angle, b, a+b, is close to 120 degrees in nearly all cases. This suggests that electronic effects play a significant role in these binuclear systems.


Antimicrobial natural products: an update on future antibiotic drug candidates.
Over the last decade, it has become clear that antimicrobial drugs are losing their effectiveness due to the evolution of pathogen resistance. There is therefore a continuing need to search for new antibiotics, especially as new drugs only rarely reach the market. Natural products are both fundamental sources of new chemical diversity and integral components of today's pharmaceutical compendium, and the aim of this review is to explore and highlight the diverse natural products that have potential to lead to more effective and less toxic antimicrobial drugs. Although more than 300 natural metabolites with antimicrobial activity have been reported in the period 2000-2008, this review will describe only those with potentially useful antimicrobial activity, viz. with MICs in the range 0.02-10 microg mL(-1). A total of 145 compounds from 13 structural classes are discussed, and over 100 references are cited.


Bidirectional Communication between the Pontine Nucleus Incertus and the Medial Septum Is Carried Out by Electrophysiologically-Distinct Neuronal Populations.
Theta oscillations are key brain rhythm involved in memory formation, sensorimotor integration, and control of locomotion and behavioral states. Generation and spatiotemporal synchronization of theta oscillations rely on interactions between brain nuclei forming a large neural network, which includes pontine nucleus incertus (NI). Here we identified distinct populations of NI neurons, based on the relationship of their firing to hippocampal waves, with a special focus on theta oscillations, and the direction and type of interaction with the medial septum (MS) in male, urethane-anesthetized rats. By recording NI neuronal firing and hippocampal LFP, we described NI neurons that fire action potentials in a theta phase-independent or theta phase-locked and delta wave-independent or delta wave-locked manner. Among hippocampal activity-independent NI neurons, irregular, slow-firing, and regular, fast-firing cells were observed, while hippocampal oscillation-/wave-locked NI neurons were of a bursting or nonbursting type. By projection-specific optotagging, we revealed that only fast-firing theta phase-independent NI neurons innervate the MS, rarely receiving feedback information. In contrast, the majority of theta-bursting NI neurons were inhibited by MS stimulation, and this effect was mediated by direct GABAergic input. Described NI neuronal populations differ in reciprocal connections with the septohippocampal system, plausibly forming separate neuronal loops. Our results suggest that theta phase-independent NI neurons participate in theta rhythm generation through direct innervation of the MS, while theta-bursting NI neurons further transmit the rhythmic signal received from the MS to stabilize and/or strengthen rhythmic activity in other structures.SIGNIFICANCE STATEMENT The generation and spatiotemporal synchronization of theta oscillations rely on interactions between nuclei forming a large neural network, part of which is the pontine nucleus incertus (NI). Here we describe that within NI there are populations of neurons that can be distinguished based on the relationship of their firing to hippocampal theta oscillations and delta waves. We show that these neuronal populations largely do not have reciprocal connections with the septohippocampal system, but form separate neuronal loops. Our results suggest that medial septum (MS)-projecting, fast-firing, theta phase-independent NI neurons may participate in theta rhythm generation through direct innervation of the MS, while theta-bursting NI neurons may further transmit the rhythmic signal received from the MS to other structures.


Surgical microanatomy of the occipital artery for suboccipital muscle dissection and intracranial artery reconstruction.
The occipital artery (OA) is an important donor artery for posterior fossa revascularization. Harvesting the OA is difficult in comparison to the superficial temporal artery because the OA runs between suboccipital muscles. Anatomical knowledge of the suboccipital muscles and OA is essential for harvesting the OA during elevation of the splenius capitis muscle (SPL) for reconstruction of the posterior inferior cerebellar artery. We analyzed the running pattern of the OA and its anatomic variations using preoperative and intraoperative findings. From April 2012 to March 2018, we surgically treated 162 patients with suboccipital muscle dissection by OA dissection using the lateral suboccipital approach. The running pattern and relationship between the suboccipital muscles and OA were retrospectively analyzed using the operation video and preoperative enhanced computed tomography (CT) images. The anatomic variation in the running pattern of the OA was classified into two types: lateral type, running lateral to the muscle and medial type, running medial to the longissimus capitis muscle (LNG). The medial pattern was observed in 107 (66%) patients and the lateral pattern in 54 (33.3%); 1 (0.6%) patient had the OA running between the LNGs. Preoperative CT is effective in determining the running course of the OA, which is important for safely harvesting the OA during SPL elevation. There is a risk of causing OA injury in patients with the lateral pattern. This is the first report showing that the OA rarely runs in between the LNGs.


Relationships between changes in leptin and insulin resistance levels in obese individuals following weight loss.
Obesity can augment insulin resistance (IR), leading to increased risk of diabetes and heart disease. Leptin, ghrelin, and various fatty acids present in the cell membrane may modulate IR. In this study, we aimed to investigate the impact of weight loss on IR, serum leptin/ghrelin levels, and erythrocyte fatty acids, and studied the associations between changes in these variables. A total of 35 obese (body mass index ≥ 27) adults participated in a weight loss program for 3 months. IR was assessed using homeostasis model assessment for insulin resistance (HOMA-IR). The obese participants had a mean weight loss of 5.6 ± 3.8 kg followed by a 16.7% and 23.3% reduction in HOMA-IR and leptin (p < 0.001) levels, and an 11.3% increase in ghrelin levels (p = 0.005). The level of erythrocyte saturates decreased by 2.8%, while the level of n-3 polyunsaturates increased by 16.8% (all p < 0.05). The changes in leptin levels (-5.63 vs. -1.57 ng/mL) were significantly different (p = 0.004) in those with improved IR (changes in HOMA-IR < 0) than those without improvement (changes in HOMA-IR ≥ 0), though there were no differences in the changes of ghrelin (p = 0.120) and erythrocyte fatty acids (all p > 0.05) levels. After adjusting for age, gender, changes in ghrelin, and body fat, we found a significant correlation between decreases in leptin and less risk of no improvement in HOMA-IR levels [odds ratio (OR) = 0.69, p = 0.039]. In conclusion, a moderate weight reduction in obese participants over a short period significantly improved IR. This weight reduction concomitantly decreased serum leptin, increased ghrelin, and elevated some erythrocyte unsaturates. Only leptin correlated independently with IR improvement upon multivariable logistic regression analysis, which indicates that leptin may play a role in the modulation of IR following weight loss.


Influence of gender on diaphragm thickness using a method for determining intima media thickness in healthy young adults.
To clarify the differences in diaphragm thickness between male and female participants in healthy young adults with ultrasonography using the mean intima media thickness (IMT) method and to investigate the relationship between diaphragm thickness and respiratory pressure. Twenty-nine healthy individuals (16 females and 13 males) participated in the study. Diaphragm thickness was measured at total lung capacity (TLC) and at functional residual capacity (FRC) in each participant. We measured the diaphragm thickness using a method for mean intima media thickness. Moreover, change ratio of diaphragm thickness was calculated with the diaphragm thickness at TLC and FRC. Mean diaphragm thicknesses at FRC in males were significantly narrower than those in females (p < 0.001). The change ratio of diaphragm thickness was significantly augmented in males compared with that in females (p < 0.001). There was a positive correlation between the change ratio of diaphragm thickness and pulmonary function data and respiratory muscle strength in healthy young adults. The change ratio of diaphragm thickness using the IMT method can be accurately performed with a high degree of reproducibility by clinical laboratory technicians and may be a useful indicator for evaluating diaphragm muscle strength.


Three-Dimensional Hierarchical Seaweed-Derived Carbonaceous Network with Designed g-C3N4 Nanosheets: Preparation and Mechanism Insight for 4-Nitrophenol Photoreduction.
The development of g-C3N4-based photocatalysts with abundant active sites is of great significance for photocatalytic reactions. Herein, a smart and robust strategy was presented to fabricate three-dimensional (3D) g-C3N4 nanosheet-coated alginate-based hierarchical porous carbon (g-C3N4@HPC), including coating melamine on calcium alginate (CA) hydrogel beads, freeze-drying hydrogel beads as well as pyrolysis at high temperatures. The resulting photocatalyst possessed a significantly high surface area and a large amount of interconnected macropores compared with porous carbon without the melamine coating. The unique structural features could effectively inhibit the curling and agglomeration of g-C3N4 nanosheets, provide abundant photocatalytic active sites, and promote mass diffusion. Therefore, the g-C3N4@HPC composite exhibited remarkable photocatalytic activity and outstanding stability toward the photoreduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP) by NaBH4 under natural sunlight and simulated visible-light irradiation (λ > 420 nm) using a 300 W xenon lamp. Moreover, the mechanism toward the photocatalytic reaction was extensively studied by quenching experiments and electron spin resonance (ESR) experiments. The results showed that active hydrogen species were able to be achieved by following a dual-channel pathway in the NaBH4 system, which included photocatalytic reduction of H+ ions and photocatalytic oxidation of BH4- ions. This work not only opens up a new way to design efficient photocatalysts for various reactions but also provides a reference for an in-depth study of the photoreduction mechanism.


Gastric tonometry.
Gastric tonometry was originally proposed to assess the splanchnic perfusion. Several technological improvements have been introduced over the years and, to date, the preferred way to estimate the splanchnic perfusion is to rely on the arterial-gastric PCO2 gap. In this brief review we will discuss the value of the gastric tonometry, its physiological background and the clinical results observed so far.


[Photorefractive excimer laser keratectomy in myopia and astigmatism with a 2-year follow-up after surgery].
In a group of 1037 operated eyes the effectiveness, predictive value, stability and safety of photorefraction keratectomy by means of an excimer laser was tested in myopia (PRK) and astigmatism (PARK) with a two-year follow-up period after operation. The range of the preoperative refraction defect was -1.75 to -2.10 dpt (SE). The maximal value of the corrected cylinder in astigmatism was -6.0 cyl dpt. In the PRK group 459 operations were evaluated, in the PARK group 578 operations. Both groups are classified according to the spherical equivalent of the refractory defect into three groups. Group A is formed by myopia up to -6.0 dpt, group B myopia -6.25 to -10.0 dpt and group C above -10.25 dpt. Non-corrected visual acuity 6/12 and better was recorded in the 20th month after operation in group A in 93.4-97.6% (PARK-PRK) operated eyes, in group B 88.7-91.5% and in group C 61.2-68.4% of operated eyes. The achieved final refraction (SE) in an interval +(-1.0) dpt from the originally planned one was recorded during the 20th month after operation in group A in 92.8-96.5% (PARK-PRK) operated eyes, in group B 73.5-78.9% and in group C 59.7-63.5% operated eyes. The best corrected visual acuity (NKZO) in the whole group 20 months after operation was unaltered or better by 1 line of projected optotypes as compared with NKZO before operation in group A in 97.6%, in group B in 94.3% and in group C in 89.7% operated eyes. The analysis of results in PRK and PARK in the group with a two-year follow-up period after operation permits to evaluate the method as sufficiently effective and safe for correction of myopia up to -10.0 dpt but less predictive in myopia above -10.0 dpt.


All-Dielectric Metasurface Lenses for Achromatic Imaging Applications.
Metasurface can use artificial microstructures to manipulate electromagnetic waves more accurately and flexibly. All-dielectric metalens have a wide range of materials and low cost so it has a wide application prospect. Herein, we propose a all-dielectric achromatic metalens built with Si as the structural unit that can operate over a broadband of wavelengths in the visible region. It controls the wavefront of light through the Pancharatnam-Berry phase and propagation phase to eliminate the chromatic aberration. Meanwhile, we also use Gerchberg-Saxton algorithm and its improved algorithm to iterate over multiple design wavelengths and obtain holographic phases suitable for broadband. Thus, both the metalenses and holographic metasurfaces can achieve achromatic broadband in the visible light range, which provides a new method for the development of meta-optical imaging devices.


Basis Set Dependence of Optical Rotation Calculations with Different Choices of Gauge.
In this work, the basis set dependence of optical rotation (OR) calculations is examined for various choices of gauge/level of theory. The OR is calculated for a set of 50 molecules using B3LYP and CAM-B3LYP and 17 molecules using coupled cluster with single and double excitations (CCSD). The calculations employ the correlation-consistent basis sets, aug-cc-pVζZ with ζ = D, T, Q. An inverse-power extrapolation formula is then utilized to obtain OR values at the complete basis set (CBS) limit. We investigate the basis set convergence for these methods and three choices of gauge: length gauge (with gauge-including atomic orbitals, LG(GIAOs), for DFT), the origin-invariant length gauge [LG(OI)], and the modified velocity gauge (MVG). The results show that all methods converge smoothly to the CBS limit and that the LG(OI) approach has a slightly faster convergence rate than the other choices of gauge. While the DFT methods reach gauge invariance at the CBS limit, CCSD does not. The significant difference between the MVG and LG(OI) results at the CBS limit, 26%, indicates that CCSD is not quite at convergence in the description of electron correlation for this property. On the other hand, gauge invariance at the CBS limit for DFT does not lead to the same OR values for the two density functionals, which is also due to electron correlation incompleteness. A limited comparison to gas-phase experimental OR values for the DFT methods shows that CAM-B3LYP seems more accurate than B3LYP. Overall, this study shows that the LG(OI) approach with the aug-cc-pVTZ basis set for DFT, and with the CBS(DT) extrapolation for CCSD, provides a good cost/accuracy balance.


Fast identification of thermostable beta-glucosidase mutants on cellobiose by a novel combinatorial selection/screening approach.
Engineering costly cellulases on natural cellulosic substrates is of importance for emerging biomass-based biorefineries. Directed enzyme evolution is becoming a popular tool, but identification of desired mutants from a large mutant library remains challenging sometimes. In this work, we demonstrated a novel combinatorial selection/screening strategy for finding thermostable beta-glucosidase on its natural substrate-cellobiose. First, selection was conducted through complementation of beta-glucosidase for non-cellobiose-utilizing Escherichia coli so that only the cells expressing active beta-glucosidase can grow on a M9 synthetic medium with cellobiose as the sole carbon source (selection plate). Second, the clones on the selection plates were duplicated by using nylon membranes. After heat treatment, the nylon membranes were overlaid on M9/cellobiose screening plates so that remaining activities of thermostable beta-glucosidase mutants hydrolyzed cellobiose on the screening plates to glucose. Third, the growth of an indicator E. coli strain that can utilize glucose but not cellobiose on the screening plates helped detect the thermostable beta-glucosidase mutants on the selection plates. Several thermostable mutants were identified from a random mutant library of the Paenibacillus polymyxa beta-glucosidase. The most thermostable mutant A17S had an 11-fold increase in the half-life of thermoinactivation at 50 degrees C.


Synthesis of a functional F0 sector of the Escherichia coli H+-ATPase does not require synthesis of the alpha or beta subunits of F1.
The uncB, E, F, and H genes of the Escherichia coli unc operon were cloned behind the lac promoter of plasmid pUC9, generating plasmid pBP101. These unc loci code, respectively, for the chi, omega, and psi subunits of the F0 sector and the delta subunit of the F1 sector of the H+-ATP synthase complex. Induction of expression of the four unc genes by the addition of isopropyl-beta-D-thiogalactoside resulted in inhibition of growth. During isopropyl-beta-D-thiogalactoside induction, the three subunits of F0 were integrated into the cytoplasmic membrane with a resultant increase in H+ permeability. A functional F0 was formed from plasmid pBP101 in a genetic background lacking all eight of the unc structural genes coding the F1F0 complex. In the unc deletion background, a reasonable correlation was observed between the amount of F0 incorporated into the membrane and the function measured, i.e., high-affinity binding of F1 and rate of F0-mediated H+ translocation. This correlation indicates that most or all of the F0 assembled in the membrane is active. Although the F0 assembled under these conditions binds F1, only partial restoration of NADH-dependent or ATP-dependent quenching of quinacrine fluorescence was observed with these membranes. Proteolysis of a fraction of the psi subunit may account for this partial deficiency. The experiments described demonstrate that a functional F0 can be assembled in vivo in E. coli strains lacking genes for the alpha, beta, gamma, and epsilon subunits of F1.


Japanese encephalitis virus capsid protein interacts with non-lipidated MAP1LC3 on replication membranes and lipid droplets.
Microtubule-associated protein 1 light chain 3 (MAP1LC3) is a protein with a well-defined function in autophagy, but still incompletely understood roles in several other autophagy-independent processess. Studies have shown MAP1LC3 is a host-dependency factor for the replication of several viruses. Japanese encephalitis virus (JEV), a neurotropic flavivirus, replicates on ER-derived membranes that are marked by autophagosome-negative non-lipidated MAP1LC3 (LC3-I). Depletion of LC3 exerts a profound inhibition on virus replication and egress. Here, we further characterize the role of LC3 in JEV replication, and through immunofluorescence and immunoprecipitation show that LC3-I interacts with the virus capsid protein in infected cells. This association was observed on capsid localized to both the replication complex and lipid droplets (LDs). JEV infection decreased the number of LDs per cell indicating a link between lipid metabolism and virus replication. This capsid-LC3 interaction was independent of the autophagy adaptor protein p62/Sequestosome 1 (SQSTM1). Further, no association of capsid was seen with the Gamma-aminobutyric acid receptor-associated protein family, suggesting that this interaction was specific for LC3. High-resolution protein-protein docking studies identified a putative LC3-interacting region in capsid, 56FTAL59, and other key residues that could mediate a direct interaction between the two proteins.


Clinical response to dabrafenib plus trametinib in a pediatric ganglioglioma with BRAF p.T599dup mutation.
In this follow-up report, we present updated information regarding a previously reported pediatric patient with a World Health Organization grade I ganglioglioma harboring a BRAF p.T599dup mutation (Cold Spring Harb Mol Case Stud 4: a002618). This patient, based on our initial finding, is receiving combination targeted therapy with a selective BRAF inhibitor (dabrafenib) plus MEK inhibitor (trametinib). The combination therapy was started after the patient experienced progressive tumor growth and worsening neurological symptoms, including visual changes, headaches, and peripheral neuropathy, despite 9 months of treatment with adjuvant chemotherapy (vinblastine). The patient has been receiving dabrafenib plus trametinib for 15 months and continues to have stable disease as well as improved neurological symptoms. Although combinatorial therapy targeting BRAF and MEK using dabrafenib and trametinib, respectively, is indicated for tumors harboring a BRAF p.V600E/K mutation, our report demonstrates efficacy of this combination in a non-V600E BRAF-mutated tumor. The identification of BRAF alterations may assist clinicians in determining alternative targeted treatment strategies, especially considering the paucity of effective treatments for primary brain tumors and the poor prognosis associated with many central nervous system (CNS) diagnoses. Additional case studies or larger cohort reports will continue to clarify the efficacy of BRAF and/or MEK inhibitors in patients whose tumors harbor a BRAF alteration.


Compound I reactivity defines alkene oxidation selectivity in cytochrome P450cam.
Prediction of the chemoselectivity of drug oxidation by the human cytochrome P450 enzymes will aid in the avoidance of adverse drug reactions. The chemoselectivity of alkene oxidation is an important problem to address, as it can result in the formation of epoxides, which can have toxic effects. In this paper the epoxidation and hydroxylation of cyclohexene and propene by the bacterial P450(cam) isoform are modeled with hybrid quantum mechanical/molecular mechanical (QM/MM) methods. Snapshots for QM/MM modeling are chosen from molecular dynamics trajectories, to sample the different conformations of the enzyme-substrate complex. The energy barriers obtained for these processes are in qualitative agreement with experimental work, supporting the use of QM/MM methods in the study of selectivity for this class of enzyme. This work highlights the complexity involved in modeling these systems with QM/MM and the importance in the selection of starting geometries.


Food-Based Dietary Guidelines and Protein Quality Definitions-Time to Move Forward and Encompass Mycoprotein?
Food-Based Dietary Guidelines (FBDG) lack uniformity globally, with the integration of protein food sources being highly variable. Protein guidance tends to be dichotomous, e.g., animal versus plant with other categories such as fungal proteins being overlooked. In 2019 the EAT Lancet Food in the Anthropocene report was a chief driver questioning the need to supply healthy diets from sustainable food systems. Some countries are developing FBDG that integrate these aspects, but these are quite often protracted, too subtle or misaligned with other countries, diluting the effects of meaningful global change. Protein quality metrics also underpin the dissemination of dietary guidance. However, for protein, these remain based on a food's essential amino acid profile and digestibility scores, thus are nutritionally and physiologically centric. It has been proposed that this definition is becoming increasingly myopic from a wider societal perspective. Updated indices should include contemporary issues such as protein diversity and environmental outcomes. Taken together, there is opportunity for renewed thinking about both FBDG and protein quality definitions, with scope to include both health and environmental outcomes and need to move towards the concept of protein diversification.


A cholestatic diagnostic dilemma.
A 28-year-old man with hereditary spherocytosis presented with abdominal pain and jaundice. He had severe, mainly conjugated (642 μmol/l), hyperbilirubinaemia (1033 μmol/l), with elevated liver enzymes: alkaline phosphatase (ALP) (163 IU/l), γ-glutamyltransferase (gGT) (277 IU/l) and aspartate transaminase (AST) (358 IU/l). Abdominal ultrasound and magnetic resonance cholangiopancreatography (MRCP) showed gallstones in the gallbladder but an absence of biliary duct dilation. Liver biopsy was consistent with cholestasis but showed no large duct obstruction. The cause of the cholestasis was unclear, was it a primary intrahepatic pathology or secondary to a posthepatic cause? He presented with similar symptoms days later with gallstones in the bile duct (choledocholithiasis) and underwent endoscopic retrograde cholangiopancreatography (ERCP) and cholecystectomy. This report guides one through the assessment of jaundice and serves as an example whereby the diagnosis of a common cause of illness is blurred by an atypical clinical presentation and relevant comorbidities. A diagnosis of benign recurrent intrahepatic cholestasis (BRIC) is also considered.


[A Case of Rocuronium-induced Anaphylaxis in Which Surgery was Subsequently Performed under General Anesthesia without Neuromuscular Blocking Agents].
We report here a case of rocuronium-induced anaphylactic shock in a 41-year-old woman. She was scheduled for partial hepatectomy due to liver metastasis of a pheochromocytoma. Anesthesia was induced with propofol, remifentanil, and rocuronium. Bag-mask ventilation was difficult, and her blood pressure fell to around 40 mmHg just after induction. Subsequently, her trachea was intubated and adrenaline was injected. However, due to the subsequent persistence of severe hypotension and hypoxia, cardiopulmonary resuscitation was necessary. Suspecting the development of pulmonary embolism or anaphylaxis, we performed transesophageal echography; however, no evidence of right heart dilatation was observed, indicating a low possibility of pulmonary embolism. Although her general condition was stabilized, surgery was canceled. Blood tests showed high serum histamine and tryptase levels, suggesting an Ig-E mediated allergic reaction. A skin test performed five weeks after anesthesia suggested that she had suffered from rocuronium-induced anaphylaxis. A skin test also showed cross-reactivity between rocuronium and vecuronium. Therefore, we did not use any neuromuscular agent for the subsequent surgery, which was completed uneventfully. Determining the drug responsible for anaphylaxis helps to prevent recurrence of anaphylaxis.


Long-term changes in cyclosporine pharmacokinetics after renal transplantation in children: evidence for saturable presystemic metabolism and effect of NR1I2 polymorphism.
To improve cyclosporine dose individualization, the authors carried out a comprehensive analysis of the effects of clinical and genetic factors on cyclosporine pharmacokinetics in 176 children before and up to 16 years after renal transplantation. Pretransplantation test doses of cyclosporine were given intravenously and orally, followed by blood sampling for 24 hours. After transplantation, cyclosporine was quantified at trough, 2 hours postdose, or with dose-interval curves. A 3-compartment population pharmacokinetic model was used to describe the data. Cyclosporine oral bioavailability increased more than 1.5-fold in the first month after transplantation, returning thereafter gradually to its initial value in 1 to 1.5 years. Moreover, older children receiving cyclosporine twice daily as the gelatin capsule microemulsion formulation had an about 1.25 to 1.3 times higher bioavailability than did the younger children receiving the liquid formulation thrice daily. In 91 children with genetic data after transplantation, patients carrying the NR1I2 g.-25385C-g.-24381A-g.-205_-200GAGAAG-g.7635G-g.8055C haplotype had about one-tenth lower bioavailability, per allele, than did noncarriers (P = .039). The significance of the NR1I2 genotype warrants further study. In conclusion, by accounting for the effects of developmental factors (body weight), time after transplantation, and cyclosporine dosing frequency/formulation, it may be possible to improve individualization of cyclosporine dosing in children.


Chemical influences on adsorption-mediated self-propelled drop movement.
We report studies of reactive wetting employing droplets of a nonpolar liquid (decahydronaphthalene) on chemically patterned surfaces. The drops contain an n alkylamine that adsorbs onto surfaces exposing carboxylic acid groups and produces surfaces exposing methyl groups. The change in surface energy that occurs concurrent with the formation of an oriented monomolecular film of alkylamine during this process is sufficient to produce a self-propelled movement of decahydronaphthalene drops on the surface. We employed patterning to direct the movement of the drops on the surface, thereby allowing measurements of the relationships between the macroscopic fluidic behavior of the droplets and microscopic adsorption events. Specifically, we examined the effects of the unbalanced surface-tension force and the influences of adsorbate concentration on drop movement. In this latter case, both kinetic and thermodynamic arguments can be applied to describe the system. We compared the predictions from these two approaches by analyzing data from the present system and those reported by F. Domingues Dos Santos and T. Ondarcuhu [Phys. Rev. Lett. 75(16), 2972 (1975)] that exhibited opposite trends in behavior. The present analysis provides insight into the influence of chemical reaction kinetics on adsorption-mediated drop movement (i.e., reactive wetting).


Effect of curvature on C-F bonding in fluorinated carbons: from fullerene and derivatives to graphite.
The effect of the curvature of the carbon lattice is discussed taking into account NMR data on various fluorinated carbons including C(60) fullerenes, single, double and multiwall carbon nanotubes. Graphite fluorides and highly fluorinated fullerenes are used as limit model compounds for planar and spherical geometries, respectively. The curvature results in a weakening of the C-F bonding covalence. First of all, various highly fluorinated fullerenes with increasing F/C molar ratio were prepared by treatment with pure gaseous fluorine. A preliminary study using XRD, EPR and IR spectroscopy confirms that the highest fluorination level can be reached either at 133 or at 300 degrees C. In order to extract the correlation between fluorine and carbon atoms and the C-F bond length, specific sequences such as solid echo, two-dimensional (19)F -->(13)C cross polarization wide-line separation and inverse (19)F -->(13)C cross polarization were also used for fluorinated C(60).


Experimental susceptibility of Atlantic cod, Gadus morhua (L.), and Atlantic halibut, Hippoglossus hippoglossus (L.), to different genotypes of viral haemorrhagic septicaemia virus.
Viral haemorrhagic septicaemia (VHS) is a well-characterized disease of rainbow trout, Oncorhynchus mykiss, which has also caused economic losses in marine turbot farms in the British Isles. We have previously demonstrated that turbot, Scophthalmus maximus, are susceptible to isolates of viral haemorrhagic septicaemia virus (VHSV) that are endemic in the marine environment, highlighting a potential risk to marine aquaculture. Given the increasing interest in the intensive rearing of additional aquaculture species such as Atlantic cod, Gadus morhua, and Atlantic halibut, Hippoglossus hippoglossus, this study aimed at investigating the susceptibility of these species to VHSV. Both species were found to be largely resistant to VHS following immersion challenge with a selection of 18 isolates, representing the known marine VHSV genotypes. Only one and two VHSV-associated mortalities occurred out of a total of 1710 and 1254 halibut and cod, respectively. These findings suggest that there is a low direct risk to the development of commercial cod and halibut aquaculture from the existing endemic reservoir of VHSV. This study, coupled to field observations has, however, highlighted the fact that both species can become infected with VHSV. The known adaptability of RNA viruses, together with the selection pressures associated with intensive aquaculture would thus advocate a cautious approach to VHSV surveillance within these emerging industries.


Sorption of lead and thallium on borosilicate glass and polypropylene: Implications for analytical chemistry and soil science.
Adsorption of 10-100 ppb lead and thallium by borosilicate glass and polypropylene surfaces was studied. No thallium was adsorbed by either substrate at pH 7.0. About 50-60% of the lead was adsorbed by borosilicate glass at pH 5.5 and pH 7.0, with statistically indistinguishable conditional equilibrium constants (KD* values) of 1.47±0.40 mL cm-2 and 1.80±0.11 mL cm-2, respectively. Polypropylene adsorbed significantly more lead than borosilicate glass at pH 7.0 (80.9%, KD* =5.33±0.45 mL cm-2), and substantially less at pH 5.5 (16.7%, KD* = 0.27±0.03 mL cm-2), indicating a fundamental change in the nature of the polypropylene surface. All adsorption data fitted Freundlich plots. Preliminary experiments indicate that at pH 5.5, a fraction of the lead sequestered by the borosilicate glass may be bound irreversibly by migrating into the glass matrix. The results suggest that tectosilicates and silicate glasses may represent important adsorbents of some heavy metals in soils, despite showing very low intrinsic cation exchange capacities.


Novel treatments under development for muscular dystrophy.
Extract: There is no specific therapy for the muscular dystrophies, a group of genetic diseases that affect primarily skeletal muscle and compromise patients' quality of life and, in the most severe forms, their survival. New results coming from gene and cell therapy, as well as novel strategies aimed at preventing or delaying muscle degeneration, have raised hopes that clinical experimentation may start in the near future. Muscular dystrophies (MD) primarily affect the skeletal muscle. They are clinically and molecularly diverse and, in the most severe forms, such as Duchenne Muscular Dystrophy (DMD), lead the patient to wheelchair dependency, respiratory failure and premature death. In many cases, the mutation impairs genes that encode proteins forming a complex that links the cytoskeleton to the basal membrane, a collagen scaffold in which cells are embedded. Mutation in one of these proteins often causes a breakdown of the whole complex, leading to an increased fragility of the membrane, especially during intense contractile activity. This, in turn, results in increased calcium entry and subsequently focal or diffuse damage to the muscle fiber.


Correlation between low Gleason score and prostate specific antigen levels with incidence of bone metastases in prostate cancer patients: when to omit bone scans?
To identify correlation and incidence of bone metastases in prostate cancer patient with low Gleason scores (GS) and prostate specific antigen (PSA) levels. This descriptive restrospective study covered patients with prostate cancer in Cipto Mangunkusumo Hospital in 2006-2011. Of a total of 478, those who had PSA values, histological examination, and bone scan were included, resulting in 358 eligible cases. PSA values were measured using the sandwich electrochemiluminescent immunoassay. Histological examination was graded according to Gleason's grading system and divided into 3 categories: well differentiated (GS ≤ 6), moderately differentiated (GS 7) and poorly differentiated (GS 8-10). Bone scans were performed using a radiopharmaceutical agent (Tc 99m methylenen diphosphonate) with images captured by gamma camera. The mean age was 67.5 ± 7.8, mean GS was 7.7 ± 1.3 and median PSA was 56.9 (range: 0.48-17000 ng/ mL). There were 11 patients (3.0%) with positive bone scan with PSA<20 ng/mL and GS<8. Furthermore, there were 2 patients (0.6%) with GS ≤ 6 and PSA<10 ng/mL showing bone metastasis. In our study, there were still small percentage of patients with bone metastasis even when low values of PSA (PSA<10 ng/mL) and GS (GS ≤ 6) were applied.


New horizons in the perioperative care of older adults.
Older adults undergoing surgery have high perioperative morbidity and mortality. Age-related physiological changes and prevalence of geriatric syndromes such as frailty increase the risk of adverse postoperative outcomes. Geriatricians utilise comprehensive geriatric assessment (CGA) and management to identify and manage geriatric syndromes, and deliver patient-centred perioperative care. Perioperative models of CGA are established for older patients undergoing hip fracture surgery. Recent trials support the benefits of perioperative models of CGA for non-orthopaedic surgery, and have influenced current care recommendations for older surgical patients. Areas for further action include addressing the implementation gap between recommended evidence-based perioperative care and routine perioperative care, evaluating the clinical and cost-effectiveness of perioperative models of CGA for patients living with frailty, and embedding routine use of patient-reported outcome measures to inform quality improvement.


Treatment of Venous Thromboembolism With New Anticoagulant Agents.
Venous thromboembolism (VTE) is a common disease associated with high risk for recurrences, death, and late sequelae, accounting for substantial health care costs. Anticoagulant agents are the mainstay of treatment for deep vein thrombosis and pulmonary embolism. The recent availability of oral anticoagulant agents that can be administered in fixed doses, without laboratory monitoring and dose adjustment, is a landmark change in the treatment of VTE. In Phase III trials, rivaroxaban, apixaban, edoxaban (antifactor Xa agents), and dabigatran (an antithrombin agent) were noninferior and probably safer than conventional anticoagulation therapy (low-molecular-weight heparin followed by vitamin K antagonists). These favorable results were confirmed in specific patient subgroups, such as the elderly and fragile. However, some patients, such as those with cancer or with intermediate- to high-risk pulmonary embolism, were underrepresented in the Phase III trials. Further clinical research is required before new oral anticoagulant agents can be considered standard of care for the full spectrum of patients with VTE.


Down-regulation of CD40 and CD80 on B cells in patients with life-threatening systemic lupus erythematosus after successful treatment with rituximab.
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoreactive T cells and polyclonally activated B cells that produce autoantibodies. Five SLE patients who failed to respond to conventional immunosuppressants were treated with anti-CD20 antibody (rituximab) and their clinical manifestations and laboratory data were evaluated, including phenotypic analysis of B cells. Rituximab (375 mg/m(2)) was administered weekly for 2 weeks in five SLE patients who developed severe manifestations despite intensive treatment. Rituximab resulted in rapid improvement (within several days) in clinical manifestations such as consciousness disorder, seizures, progressive sensory disorder, haemolytic crisis, cardiac function and laboratory data. The effects lasted 20 months in one patient; other patients were in remission for more than 6 months. Flow cytometric analysis revealed down-regulation of CD40 and CD80 expression on CD19-positive B cells 1 week after infusion of rituximab, and such down-regulation was seen for more than 7 months in two patients. Our pilot study provides sufficient evidence of excellent tolerability and high efficacy of rituximab therapy in refractory SLE. Rituximab not only reduced B-cell number and IgG levels but down-regulated CD40 and CD80 on B cells, suggesting possible disturbance of T-cell activation through these costimulatory molecules. Reduction of both quantity and quality of B cells suggests that rituximab could improve the disease course in patients with refractory SLE.


Migration--a challenge to modern citizenship.
"I will treat the question of whether there is an inherent difficulty hindering the nation-state and its concept of citizenship from coping with the problem of migration.... I interpret democracy as a political form which bridges the gap between the premodern concept of republicanism, which inspires the idea of citizenship, and statehood (which tends to be hostile to the idea of citizenship).... I claim also that democracy is necessarily harmonious with citizenship...which then leads me to the consequences of this judgment for the problem of migration...."


Do environmental and hereditary factors affect the psychophysiology and left-right shift in left-handers?
The principal objective of this study was to investigate the reality of sinistrality in left-handed subjects. The subjects were assessed by a 20-item questionnaire with two groups taken from Oldfield's and Yetkin questionnaires. The relation of different effects on left-hand preference was studied in men and women considering familial information and writing hand. The degree of hand preference was determined by Geshwind scores (GSs). The GS degrees of hand preferences were divided into weak and strong left-hand ranging from -20% to -100%. After assessing the GSs we asked the subjects to answer 22 questions with different aims, which were especially written for only left-handers. Some of them were based on familial sinistrality and hereditary relation to left-handers. The others concerned hand, foot, eye, shoulder, and ear preferences; the psychology of left-handedness; the knowledge concerned with left-handedness; the success, ability, and interest education and skill, and morphological differences in hand and foot sizes. In the present study, it was found that the subjects have had at least one left-handed person in their family or among their relatives. Some subjects had shifted their left hand preference in favor of their right hands. Nevertheless, the left-handers have been found as a presence with their own peculiarities.


Evaluation of the Long-Term Storage Stability of the Cyanide Antidote: Dimethyl Trisulfide and Degradation Product Identification.
This study reports the long-term storage stability of a formulation of the cyanide (CN) antidote dimethyl trisulfide (DMTS). The F3-formulated DMTS was stored in glass ampules at 4, 22, and 37 °C. Over a period of one year, nine ampules (n = 3 at each temperature) were analyzed by high-performance liquid chromatography (HPLC)-UV/vis at daily time intervals in the first week, weekly time intervals in the first month, and monthly thereafter for a period of one year to determine the DMTS content. No measurable loss of DMTS was found at 4 and 22 °C, and good stability was noted up to five months for samples stored at 37 °C. At 37 °C, a 10% (M/M) decrease of DMTS was discovered at the sixth month and only 30% (M/M) of DMTS remained by the end of the study; discoloration of the formulation and the growth of new peaks in the HPLC chromatogram were also observed. To identify the unknown peaks at 37 °C, controlled oxidation studies were performed on DMTS using two strong oxidizing agents: meta-chloroperoxybenzoic acid (mCPBA) and hydrogen peroxide (H2O2). Dimethyl tetrasulfide and dimethyl pentasulfide were observed as products using both of the oxidizing agents. Dimethyl disulfide was also observed as a product of degradation, which was further oxidized to S-methyl methanethiosulfonate only when mCPBA was used. HPLC-UV/vis and gas chromatography-mass spectrometry/solid phase microextraction analysis revealed good agreement between the degradation products of the stability study at 37 °C and those of disproportionation reactions. Furthermore, at 4 and 22 °C, chromatograms were remarkably stable over the one-year study period, indicating that the F3-formulated DMTS shows excellent long-term storage stability at T ≤ 22 °C.


Characterization by two-endpoint comparisons of the genetic toxicity profiles of vinyl chloride and related etheno-adduct forming carcinogens in Drosophila.
The genetic toxicity profiles of vinyl chloride (VCl), vinyl bromide (VBr), ethyl carbamate (EC), vinyl carbamate (VC) and some structurally related chemicals were investigated in both somatic and germ cells of Drosophila melanogaster. In the white/white+ eye mosaic assay, a screening system measuring predominantly homologous recombination in somatic cells, only marginal genotoxic activities were observed for acetyl chloride (ACl), glycolaldehyde (GCA), 2,2'-dichlorodiethyl ether (DDE) and methyl carbamate (MC), whereas VCl, 2-chloroacetaldehyde (CAA), VBr, 2-bromoacetaldehyde (BAA) and EC were clearly recombinogenic in the assay. Those chemicals proven to be recombinogenic in somatic cells were investigated further in postmeiotic male germ cells, utilizing as descriptors of their genotoxicity I(CL/RL) and M(exr-)/M(exr+) indices. The I(CL/RL) index is the rate of induced chromosome loss (CL), a clastogenic event, divided by the forward mutation rate, measured as recessive lethal (RL) mutations in 700 loci of the X-chromosome. The M(exr-)/M(exr+) mutation enhancement ratio is obtained by determining RL under excision repair deficient versus repair proficient conditions. With I(CL/RL) values (2.7-6.9) similar to those obtained for cross-linking agents, vinyl chloride, vinyl bromide, ethyl carbamate and vinyl carbamate are all efficient clastogenic agents in Drosophila germ cells. In the absence of excision repair, however, neither CEO nor CAA gave a hypermutability response (M(exr-)/M(exr+) approximately 1). By contrast, VCl, VBr, EC and VC showed clearly enhanced M(exr-)/M(exr+) ratios, suggesting that these compounds produce some repairable DNA modification(s) that are not generated by their epoxides. This unexpected finding points to the formation of other, yet unknown, metabolites of vinyl chloride, vinyl bromide, ethyl carbamate and vinyl carbamate. Our results support the concept that the epoxides chloroethylene oxide (CEO), bromoethylene oxide (BEO) and vinyl carbamate epoxide (VCO) are the most essential mutagenic intermediates. Compared to chloroethylene oxide (CEO), 2-chloroacetaldehyde (CAA) was approximately 50 times less effective in the induction of RL, whereas BAA was inactive as a mutagen. These findings are consistent with the general view that CAA and BAA play no major role in the genotoxic action of vinyl halides.


Is participation in labour market programmes related to mental health? Results from a 14-year follow-up of the Northern Swedish Cohort.
There is a lack of empirical studies assessing the possible impact of active labour market programmes (ALMP) on health. The aim of this study was to analyze whether participation in ALMP, in contrast to being unemployed and not participating in ALMP (UNALMP), was related to mental health at different ages. The study was carried out in a medium-sized industrial town in the north of Sweden. The cohort, consisting of all 1,083 pupils who attended or should have attended the last year of compulsory school in 1981, was followed up at the ages of 16, 18, 21 and 30. Data on 381 individuals at age 21, and 281 at age 30 were used in the study. The main health measurement was psychological symptoms among participants of ALMP in contrast to UNALMP at ages 21 and 30, and was analyzed by propensity score matching method (PSM) and multivariate logistic regression. Generally, ALMP had higher scores of psychological symptoms than UNALMP. Nevertheless, participation in ALMP was not related to mental health. Due to methodological shortages our results have to be interpreted with caution. Adjustment for either all background selection variables or the propensity score in multivariate logistic regression showed similar associations, suggesting that propensity score could be used to adjust for background selection variables. There is a need for more well-designed studies, using a theoretical framework, within the field, that are based on larger samples.


Multisite phosphorylation of ornithine decarboxylase in transformed macrophages results in increased intracellular enzyme stability and catalytic efficiency.
Ornithine decarboxylase (ODC) is the initial inducible enzyme in the polyamine biosynthetic pathway. In the transformed macrophage-derived RAW264 cell line, ODC was overproduced and existed in both unphosphorylated and phosphorylated forms. To date, the only protein kinase known to phosphorylate mammalian ODC is casein kinase II (CKII). ODC was phosphorylated in vitro by CKII and subjected to exhaustive sequential proteolysis with trypsin and V8 protease. Two-dimensional peptide mapping showed only a single phosphopeptide; two-dimensional phosphoamino acid analysis of the phosphopeptide revealed only 32P-labeled serine. ODC was metabolically radiolabeled with 32Pi in RAW264 cells and also subjected to proteolysis, two-dimensional peptide mapping, and phosphoamino acid analysis. Two phosphopeptides were generated from the metabolically radiolabeled ODC, including one that migrated similarly to the peptide phosphorylated by CKII in vitro. Each of the in situ radiolabeled ODC peptides contained both 32P-labeled serine and threonine residues. Thus, in RAW264 cells, ODC is phosphorylated on at least one serine residue in addition to that phosphorylated by CKII and on at least two threonine residues. Phosphorylated ODC had an increased stability to intracellular proteolysis compared with unphosphorylated ODC, their half-lives being 49.2 +/- 3.78 and 23.9 +/- 2.6 min (p = 0.001), respectively. The phosphorylated and unphosphorylated forms of ODC were independently purified to homogeneity. Kinetic analysis revealed that the catalytic efficiency of the phosphorylated form of ODC was 50% greater than that of the unphosphorylated form; the unphosphorylated ODC had a Vmax of 20.54 +/- 1.65 micromol/min/mg, whereas the phosphorylated form had a Vmax of 30.61 +/- 2.6 micromol/min/mg (p = 0.005). Phosphorylation of ODC by CKII has no effect on enzyme activity. Taken together, these findings demonstrate that regulation of ODC activity is governed by as yet unidentified protein kinases.


[Reduction of the amputation rate in bone and soft tissue sarcoma by interdisciplinary cooperation].
On the basis of three patients with bone and soft tissue sarcoma,we would like to illustrate the necessity of a well working inter-disciplinary cooperation of radiologists, internal oncologists, radiation therapists, orthopaedic surgeons and plastic surgeons. Functional extremity preservation in sarcoma patients can be achieved by a good interdisciplinary management without im-pairing the total prognosis of the patients. Patients with sarcoma should be treated in centres in which all specialised divisions are experienced and well trained in the treatment of tumour patients. Only then can a promising approach be achieved.


Human factors in crashes of commuter airplanes.
Pilots and frequent travelers on commuter aircraft are exposed to higher risks of death or injury than those on major air carriers. To provide a better understanding of the circumstances of crashes of scheduled commuter airplanes, National Transportation Safety Board data were analyzed for all cases of death, serious injury, or major damage involving commuter airplanes during 1983-88, when 172 people were killed and 207 injured in 118 events. Three-fourths of cases involved inadequate pilot performance, notably poor handling of emergencies and improper instrument flying procedures. Pilot errors occurred disproportionately in bad weather, which played a role in 30% of crashes. Aircraft malfunctions were involved in 42% of crashes. Certain airplanes were overinvolved in gear-up landings or in crashes due to fuel mismanagement. Greater priority should be given to applying known preventive measures to the problem of commuter crashes.


Protease-activated receptor 2 in dorsal root ganglion contributes to peripheral sensitization of bone cancer pain.
Treating bone cancer pain continues to be a major clinical challenge, and the underlying mechanisms of bone cancer pain remain elusive. Protease-activated receptor 2 (PAR2) has been reported to be involved in neurogenic inflammation, nociceptive pain and hyperalgesia. Here, we investigated the role of PAR2 in bone cancer pain development. Expression of PAR2, mechanical allodynia, thermal hyperalgesia and neurochemical alterations induced by bone cancer pain were analysed in male, adult C3H/HeJ mice with tumour cell implantation (TCI). To investigate the contribution of PAR2 to bone cancer pain, PAR2 antagonist peptide and PAR2 knockout mice were used. TCI produced bone cancer-related pain behaviours. Production and persistence of these pain behaviours were well correlated with TCI-induced up-regulation of PAR2 in sciatic nerve and dorsal root ganglia (DRG). PAR2 knockout and spinal administration of PAR2 antagonist peptide prevented and/or reversed bone cancer-related pain behaviours and associated neurochemical changes in DRG and dorsal horn (DH). TCI also induced proteases release in tumour-bearing tibia, sciatic nerve and DRG. Plantar injection of supernatant from sarcoma cells induced PAR2 up-regulation and intracellular calcium [Ca(2+) ]i increase in DRG, and calcitonin gene-related peptide accumulation in DH, as well as significant thermal and mechanical hyperalgesia, which were all in PAR2-dependent manners. These findings suggest that PAR2 may be a key mediator for peripheral sensitization of bone cancer pain. Inhibiting PAR2 activation, especially during the early phase, may be a new therapy for preventing/suppressing development of bone cancer pain.


Enhancement in T1 lipase purification recovery using the novel construct pGEX4T1/His-T1.
The Geobacillus zalihae strain T1 produces a thermostable T1 lipase that could be used for industrial purposes. Previously, the GST-T1 lipase was purified through two chromatographic steps: affinity and ion exchange (IEX) but the recovery yield was only 33%. To improve the recovery yield to over 80%, the GST tag from the pGEX system was replaced with a poly-histidine at the N-terminal of the T1 lipase sequence. The novel construct of pGEX/His-T1 lipase was developed by site-directed mutagenesis, where the XbaI restriction site was introduced upstream of the GST tag, allowing the removal of tag via double digestion using XbaI and EcoRI (existing cutting site in the pGEX system). Fragment of 6 × His-T1 lipase fusion was synthesized, cloned into the pGEX4T1 system, and expressed in Escherichia coli BL21 (DE3) pLysS, resulting in lipase-specific activity at 236 U/mg. The single purification step of His-T1 lipase was successfully achieved using nickel Sepharose 6FF with an optimized concentration of 5 mM imidazole for binding, yielding the recovery of 98%, 1,353 U/mg lipase activity, and a 5.7-fold increase in purification fold. His-T1 lipase was characterized and was found to be stable at pH 5-9, active at 70 °C, and optimal at pH 9.


[Cutaneous Herpesvirus Infection].
Among human herpes viruses, those known to cause neurological symptoms are the herpes simplex virus (HSV) types 1 and 2 and the varicella-zoster virus (VZV), which remain latent in the dorsal root ganglion. HSV causes herpes labialis, genital herpes, etc. VZV causes varicella during the primary infection, and shingles when reactivated. In this review, we present to the typical cases of each disease; in addition, we present several types of diseases that are closely related to neurological diseases.


ERK1/2 phosphorylates HIF-2α and regulates its activity by controlling its CRM1-dependent nuclear shuttling.
Hypoxia-inducible factor 2 (HIF-2) is a principal component of the cellular response to oxygen deprivation (hypoxia). Its inducible subunit, HIF-2α (also known as EPAS1), is controlled by oxygen-dependent as well as oxygen-independent mechanisms, such as phosphorylation. We show here that HIF-2α is phosphorylated under hypoxia (1% O2) by extracellular signal-regulated protein kinases 1 and 2 (ERK1/2; also known as MAPK3 and MAPK1, respectively) at serine residue 672, as identified by in vitro phosphorylation assays. Mutation of this site to an alanine residue or inhibition of the ERK1/2 pathway decreases HIF-2 transcriptional activity and causes HIF-2α to mislocalize to the cytoplasm without changing its protein expression levels. Localization, reporter gene and immunoprecipitation experiments further show that HIF-2α associates with the exportin chromosomal maintenance 1 (CRM1, also known as XPO1) in a phosphorylation-sensitive manner and identify two critical leucine residues as part of an atypical CRM1-dependent nuclear export signal (NES) neighboring serine 672. Inhibition of CRM1 or mutation of these residues restores nuclear accumulation and activity of HIF-2α lacking the ERK1/2-mediated modification. In summary, we reveal a novel regulatory mechanism of HIF-2, involving ERK1/2-dependent phosphorylation of HIF-2α, which controls its nucleocytoplasmic shuttling and the HIF-2 transcriptional activity.This article has an associated First Person interview with the first author of the paper.


Obesity paradox and cardiorespiratory fitness in 12,417 male veterans aged 40 to 70 years.
To evaluate the influence of cardiorespiratory fitness (fitness) on the obesity paradox in middle-aged men with known or suspected coronary artery disease. This study consists of 12,417 men aged 40 to 70 years (44% African American) who were referred for exercise testing at the Veterans Affairs Medical Centers in Washington, DC, or Palo Alto, CA (between January 1, 1983, and June 30, 2007). Fitness was quantified as metabolic equivalents achieved during a maximal exercise test and was categorized for analysis as low, moderate, and high (defined as <5, 5-10, and >10 metabolic equivalents, respectively). Adiposity was defined by body mass index (BMI) according to standard clinical guidelines. Separate and combined associations of fitness and adiposity with all-cause mortality were assessed by Cox proportional hazards analyses. We recorded 2801 deaths during a mean+/-SD follow-up of 7.7+/-5.3 years. Multivariate hazard ratios (95% confidence interval) for all-cause mortality, with normal weight (BMI, 18.5-24.9 kg/m2) used as the reference group, were 1.9 (1.5-2.3), 0.7 (0.7-0.8), 0.7 (0.6-0.7), and 1.0 (0.8-1.1) for BMIs of less than 18.5, 25.0 to 29.9, 30.0 to 34.9, and 35.0 or more kg/m2, respectively. Compared with highly fit normal-weight men, underweight men with low fitness had the highest (4.5 [3.1-6.6]) and highly fit overweight men the lowest (0.4 [0.3-0.6]) mortality risk of any subgroup. Overweight and obese men with moderate fitness had mortality rates similar to those of the highly fit normal-weight reference group. Fitness altered the obesity paradox. Overweight and obese men had increased longevity only if they registered high fitness.


Liver metabolism traits in two rabbit lines divergently selected for intramuscular fat.
Intramuscular fat (IMF) has a large effect in the sensory properties of meat because it affects tenderness, juiciness and flavour. A divergent selection experiment for IMF in longissimus dorsi (LD) muscle was performed in rabbits. Since liver is the major site of lipogenesis in rabbits, the objective of this work is to study the liver metabolism in the lines of the divergent selection experiment. Intramuscular fat content, perirenal fat weight, liver weight, liver lipogenic activities and plasma metabolites related to liver metabolism were measured in the eighth generation of selection. Direct response on IMF was 0.34 g/100 g of LD, which represented 2.7 SD of the trait, and selection showed a positive correlated response in the perirenal fat weight. High-IMF line showed greater liver size and greater liver lipogenic activities of enzymes glucose-6-phosphate dehydrogenase and malic enzyme. We did not find differences between lines for fatty acid synthase lipogenic activity. With regard to plasma metabolites, low-IMF line showed greater plasma concentration of triglycerides, cholesterol, bilirubin and alkaline phosphatase than high-IMF line, whereas high-IMF line showed greater albumin and alanine transaminase concentrations than low-IMF line. We did not observe differences between lines for glucose, total protein and plasma concentrations. Phenotypic correlations between fat (IMF and perirenal fat weight) and liver traits showed that liver lipogenesis affects fat deposition in both, muscle and carcass. However, the mechanisms whereby liver lipogenesis affected IMF content remain to be clarified.


Role of probiotics in urogenital healthcare.
Urogenital infections are one of the most common causes for a woman to visit a gynecologist or a urologist. The well-known association between abnormal vaginal microbial flora and its formidable risk in the increased incidence of urinary tract infection underscores the importance of understanding the microbial flora and the efforts needed to maintain it, for ensuring urogenital health. Surprisingly in spite of the increased incidence urogenital infections receive very less attention from the medical fraternity. Growing awareness among people and newer advances in the medical field has brought them into the limelight. The importance of replenishing these depleting commensals with 'probiotics' has resurfaced in a big way. As the days go by science and medicines will touch new milestones, which will include probiotics. The value of a probiotics cannot be taken at face value. Probiotics must not be considered a panacea for treating urogenital infections. However, the available data promises that it will be a strong option in improving and maintaining urogenital health.


Patterns of ecstasy use and associated harm: results of a Brazilian online survey.
The aim of this study was to gather information about ecstasy users in Brazil, particularly on issues related to risks associated to the use of the drug, so as to offer a basis to prevention projects. A total of 1,140 Brazilian ecstasy users answered an online questionnaire from August 2004 to February 2005. Participants were predominantly young single heterosexual well-educated males from upper economical classes. A categorical regression with optimal scaling (CATREG) was performed to identify the risks associated with ecstasy use. "Pills taken in life" had a significant correlation with every investigated risk, particularly ecstasy dependence, unsafe sex, and polydrug use. "Gender," "sexual orientation," and "socioeconomic class" were not predictive of risk behavior. The Internet proved to be a useful tool for data collection. Given the recent increase in ecstasy availability in Brazil, a first prevention campaign directed toward the drug is urgent. At least in a preliminary Brazilian intervention, the campaign must be conducted at night leisure places, mainly frequented by youngsters from upper socioeconomic classes. The results do not call for information material with specific targets, such as gender or sexual orientation. The study's limitations have been noted.


Radiographic and histologic evaluation of a sinus augmentation with composite bone graft: a clinical case report.
The purpose of the present study was to present a clinical case report of a sinus augmentation with a composite bone graft and delayed implant placement with a 5-year follow-up. A 40-year-old female in good health required replacement of her partial upper removable denture with a prosthesis supported by implants. Due to insufficient bone, she had to undergo a sinus lift. A block graft of autogenous chin bone, along with anorganic bovine bone covered by collagen, was utilized. Eight months postsurgery, a biopsy core was taken and 3 endosseous implants were placed. The sinus lift and implantation site are illustrated with clinical, radiological, and histological data. The discussion covers the problems raised by the choice of the surgical technique, grafting material, and immediate or delayed placement of the appropriate implant design. Our results show that a maxillary sinus augmentation with a composite bone graft is a possible method for creating adequate bone volume before implantation.


Optical Redox Imaging of Fixed Unstained Muscle Slides Reveals Useful Biological Information.
Optical redox imaging (ORI) technique images cellular autofluorescence of nicotinamide adenine dinucleotide (NADH) and oxidized flavoproteins (Fp containing FAD, i.e., flavin adenine dinucleotide). ORI has found wide applications in the study of cellular energetics and metabolism and may potentially assist in disease diagnosis and prognosis. Fixed tissues have been reported to exhibit autofluorescence with similar spectral characteristics to those of NADH and Fp. However, few studies report on quantitative ORI of formalin-fixed paraffin-embedded (FFPE) unstained tissue slides for disease biomarkers. We investigate whether ORI of FFPE unstained skeletal muscle slides may provide relevant quantitative biological information. Living mouse muscle fibers and frozen and FFPE mouse muscle slides were subjected to ORI. Living mouse muscle fibers were imaged ex vivo before and after paraformaldehyde fixation. FFPE muscle slides of three mouse groups (young, mid-age, and muscle-specific overexpression of nicotinamide phosphoribosyltransferase (Nampt) transgenic mid-age) were imaged and compared to detect age-related redox differences. We observed that living muscle fiber and frozen and FFPE slides all had strong autofluorescence signals in the NADH and Fp channels. Paraformaldehyde fixation resulted in a significant increase in the redox ratio Fp/(NADH + Fp) of muscle fibers. Quantitative image analysis on FFPE unstained slides showed that mid-age gastrocnemius muscles had stronger NADH and Fp signals than young muscles. Gastrocnemius muscles from mid-age Nampt mice had lower NADH compared to age-matched controls, but had higher Fp than young controls. Soleus muscles had the same trend of change and appeared to be more oxidative than gastrocnemius muscles. Differential NADH and Fp signals were found between gastrocnemius and soleus muscles within both mid-aged control and Nampt groups. Aging effect on redox status quantified by ORI of FFPE unstained muscle slides was reported for the first time. Quantitative information from ORI of FFPE unstained slides may be useful for biomedical applications.


Simulated evaluation of two triage scales in an emergency department in Israel.
At the time of this study, the Sheba Medical Center Emergency Department (ED) in Israel had no formal triage system in place. To evaluate the interobserver reliability of two triage scales among nurses in our ED, the time-based Australasian Triage Scale (ATS) and the resource-based Emergency Severity Index (ESI), 10 nurses participated in a workshop on ATS and ESI. They then independently assessed 100 simulated triage scenarios taken from actual ED patients, and completed a survey. Intraclass correlation coefficients were calculated. The intraclass correlation coefficient for ATS was 0.64 (95% confidence interval: 0.57, 0.71), whereas for ESI, it was 0.52 (95% confidence interval: 0.45, 0.61). The nurses felt that ESI was slightly easier to use. Using conventional interpretations, the agreement for ATS is considered substantial, whereas that for ESI is considered moderate. Conversely, the nurses found the ESI somewhat easier to use.


Microglial and astrocytic changes in the striatum of methamphetamine abusers.
Little is known about the role of glial cells in the striatum of chronic methamphetamine (METH) users. In this study, we immunohistochemically examined glial reactions in the striatum of chronic METH users who did not abstain from METH use and died of drug intoxication. Human glucose transporter 5 (hGLUT), a useful marker of microglia, and CR3.43, a major histocompatibility complex class II antigen specific for reactive microglia, were immunostained. Glial fibrillary acidic protein (GFAP) and S100 Beta were used for astrocyte immunohistochemistry. We analyzed 12 chronic METH users and 13 control subjects, and detected a 200-240% increase in the number of hGLUT5-positive cells in chronic METH users (p<0.01). However, we did not detect any proliferation of CR3.43-positive cells. The number of GFAP-positive astrocytes increased, but this increase was not significant (p>0.05). Moreover, S100B-positive cell density between the two groups was not significant (p>0.05). This study demonstrates the absence of reactive gliosis in the striatum of chronic METH users who did not abstain for prolonged periods from METH use. The results suggest that chronic METH use by itself did not activate glial cells in humans and reactive gliosis may not be involved in the mechanism underlying the loss of control in drug intake, which is a characteristic feature of drug addiction.


Maternal confidence in China: association with infant neurobehaviors but not sociodemographic variables.
To examine the relations of sociodemographic factors and infant neurobehaviors to maternal confidence in China. The Brazelton Neonatal Behavioral Assessment Scale, Family APGAR, and Maternal Confidence in Caring for the Newborn scales were administered to 40 healthy, full-term neonates. Range and regulation of state, autonomic stability, and reflex cluster scores were positively correlated; the autonomic stability cluster score was negatively correlated with maternal confidence in meeting the infant's social and instrumental needs. Educational level, age, income, satisfaction with family conditions, and infant sex were not associated with maternal confidence. Range of state and autonomic stability cluster scores predicted maternal confidence. The infant's abilities to tolerate stimuli, and to be consoled, were associated with maternal confidence. Also, maternal confidence was related to the recognition of infant autonomic cues. Family and sociodemographic variables were not associated with maternal confidence. The sex of the newborn did not affect maternal confidence.


Human chlorine gas exposition and its management - an umbrella review on human data.
Even though exposure to chlorine gas has been quite frequent in the past few decades, no specific antidotes exist. This umbrella review aimed to investigate possible recommendations for treatment after a chlorine gas exposure. A published systematic review protocol that adapted the existing Navigation Guide methodology was used for including studies without comparator. Using PubMed, Web of Science, Google scholar for all potentially relevant systematic reviews, two authors independently included papers and extracted data. The risk of bias and quality of evidence was assessed by two independent review teams blinded to each other. A qualitative summary of the study findings was conducted for this overview. There were a total of 31 studies, from 4 systematic reviews, that met the inclusion criteria, comprising 3567 reported cases, with only two studies with comparators. Six studies reported pre-hospital management of patients after exposure to chlorine gas. With respect to the treatment, the most used were oxygen therapy, endotracheal intubation, β2-agonists, and corticosteroids. This review found a high quality of evidence for the effectiveness of pre-hospital management (i.e. exposure cessation) on survival at hospital discharge after exposure to chlorine gas. Oxygen administration was effective with moderate quality of evidence, as well as other types of treatment (e.g. β2, corticosteroids), but with a low level of evidence. This umbrella review highlighted the low level of evidence for existing treatments of chlorine gas poisoning. This project was supported by the French Pays de la Loire region and Angers Loire Métropole (TEC-TOP project). There is no award/grant number. The review protocol was registered on PROSPERO under the registration number CRD42021231524.


Association of frailty and physical function in patients with non-dialysis CKD: a systematic review.
Frailty is a condition characterized by a decline in physical function and functional capacity. Common symptoms of frailty, such as weakness and exhaustion, are prevalent in patients with chronic kidney disease (CKD). The increased vulnerability of frail patients with coexisting CKD may place them at a heightened risk of encountering additional health complications. The purpose of this systematic review was to explore the link between frailty, CKD and clinical outcomes. We searched for cross sectional and prospective studies in the general population and in the CKD population indexed in EMBASE, Pubmed, Web of Science, CINAHL, Cochrane and Ageline examining the association between frailty and CKD and those relating frailty in patients with CKD to clinical outcomes. We screened 5,066 abstracts and retrieved 108 studies for full text review. We identified 7 studies associating frailty or physical function to CKD. From the 7 studies, we identified only two studies that related frailty in patients with CKD to a clinical outcome. CKD was consistently associated with increasing frailty or reduced physical function [odds ratios (OR) 1.30 to 3.12]. In patients with CKD, frailty was associated with a greater than two-fold higher risk of dialysis and/or death [OR from 2.0 to 5.88]. CKD is associated with a higher risk of frailty or diminished physical function. Furthermore, the presence of frailty in patients with CKD may lead to a higher risk of mortality. Further research must be conducted to understand the mechanisms of frailty in CKD and to confirm its association with clinical outcomes.


Comparative policies of two national dental associations: Norway and the United States.
The major issues and challenges confronting the dental professions in the United States and Norway were studied through speeches of and interviews with the presidents of the American Dental Association (ADA) and the Norwegian Dental Association (NDA) in the period 1980-86. The issues most frequently dealt with related to public authorities, particularly legislation and government involvement in dental practice. Anxiety concerning "busyness," the future dental market, and a drop in the quality of applicants to dental schools were also major subjects. The spokespersons of both associations were engaged in increasing the demand for dental services by marketing, but they were ambivalent regarding advertising, especially individual advertising. Both were concerned with protecting dentistry's autonomy. While the Norwegian presidents apparently feared the medical profession's influence upon dentistry, the Americans were concerned with the hygienists and denturists, and with the insurance companies which they suspected of intruding into the dentist-patient relationship. The presidents' statements, which frequently varied, were apparently influenced by the current domestic political climate, the basic socio-political principles of the two countries, and the different socio-demographic make-up of their memberships.


Behavior, brain, and genome in genomic disorders: finding the correspondences.
Within the last decade or so, there has been an acceleration of research attempting to connect specific genetic lesions to the patterns of brain structure and activation. This article comments on observations that have been made based on these recent data and discusses their importance for the field of investigations into developmental disorders. In making these observations, the authors focus on one specific genomic lesion, the well-studied, yet still incompletely understood, 22q11.2 deletion syndrome. The authors demonstrate the degree of variability in the phenotype that occurs at both the brain and behavioral levels of genomic disorders and describe how this variability is, on close inspection, represented at the genomic level. The authors emphasize the importance of combining genetic/genomic analyses and neuroimaging for research and for future clinical diagnostic purposes and for the purposes of developing individualized, patient-tailored treatment and remediation approaches.


Cementless total knee arthroplasty in juvenile onset rheumatoid arthritis.
Twenty-two total knee arthroplasties with at least one cementless component were performed in 14 patients with juvenile rheumatoid arthritis (JRA) from 1985 to 1989. All 22 femoral components and ten tibial components were implanted cementless. The mean age at operation was 26 years. All 14 patients were available for follow-up evaluation at an average of 3.9 years (range, two to 6.2 years). Using The Knee Society's scoring system, the knee score improved from an average of 18 points (range, 0-47 points) preoperatively to 92 points (range, 58-100 points) at follow-up evaluation. The functional score improved from 28 points (range, 0-55 points) to 76 points (range, 40-100 points). Nonprogressive radiolucencies of less than 1 mm were observed in two knees. One reoperation was performed for failure of a metal-backed patellar component. Knee arthroplasty with cementless components in selected JRA patients can give results comparable with a fully cemented knee at the two- to six-year follow-up evaluation.


[Influence of formate on free amino acids contents in tissues and organs of rats in vitro and in vivo].
In experiments in vitro and in vivo influence of formiate on free amino acids contents of tissues and organs of rats has been studied. Results of experiments showed a possibility of amino acids formilation in all investigated tissues. This process could be enzymatic. Its effectivness depended on amino acids and formiate concentrations. The most effective processes took place in organs with high metabolic activity.


Acetylglucosaminidase, an early enzyme in the development of Dictyostelium discoideum.
The specific activity of acetylglucosaminidase has been found to increase more than 10-fold during the first 10 hr of development in the cellular slime mold Dictyostelium discoideum. The specific activity then remained essentially constant until after germination. The activity was purified 36-fold and found to behave as a single protein species. The increase in specific activity required concomitant protein synthesis. If ribonucleic acid synthesis was preferentially inhibited during the period of synthesis of acetylglucosaminidase, further increase in enzymatic activity stopped after 2 hr. The increase in activity did not occur in a mutant strain which did not undergo the first step in morphogenesis. Mutant strains, blocked slightly later in morphogenesis, synthesized the enzyme at the normal rate but for an extended period. It was concluded that the initiation and termination of synthesis of acetylglucosaminidase are controlled by the developmental program.


High affinity restricts the localization and tumor penetration of single-chain fv antibody molecules.
Antitumor monoclonal antibodies must bind to tumor antigens with high affinity to achieve durable tumor retention. This has spurred efforts to generate high affinity antibodies for use in cancer therapy. However, it has been hypothesized that very high affinity interactions between antibodies and tumor antigens may impair efficient tumor penetration of the monoclonal antibodies and thus diminish effective in vivo targeting (K. Fujimori et al., J. Nucl. Med., 31: 1191-1198, 1990). Here we show that intrinsic affinity properties regulate the quantitative delivery of antitumor single-chain Fv (scFv) molecules to solid tumors and the penetration of scFv from the vasculature into tumor masses. In biodistribution studies examining a series of radioiodinated scFv mutants with affinities ranging from 10(-7)-10(-11) M, quantitative tumor retention did not significantly increase with enhancements in affinity beyond 10(-9) M. Similar distribution patterns were observed when the scFv were evaluated in the absence of renal clearance in anephric mice, indicating that the rapid renal clearance of the scFv was not responsible for these observations. IHC and IF evaluations of tumor sections after the i.v. administration of scFv affinity mutants revealed that the lowest affinity molecule exhibited diffuse tumor staining whereas the highest affinity scFv was primarily retained in the perivascular regions of the tumor. These results indicate that antibody-based molecules with extremely high affinity have impaired tumor penetration properties that must be considered in the design of antibody-based cancer therapies.


Does pain relate with activation of quadriceps and hamstrings muscles during strengthening exercise in people with knee osteoarthritis?
Muscle strengthening may be difficult to achieve in knee osteoarthritis (OA) due to pain. A large knee adduction moment (KAM), representing medial relative to lateral knee load, may also relate with pain during strengthening exercise. The objective of this study was to examine relationships between knee pain status and electromyography (EMG) amplitude of knee muscles during squat and lunge exercises. We also evaluated relationships between pain and KAM during these exercises. Forty-two women with symptomatic knee OA participated. Knee pain intensity and frequency were captured with two reliable and valid questionnaires. Motion analyses of squat and lunge exercises were completed. Total average EMG amplitude across five muscles of the lower limb and average KAM were calculated from the static portion of these exercises. Multiple regression analyses examined the relationships between pain and total average EMG amplitude; and pain and average KAM during squats and lunges. Pain improved the model for KAM from the trailing leg of a lunge. Pain did not improve any other model. Overall, pain may not be a useful indicator of EMG amplitude or KAM during exercise in knee OA.


Fission yeast leucine-rich repeat protein Lrp1 is essential for cell morphogenesis as a component of the morphogenesis Orb6 network (MOR).
In eukaryotes, cell morphogenesis is regulated coordinately with the cell cycle. In fission yeast, the morphogenesis network MOR (morphogenesis Orb6 network) consists of 5 conserved proteins, Pmo25, Nak1, Mor2, Orb6, and Mob2, and is essential for cell polarity control and cell separation following cytokinesis. Here we show that the conserved leucine-rich repeat protein Lrp1 is required for cell morphogenesis as a newly recognized component of MOR. Lrp1 has 4 leucine-rich repeats in its N-terminus and is a homolog of the budding yeast Sog2, which is a component of the RAM network (regulation of Ace2 activity and cellular morphogenesis). Lrp1 was essential for both cell growth and cell morphogenesis as were the other MOR components. Lrp1 was localized to the SPBs (spindle pole bodies, the yeast equivalent of the animal centrosome) throughout the cell cycle and to the medial ring during cytokinesis. Lrp1 interacted with Nak1 and was important for Orb6 kinase activity. Thus Lrp1 proved to function upstream of Orb6 in cell morphogenesis.


A Mobile Stroke Treatment Unit for Field Triage of Patients for Intraarterial Revascularization Therapy.
Favorable outcomes in intraarterial therapy (IAT) for acute ischemic stroke (AIS) are related to early vessel recanalization. The mobile stroke treatment unit (MSTU) is an on-site, prehospital, treatment team, laboratory, and CT scanner that reduces time to treatment for intravenous thrombolysis and may also shorten time to IAT. Using our MSTU database, we identified patients that underwent IAT for AIS. We compared the key time metrics to historical controls, which included patients that underwent IAT at our institution six months prior to implementation of the MSTU. We further divided the controls into two groups: (1) transferred to our institution for IAT and (2) directly presented to our emergency room and underwent IAT. After 164 days of service, the MSTU transported 155 patients of which 5 underwent IAT. We identified 5 historical controls that were transferred to our center for IAT. Substantial reduction in times including median door to initial CT (12 minute vs. 32 minute), CT to IAT (82 minute vs. 165 minute), and door to MSTU/primary stroke center departure (37 minute vs. 106 minute) were noted among the two groups. Compared to the 6 patients who presented to our institution directly, the MSTU process times were also shorter. Our initial experience shows that MSTU may help in early triage and shorten the time to IAT for AIS.


Effectiveness of an occupational therapy home programme in Spain for people affected by stroke.
The main aim of this paper is to assess the effects of an occupational therapy home programme in Spain on 23 people who had had stroke (mean age 61.17 years). The programme was made up of a set of activities and techniques of physical, cognitive, social and functional nature aimed at preventing, maintaining and/or rehabilitating the abilities affected of people who had had stroke. A multiple-baseline intrasubject design and replication with a treatment withdrawal period to check whether the effects of the programme remained was applied. The results show a significant statistical improvement, concerning not only the participants' cognitive skills through Loewenstein Occupational Therapy Cognitive Assessment Battery - Second Edition but also their functional independence as assessed by the Barthel Index. Despite the low number of participants, being one of the limitations of our study, the results support the need to carry out research about the effectiveness of rehabilitation treatments in the home with the goal to plan how public healthcare systems should tackle them and how to improve those already being used.


Recognition of nonexplicitly presented odors.
This research was designed to check to what extent naive subjects can recognize odors in a situation similar to those of daily life. The odors were presented nonexplicitly and without any warning to memorize them for a later recognition task. Different intensities of noise were used to verify whether these affect the olfactory recognition task by modifying arousal and check whether a situation made more unpleasant by a louder noise produces changes in subjects' evaluations of the pleasantness of the odors presented. Also, subjects' personality traits were measured on extraversion-introversion and self-monitoring questionnaires. The three odors (Eucalyptolus, linalile acetate, bornile acetate) were recognized just under 50% of the time and were recognized differently. The number of recognitions did not vary significantly by sex or noise conditions; the most intense noise affected only the number of false alarms. Pleasantness scores differed only among the different odors. High self-monitoring subjects tended to have fewer recognitions; scores on extraversion-introversion produced no significant effect.