The mechanism of intestinal uptake and transcellular transport of IgG in the neonatal rat.
The transport of immunoglobulins across the intestinal mucosa of neonatal rats provides an excellent model for the study of transcellular protein transport. The mechanism of intestinal uptake and transcellular transport of plasma proteins has been studied in 12-14-day old rats using intraduodenally administered radioiodinated proteins. Appreciable quantities of rat IgG, mouse IgG, rabbit IgG, and all four subclasses of human IgG were taken up by the intestinal wall (19-54% of administered dose at 4 hr) and transported to the animal (10-35% of administered dose at 4 hr). In contrast there was little or no uptake of human IgM, IgA, and IgE and little or no transport of human IgM, IgA, IgD, IgE, albumin, transferrin, and ceruloplasmin. Both the uptake and transport of labeled IgG were significantly inhibited by unlabeled IgG. Further insight into the transport process was obtained from the observation that an appreciable proportion of the label of IgG in intestinal wall homogenates, but not in plasma or intestinal washings, migrated in a sucrose ultracentrifugation gradient much more rapidly than did the administered 7S molecules. This pattern was not observed with other proteins studied. This apparent binding of labeled IgG was also markedly inhibited by unlabeled IgG. In subcellular fractionation studies of intestinal homogenates the complexed labeled IgG was shown to be associated predominantly with cell membrane rather than cell sap fractions. In addition IgG could be shown to bind to purified enterocyte microvillous membranes in vitro. IT IS CONCLUDED THAT IN THE NEONATAL RAT: (a) the major processes involved in both intestinal uptake and transport of IgG are specific and saturable; (b) intestinal transport is associated with complexing of IgG molecules with membranes, most probably with enterocyte microvillous membranes; and (c) the part of the IgG structure involved in this process is probably similar to that involved in the concentration-catabolism effect but is not identical to that mediating other non-antigen combining functions of IgG. Our data are consistent with the existence of specific receptors for IgG on enterocyte microvillous membranes of the neonatal rat. Such receptors would be necessary for the specific uptake and transport of these molecules.


Regulation of miRNA abundance by RNA binding protein TOUGH in Arabidopsis.
MicroRNAs (miRNAs) are regulators of gene expression in plants and animals. The biogenesis of miRNAs is precisely controlled to secure normal development of organisms. Here we report that TOUGH (TGH) is a component of the DCL1-HYL1-SERRATE complex that processes primary transcripts of miRNAs [i.e., primary miRNAs (pri-miRNAs)] into miRNAs in Arabidopsis. Lack of TGH impairs multiple DCL activities in vitro and reduces the accumulation of miRNAs and siRNAs in vivo. TGH is an RNA-binding protein, binds pri-miRNAs and precursor miRNAs in vivo, and contributes to pri-miRNA-HYL1 interaction. These results indicate that TGH might regulate abundance of miRNAs through promoting DCL1 cleavage efficiency and/or recruitment of pri-miRNAs.


Induced Pluripotent Stem Cell-Derived Megakaryocytes and Platelets for Disease Modeling and Future Clinical Applications.
Platelets, derived from megakaryocytes, are anucleate cytoplasmic discs that circulate in the blood stream and play major roles in hemostasis, inflammation, and vascular biology. Platelet transfusions are used in a variety of medical settings to prevent life-threatening thrombocytopenia because of cancer therapy, other causes of acquired or inherited thrombocytopenia, and trauma. Currently, platelets used for transfusion purposes are donor derived. However, there is a drive to generate nondonor sources of platelets to help supplement donor-derived platelets. Efforts have been made by many laboratories to generate in vitro platelets and optimize their production and quality. In vitro-derived platelets have the potential to be a safer, more uniform product, and genetic manipulation could allow for better treatment of patients who become refractory to donor-derived units. This review focuses on potential clinical applications of in vitro-derived megakaryocytes and platelets, current methods to generate and expand megakaryocytes from pluripotent stem cell sources, and the use of these cells for disease modeling.


Modulation and phosphorylation of calbindin-D28K correlates with protein kinase C activation1.
Calbindin-D28K is a vitamin D3 dependent calcium-binding protein expressed in renal distal tubules. We previously reported that 24 h treatment with 1 alpha, 25-dihydroxyvitamin D3 (1,25-D3), the active form of vitamin D3, induces calbindin-D28K and activates protein kinase C (PKC) in MDBK (Madin-Darby bovine kidney) cells. In contrast, 24 h treatment with the phorbol ester 12-O-tetradecanoylphorbol-3-acetate (TPA) downregulates calbindin-D28K and PKC activity. In the present studies, we demonstrate that TPA rapidly enhances calbindin-D28K expression in MDBK cells in the absence of 1,25-D3. The enhancement of calbindin-D28K expression is preceded by activation and translocation of PKC alpha. Further, we show that PKC directly phosphorylates calbindin-D28K in a calcium- and phospholipid-dependent manner in vitro. In MDBK cells, the calbindin-D28K antibody immunoprecipitates a 28 kDa protein for which phosphorylation is enhanced after treatment for 1 h with TPA or 24 h with 1,25-D3. Consistent with amino acid sequence analysis of calbindin-D28K indicating two threonine residues that fit the consensus for PKC phosphorylation, TPA-treated MDBK cells exhibit enhanced expression of a phosphothreonine-containing protein that co-migrates with calbindin-D28K. These studies offer the first report that calbindin-D28K is a phosphoprotein and implicate the PKC signal transduction pathway in its regulation.


Transcriptional response of dermal fibroblasts in direct current electric fields.
During the course of normal wound healing, fibroblasts at the wound edge are exposed to electric fields (EFs) ranging from 40 to 200 mV/mm. Various forms of EFs influence fibroblast migration, proliferation, and protein synthesis. Thus, EFs may contribute to fibroblast activation during wound repair. To elucidate the role of EFs during the normal progression of healing, this study compares gene expression in normal adult dermal fibroblasts exposed to a 100 mV/mm EF for 1 h to non-stimulated controls. Significantly increased expression of 162 transcripts and decreased expression of 302 transcripts was detected using microarrays, with 126 transcripts above the level of 1.4-fold increases or decreases compared to the controls. Above the level of twofold, only 11 genes were significantly increased or decreased compared to controls. Many of these significantly regulated genes are associated with wound repair through the processes of matrix production, cellular signaling, and growth. Activity within specific cellular signaling pathways is noted, including TGF-beta, G-proteins, and inhibition of apoptosis. In addition, RT-PCR analysis of the expression of KLF6, FN1, RGS2, and JMJD1C over continued stimulation and at different field strengths suggests that there are specific windows of field characteristics for maximum induction of these genes. EFs thus appear to have an important role in controlling fibroblast activity in the process of wound healing.


Chemical construction and structural permutation of potent cytotoxin polytheonamide B: discovery of artificial peptides with distinct functions.
Polytheonamide B (1), isolated from the marine sponge Theonella swinhoei, is a posttranslationally modified ribosomal peptide (MW 5030 Da) that displays extraordinary cytotoxicity. Among its 48 amino acid residues, this peptide includes a variety D- and L-amino acids that do not occur in proteins, and the chiralities of these amino acids alternate in sequence. These structural features induce the formation of a stable β6.3-helix, giving rise to a tubular structure of over 4 nm in length. In the biological setting, this fold is believed to transport cations across the lipid bilayer through a pore, thereby acting as an ion channel. In this Account, we discuss the construction and structural permutations of this potent cytotoxin. First we describe the 161-step chemical construction of this unusual peptide 1. By developing a synthetic route to 1, we established the chemical basis for subsequent SAR studies to pinpoint the proteinogenic and nonproteinogenic building blocks within the molecule that confer its toxicity and channel function. Using fully synthetic 1, we generated seven analogues with point mutations, and studies of their activity revealed the importance of the N-terminal moiety. Next, we simplified the structure of 1 by substituting six amino acid residues of 1 to design a more synthetically accessible analogue 9. This dansylated polytheonamide mimic 9 was synthesized in 127 total steps, and we evaluated its function to show that it can emulate the toxic and ion channel activities of 1 despite its multiple structural modifications. Finally, we applied a highly automated synthetic route to 48-mer 9 to generate 13 substructures of 27-39-mers. The 37-mer 12 exhibited nanomolar level toxicity through a potentially distinct mode of action from 1 and 9. The SAR studies of polytheonamide B and the 21 artificial analogues have deepened our understanding of the precise structural requirements for the biological functions of 1. They have also led to the discovery of artificial molecules with various toxicities and channel functions. These achievements demonstrate the benefits of total synthesis and the importance of efficient construction of complex molecules. The knowledge accumulated through these studies will be useful for the rational design of new tailor-made channel peptides and cytotoxic molecules with desired functions.


Recombination drives genetic diversification of Streptococcus dysgalactiae subspecies equisimilis in a region of streptococcal endemicity.
Infection of the skin or throat by Streptococcus dysgalactiae subspecies equisimilis (SDSE) may result in a number of human diseases. To understand mechanisms that give rise to new genetic variants in this species, we used multi-locus sequence typing (MLST) to characterise relationships in the SDSE population from India, a country where streptococcal disease is endemic. The study revealed Indian SDSE isolates have sequence types (STs) predominantly different to those reported from other regions of the world. Emm-ST combinations in India are also largely unique. Split decomposition analysis, the presence of emm-types in unrelated clonal complexes, and analysis of phylogenetic trees based on concatenated sequences all reveal an extensive history of recombination within the population. The ratio of recombination to mutation (r/m) events (11:1) and per site r/m ratio (41:1) in this population is twice as high as reported for SDSE from non-endemic regions. Recombination involving the emm-gene is also more frequent than recombination involving housekeeping genes, consistent with diversification of M proteins offering selective advantages to the pathogen. Our data demonstrate that genetic recombination in endemic regions is more frequent than non-endemic regions, and gives rise to novel local SDSE variants, some of which may have increased fitness or pathogenic potential.


Characterizing Drug-Polymer Interactions in Aqueous Solution with Analytical Ultracentrifugation.
We present a new approach for characterizing drug-polymer interactions in aqueous media, using sedimentation velocity analytical ultracentrifugation (AUC). We investigated the potential interaction of ketoconazole (KTZ), a poorly water-soluble drug, with polyacrylic acid (PAA) and a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus) in aqueous buffers. The effect of the polymer on the sedimentation coefficient of the drug was the observable metric. The drug alone, when subjected to AUC, exhibited a very narrow sedimentation peak at 0.2 Svedberg (S), in agreement with the expectation for a monomeric drug with a molar mass < 1000 Dalton. Conversely, the neat polymers showed broad profiles with higher sedimentation coefficients, reflecting their larger more heterogeneous size distributions. The sedimentation profiles of the drug-polymer mixtures were expectedly different from the profile of the neat drug. With KTZ-Soluplus, a complete shift to faster sedimentation times (indicative of an interaction) was observed, while with KTZ-PAA, a split peak indicated the existence of the drug in both free and interacting states. The sedimentation profile of carbamazepine, a second model drug, in the presence of hydroxypropyl methyl cellulose acetate succinate (HPMCAS, another polymer) revealed multiple "populations" of drug-polymer species, very similar to the sedimentation profile of neat HPMCAS. The interactions probed by AUC were compared with the results from isothermal titration calorimetry. In vitro dissolution tests performed on amorphous solid dispersions prepared with the same drug-polymer pairs suggested that the interactions may play a role in prolonging drug supersaturation. The results show the possibility of characterizing drug-polymer interactions in aqueous solution with high hydrodynamic resolution, addressing a major challenge frequently encountered in the mechanistic investigations of the dissolution behavior of amorphous solid dispersions.


Living with achondroplasia: quality of life evaluation following cervico-medullary decompression.
Achondroplasia is the most common of the heritable skeletal dysplasias. Cervico-medullary compression is a frequently encountered and potentially lethal neurological complication. Cervico-medullary decompression (CMD) at the foramen magnum is often employed to relieve the pressure on the emerging cervical cord. Given the inherent risks associated with major surgery, there has been a substantial debate regarding the best criteria for CMD. Our objectives for this study are to explore the quality of life of patients who had undergone CMD, and to assess whether surgery is associated with mortality and increased long-term morbidity. A Medical Outcome Study 36-item Short Form General Health Survey designed to evaluate eight general health concepts as well as achondroplasia-related issues, was administered to patients assessed in the neurosurgery department in Johns Hopkins Hospital between 1977 and 1998. One hundred and sixty-seven patients were eligible for inclusion. Forty-three could not be contacted, and two refused consent. One hundred and twenty-two patients were assessed. Fifty-six (46%) individuals had CMD and 66 (54%) did not. There was 1 case of mortality in the CMD group and 12 cases in the non-CMD group. In the non-CMD group, all deaths, as far as we know, were unrelated to cervico-medullary compression. In this cohort of surviving patients (n = 109), the quality of life of the 55 (50.5%) who had undergone CMD is comparable to that of the 54 (49.5%) who did not have surgery, controlled for age and sex. CMD is indicated for patients with achondroplasia with significant symptomatic foramen magnum compression. It can be life saving. It can abolish profound central apnea that may cause sudden death and alleviate neurological complications associated with damage of the significantly compressed spinal cord. With regards to long-term outcome evaluation, the quality of life of individuals with achondroplasia who had CMD is similar to those age- and sex-matched patients who did not have this surgery. Moreover, CMD, with all its inherent surgical risks, does not appear to be associated with higher mortality or increased long-term morbidity.


Inter-rater variability as mutual disagreement: identifying raters' divergent points of view.
Whenever multiple observers provide ratings, even of the same performance, inter-rater variation is prevalent. The resulting 'idiosyncratic rater variance' is considered to be unusable error of measurement in psychometric models and is a threat to the defensibility of our assessments. Prior studies of inter-rater variation in clinical assessments have used open response formats to gather raters' comments and justifications. This design choice allows participants to use idiosyncratic response styles that could result in a distorted representation of the underlying rater cognition and skew subsequent analyses. In this study we explored rater variability using the structured response format of Q methodology. Physician raters viewed video-recorded clinical performances and provided Mini Clinical Evaluation Exercise (Mini-CEX) assessment ratings through a web-based system. They then shared their assessment impressions by sorting statements that described the most salient aspects of the clinical performance onto a forced quasi-normal distribution ranging from "most consistent with my impression" to "most contrary to my impression". Analysis of the resulting Q-sorts revealed distinct points of view for each performance shared by multiple physicians. The points of view corresponded with the ratings physicians assigned to the performance. Each point of view emphasized different aspects of the performance with either rapport-building and/or medical expertise skills being most salient. It was rare for the points of view to diverge based on disagreements regarding the interpretation of a specific aspect of the performance. As a result, physicians' divergent points of view on a given clinical performance cannot be easily reconciled into a single coherent assessment judgment that is impacted by measurement error. If inter-rater variability does not wholly reflect error of measurement, it is problematic for our current measurement models and poses challenges for how we are to adequately analyze performance assessment ratings.


Does the Illness Behavior Questionnaire measure abnormal illness behavior?
Abnormal illness behavior (AIB) has been proposed as a construct measuring the inappropriate or maladaptive modes of responding to one's state of health, and the Illness Behavior Questionnaire (BQ; Pilowsky, 1975) was designed to measure this construct. Previous studies using small samples have failed to agree on the factor structure of this questionnaire. The present paper examines the factor structure of the Illness Behavior Questionnaire and critically evaluates the interpretation of its dimensions as well as the construct of AIB. A factor analysis of responses from 1,061 health care and nonhealth care seeking subjects yielded six interpretable factors which substantially replicated Pilowsky's previous results. Six scales were calculated and correlated with several personality measures. The results indicated that the Illness Behavior Questionnaire is saturated with neuroticism, a dimension known to be related to excessive medical complaints. But excessive medical complaints cannot be equated with hypochondriasis or AIB in the absence of objective medical information. In the absence of evidence for the discriminant validity of the IBQ, its use as a diagnostic device is unwarranted. Treating elevated IBQ scores as indicators of abnormal illness behavior without corroborating medical information may be more misleading than accepting patients' symptom reports at face value.


Detection of a Cooper-pair density wave in Bi2Sr2CaCu2O8+x.
The quantum condensate of Cooper pairs forming a superconductor was originally conceived as being translationally invariant. In theory, however, pairs can exist with finite momentum Q, thus generating a state with a spatially modulated Cooper-pair density. Such a state has been created in ultracold (6)Li gas but never observed directly in any superconductor. It is now widely hypothesized that the pseudogap phase of the copper oxide superconductors contains such a 'pair density wave' state. Here we report the use of nanometre-resolution scanned Josephson tunnelling microscopy to image Cooper pair tunnelling from a d-wave superconducting microscope tip to the condensate of the superconductor Bi2Sr2CaCu2O8+x. We demonstrate condensate visualization capabilities directly by using the Cooper-pair density variations surrounding zinc impurity atoms and at the Bi2Sr2CaCu2O8+x crystal supermodulation. Then, by using Fourier analysis of scanned Josephson tunnelling images, we discover the direct signature of a Cooper-pair density modulation at wavevectors QP ≈ (0.25, 0)2π/a0 and (0, 0.25)2π/a0 in Bi2Sr2CaCu2O8+x. The amplitude of these modulations is about five per cent of the background condensate density and their form factor exhibits primarily s or s' symmetry. This phenomenology is consistent with Ginzburg-Landau theory when a charge density wave with d-symmetry form factor and wavevector QC = QP coexists with a d-symmetry superconductor; it is also predicted by several contemporary microscopic theories for the pseudogap phase.


Racial differences in vascular risk factors and outcomes of patients with intracranial atherosclerotic arterial stenosis.
Atherosclerotic intracranial stenosis is an important cause of stroke in blacks, yet there are limited data on vascular risk factors and outcome. We analyzed the vascular risk factors and outcomes of blacks and whites in the Warfarin versus Aspirin for Symptomatic Intracranial Disease (WASID) trial. Baseline characteristics and outcomes (ischemic stroke, brain hemorrhage, or vascular death combined and ischemic stroke alone) were compared between blacks (n=174) and whites (n=331) using univariate and multivariate analyses. Blacks were significantly (P<0.05) more likely than whites to be/have: female, hypertension history, diabetes history, higher LDL, higher total cholesterol, lower triglycerides, unmarried, unemployed, nonprivate insurance, no insurance, stroke as qualifying event, <70% stenosis, symptomatic anterior circulation vessel, no antithrombotic medication before qualifying event, and no family history of myocardial infarction. Blacks more frequently reached an end point of ischemic stroke, brain hemorrhage or vascular death (28% versus 20%; hazard ratio of 1.49, 95% CI 1.03 to 2.17, P=0.03), had a higher 2-year event rate (0.28 versus 0.19), and reached the end point of ischemic stroke alone (25% versus 16% at 2 years; hazard ratio of 1.62, P=0.017). In multivariate analysis, race was associated with ischemic stroke (P=0.0488) but not with the end point ischemic stroke, brain hemorrhage or vascular death (P=0.188). Blacks with intracranial stenosis are at higher risk of stroke recurrence than whites. This risk warrants additional study of factors contributing to stroke in blacks and highlights the need for aggressive risk factor management in blacks to prevent recurrence.


Fucoidan protects mesenchymal stem cells against oxidative stress and enhances vascular regeneration in a murine hindlimb ischemia model.
Mesenchymal stem cells (MSCs) have the potential to differentiate into multiple cell lineages. Given this potential for tissue regeneration, MSC-based therapeutic applications have been considered in recent years. However, ischemia-induced apoptosis has been reported to be one of the main causes of MSC death following transplantation. The primary objective of this study was to determine whether a natural antioxidant, fucoidan, could protect MSCs from ischemia-induced apoptosis in vitro and in vivo. Furthermore, we investigated the mechanism of action of fucoidan's anti-ischemic effect in MSCs. Pre-treatment with fucoidan (10 μg/mL) suppressed the increase in H2O2-induced reactive oxygen species (ROS) levels and drastically reduced apoptotic cell death in MSCs. Fucoidan inhibited the activation of the pro-apoptotic proteins p38-mitogen-activated protein kinase (MAPK), Jun N-terminal kinase (JNK), and caspase-3, and augmented the expression of the anti-apoptosis protein cellular inhibitor of apoptosis (cIAP). Moreover, fucoidan significantly increased manganese superoxide dismutase (MnSOD) expression and decreased cellular ROS levels via the Akt pathway, resulting in enhanced cell survival. In a murine hindlimb ischemia model, transplanted fucoidan-treated MSCs showed significantly enhanced cell survival and proliferation in ischemic tissues. Functional recovery and limb salvage also remarkably improved in mice injected with fucoidan-stimulated MSCs compared with mice injected with non-stimulated MSCs. Taken together, these results show that fucoidan protects MSCs from ischemia-induced cell death by modulation of apoptosis-associated proteins and cellular ROS levels through regulation of the MnSOD and Akt pathways, suggesting that fucoidan could be powerful therapeutic adjuvant for MSC-based therapy in ischemic diseases.


Analysis of DNA repair-related genes in breast cancer reveals CUL4A ubiquitin ligase as a novel biomarker of trabectedin response.
Trabectedin is more active in nucleotide excision repair (NER)-efficient and homologous recombination repair (HRR)-deficient cells. As up to 25% of sporadic breast tumors present somatic inactivation of the HRR pathway (BRCAness phenotype), we sought to characterize trabectedin effect in BRCA1-proficient and BRCA1-null breast cancer cell lines. We evaluated whether HRR and NER gene expression correlates with trabectedin sensitivity and explored the response predictive value of the CUL4A ubiquitin ligase, which ubiquitinates NER pathway members. We characterized trabectedin cytotoxicity, cell-cycle effects, and BRCA1, BRCA2, XRCC3, XPG, ERCC1, and CUL4A expression in 10 breast cancer cell lines. Gene expression and trabectedin sensitivity association were determined in cell lines. Survival assays after trabectedin treatment were conducted in CUL4A-silenced BRCA1-proficient and -deficient cells. Because of limited phase II clinical trials evaluating trabectedin efficacy in patients with breast cancer, we assessed CUL4A immunohistochemical staining in a retrospective series of 118 sarcomas from trabectedin-treated patients to validate in vivo our in vitro observations. In cell lines, greater trabectedin sensitivity was associated with higher CUL4A expression and lower BRCA1/ERCC5, BRCA1/CUL4A, and XRCC3/CUL4A expression ratios. In agreement, BRCA1-deficient CUL4A-knockdown cells presented higher cell survival after trabectedin exposure than did scramble control cells. Lack of effect in BRCA1-proficient cells suggests that HRR impairment is key in CUL4A-mediated trabectedin sensitivity. High CUL4A expression in nontranslocation-related patients with sarcoma predicted improved progression-free survival [PFS; HR, 0.37; 95% confidence interval (CI), 0.20-0.68, P = 0.001] and overall survival (OS; HR, 0.44; 95% CI, 0.21-0.93, P = 0.026). Our observations support the notion of greater trabectedin activity in tumors exhibiting BRCAness and reveal CUL4A as a potential biomarker for definition of trabectedin target patients.


The ADVANCE study: a groundbreaking trial to evaluate a candidate universal antiretroviral regimen.
Current WHO-recommended first-line therapy in low-income and middle-income countries has been very successful in saving millions of lives but still has toxicity concerns and a low barrier to resistance. Two candidate antiretrovirals may substantially transform first-line therapy in low-income and middle-income countries, yielding a safer, more robust and cheaper regimen. Tenofovir alafenamide carries toxicity and cost benefits over tenofovir disoproxil fumarate. Dolutegravir could replace efavirenz, with substantial toxicity, resistance and cost benefits. However, these drugs are currently not manufactured together in developed countries, for commercial reasons. We describe a large randomized controlled study testing a combination of these candidate antiretrovirals against the current first-line recommendation that commenced recruitment in early 2017. We justify the study design and discuss how we will deal with complex issues such as tuberculosis and pregnancy.


Production of IL-1Ra by human mononuclear blood cells in vitro: influence of serum factors.
In vitro cell culture models that measure cytokine production can be of great value when analyzing regulatory mechanisms underlying various pathological conditions. However, testing the function of peripheral blood cells has to take into consideration that serum factors are likely to be of importance in maintaining their function. Interleukin-1 receptor antagonist (IL-1Ra) is a cytokine of key importance in immune regulation and is believed to be involved in numerous pathological processes, such as autoimmunity and cancer. We investigated the influence of normal, human serum on spontaneous production of IL-1Ra by human peripheral blood mononuclear cells (PBMC) in vitro. IL-1Ra production in vitro spanned over a wide range of concentrations, which could be attributed to a combined effect of both cellular parameters and properties of the serum used. The production of IL-1Ra in vitro could be correlated to the level of immobilized IgG, especially IgG1 and IgG3, which is adsorbed from the serum and bound to the tissue culture wells during culture. However, the amount of serum IgG adsorbed to the tissue culture wells could not necessarily be predicted based on the serum concentration of IgG.


Oxygraphic evaluation of mitochondrial function in digitonin-permeabilized mononuclear cells and cultured skin fibroblasts of patients with chronic progressive external ophthalmoplegia.
For quantitative elucidation of maximal mitochondrial oxidation capacities in human mononuclear cells, cultured human skin fibroblasts and human thrombocytes the optimal amount of digitonin for plasma membrane permeabilization was determined to be 5, 10, and 0.1 micrograms/10(6) cells, respectively. Using these concentrations the rate of respiration of permeabilized cells with the mitochondrial substrates succinate (+ rotenone) or glutamate + malate can be stimulated between two- and fourfold by ADP and inhibited by carboxyatractyloside. The maximal respiratory activities of well-characterized preparations of permeabilized mononuclear cells of five patients with chronic progressive external ophthalmoplegia were compared to healthy controls and a 30 to 50% decrease of the ADP-stimulated respiration rates with glutamate + malate and succinate + rotenone was detected. This is an indication for the presence of the mitochondrial defect in respiratory active blood cells. Additionally, for two of these patients the mitochondrial defects were proven to be detectable by the determination of maximal oxygen consumption rates of digitonin-permeabilized cultured skin fibroblasts. Therefore, the determination of maximal oxidation capacities of a well-defined cell population using strictly standardized conditions of digitonin permeabilization is judged as a useful and sensitive method for the elucidation of mitochondrial function in extramuscular tissue.


An estimation of the artisanal small-scale production of gold in the world.
The increase in gold price of over 400% between 2002 and 2012, due to a shift towards safe investments in a period of crisis in the global economy, created a rapid increase in gold production. A response to this shift in production was observed for artisanal and small-scale mining (ASM) units in remote locations of the world, but this phenomenon has not been quantified yet. The work presented here was done to provide a quantitative tool for estimation of the gold (Au) produced by ASM and the population of workers involved in the production process, and assessment of mercury (Hg) consumed. The following hypotheses were addressed: i) It is possible to estimate, on first approximation, the amount of Au production in the world by artisanal mining; ii) Au production by artisanal mining varies by country and continent and iii) Hg consumption due to ASM can be correlated with the methods applied in the different countries and continents for the production of Au. To do this we estimated the number of miners, calculated the change in Au price and production and then applied an adjustment factor to calculate Hg production by country and continent. The amount of Au produced depends on technology of the miners by continents (highest in South America, medium in Asia and Central America, and lowest in Africa), and the geologic setting (not investigated here). The results of the estimation show that, as of 2011, over 16 million Artisanal Miners, in the world, were involved in gold extraction (mining or treatment), producing between 380 and 450 t of gold per year, with clear global behavior between the continents in terms of recovery efficiency, confirmed by data on Hg release that is higher in countries with lower technology.


Physical growth of cebus monkeys (Cebus albifrons) during the first year of life.
Infannt cebus monkeys, removed from their mothers shortly after birth, were reared in a primate nursery on diets of controlled nutritional quality. At regular intervals between birth and one year of age, each monkey was anesthetized, measured, and radiographed. Measurements were fitted to functions of the animal's age in days; a linear function for the first 6 to 8 weeks and a logarithmic function for the remainder of the first year. Mean constant curves have been calculated for each measure and estimates of animal variability have been obtained by interpolating sizes at given ages from regression lines fitted to the data for each animal. The maximum rate of growth was attained soon after birth. Cranial-caudal and distal-proximal maturity gradients in size attainment were observed.


Survival of adult generated hippocampal neurons is altered in circadian arrhythmic mice.
The subgranular zone of the hippocampal formation gives rise to new neurons that populate the dentate gyrus throughout life. Cells in the hippocampus exhibit rhythmic clock gene expression and the circadian clock is known to regulate the cycle of cell division in other areas of the body. These facts suggest that the circadian clock may regulate adult neurogenesis in the hippocampus as well. In the present study, neurogenesis in the hippocampal subgranular zone was examined in arrhythmic Bmal1 knockout (-KO) mice and their rhythmic heterozygous and wildtype littermates. Proliferation and survival of newly generated subgranular zone cells were examined using bromodeoxyuridine labelling, while pyknosis (a measure of cell death) and hippocampal volume were examined in cresyl violet stained sections. There was no significant difference in cellular proliferation between any of the groups, yet survival of proliferating cells, 6 weeks after the bromodeoxyuridine injection, was significantly greater in the BMAL1-KO animals. The number of pyknotic cells was significantly decreased in Bmal1-KO animals, yet hippocampal volume remained the same across genotypes. These findings suggest that while a functional circadian clock is not necessary for normal proliferation of neuronal precursor cells, the normal pruning of newly generated neurons in the hippocampus may require a functional circadian clock.


Radial diffusion and penetration of gas molecules and aerosol particles through laminar flow reactors, denuders, and sampling tubes.
Flow reactors, denuders, and sampling tubes are essential tools for many applications in analytical and physical chemistry and engineering. We derive a new method for determining radial diffusion effects and the penetration or transmission of gas molecules and aerosol particles through cylindrical tubes under laminar flow conditions using explicit analytical equations. In contrast to the traditional Brown method [Brown, R. L. J. Res. Natl. Bur. Stand. (U. S.) 1978, 83, 1-8] and CKD method (Cooney, D. O.; Kim, S. S.; Davis, E. J. Chem. Eng. Sci. 1974, 29, 1731-1738), the new approximation developed in this study (known as the KPS method) does not require interpolation or numerical techniques. The KPS method agrees well with the CKD method under all experimental conditions and also with the Brown method at low Sherwood numbers. At high Sherwood numbers corresponding to high uptake on the wall, flow entry effects become relevant and are considered in the KPS and CKD methods but not in the Brown method. The practical applicability of the KPS method is demonstrated by analysis of measurement data from experimental studies of rapid OH, intermediate NO3, and slow O3 uptake on various organic substrates. The KPS method also allows determination of the penetration of aerosol particles through a tube, using a single equation to cover both the limiting cases of high and low deposition described by Gormley and Kennedy (Proc. R. Ir. Acad., Sect. A. 1949, 52A, 163-169). We demonstrate that the treatment of gas and particle diffusion converges in the KPS method, thus facilitating prediction of diffusional loss and penetration of gases and particles, analysis of chemical kinetics data, and design of fluid reactors, denuders, and sampling lines.


Associations between visual hallucinations and impaired visuo-spatial abilities in dementia with Lewy bodies.
Objectives: In dementia with Lewy bodies (DLB) recurrent visual hallucinations (VH) often coexist or occur consecutively to impaired visual perception. Since in-depth neuropsychological testing is time-consuming, and therefore, not routinely performed in clinical settings, we aimed to explore whether standard cognitive screening tests may be helpful to alert a clinician to the presence of VH in DLB by exploring association between visuo-spatial dysfunction and VH. Method: The clock drawing, cube, and pentagons copying items from Montreal Cognitive Assessment and Mini-Mental State Exam and nonmotor Hooper visual organization test (HVOT) have been scored in DLB patients with and without VH using traditional and extended scoring methods. Results: Forty-five of 69 (65%) DLB patients were VH-positive (VH+). VH+ patients performed worse on the clock drawing (8.8/16 vs. 11.9/16, p = .016) with a higher rate of misrepresentation of time (69% vs. 29%, p = .002) and numbers (53% vs. 25%, p = .024). Likewise, VH+ patients performed worse on the HVOT (13.3/30 vs. 15.7/30, p = .009) having more isolated (6.2/30 vs. 4.4/30, p = .012) types of responses compared to VH- patients. Both groups had similar copying ability (p > .05). The VH discriminative accuracy of the clock drawing was comparable to that of the more elaborate test of visual perception, the HVOT. Conclusions: In DLB impaired drawing and visual organization, but not copying ability is associated with the presence of VH. The simple clock drawing test can be helpful to alert a clinician to the possibility of VH in DLB. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Light microscopic hair shaft analysis in ectodermal dysplasia syndromes.
The objective of the study was to catalog hair shaft abnormalities in individuals with ectodermal dysplasia (ED) syndromes using light microscopy and to compare findings with those in unaffected controls. Light microscopy was performed in a nonblinded manner on hair shafts from 65 participants with seven types of ED (hypohidrotic ED, ED-ectrodactyly-cleft lip or palate, ankyloblepharon-ectodermal defects-cleft lip and palate, Clouston syndrome, Goltz syndrome, Schopf-Schulz Passarge syndrome, and oculodentodigital dysplasia) and 41 unaffected controls. Hair donations were collected at the 28th Annual National Family Conference held by the National Foundation for Ectodermal Dysplasia. Control participants were recruited from a private dermatology practice and an academic children's hospital outpatient dermatology clinic. Sixty-five affected participants and 41 unaffected controls were included in the analysis. We assessed the hair shafts of ED and control participants for abnormalities visible using LM. Light microscopy identified various pathologic hair shaft abnormalities in each type of ED, although none of the findings were statistically significantly different from those of the control group. Light microscopy is a poor adjuvant tool in the diagnosis of ED syndromes. Most findings are nonspecific and not sufficiently sensitive.


Entamoeba histolytica causes intestinal secretion: role of serotonin.
Lysates of the protozoan parasite Entamoeba histolytica altered active electrolyte transport when present on the serosal surface of rabbit ileum and rat colon. The lysate-induced effects on electrolyte transport were similar to those caused by serotonin, and were blocked by bufotenine, an analog known to inhibit the action of serotonin. The transport effects were partially inhibited by antibody to serotonin. The amebic lysates were shown to contain serotonin by radioimmunoassay, high-performance liquid chromatography, and thin-layer chromatography. These results suggest that the serotonin present in Entamoeba histolytica may be important in the diarrhea seen in amebiasis.


Rocks, skulls and materialism: geology and phrenology in late-Georgian Belfast.
Recent years have seen the development of a more nuanced understanding of the emergence of scientific naturalism in the nineteenth century. It has become apparent that scientific naturalism did not emerge sui generis in the years following the publication of Charles Darwin's On the origin of species (1859), but was present, if only in incipient form, much earlier in the century. Building on recent scholarship, this article adopts a geographically focused approach and explores debates about geology and phrenology-two of the diverse forms of knowledge that contributed to scientific naturalism-in late-Georgian Belfast. Having provided the venue for John Tyndall's infamous 1874 address as president of the British Association for the Advancement of Science, Belfast occupies a central place in the story of nineteenth-century scientific naturalism. However, in uncovering the intricate and surprising ways in which scientific knowledge gained, or was denied, epistemic and civic credibility in Belfast, this discussion will demonstrate that naturalism, materialism and the relationship between science and religion were matters of public debate in the town long before Tyndall's intervention.


Two-dimensional mapping of underdosed areas using radiochromic film for patients undergoing total skin electron beam radiotherapy.
To demonstrate the viability of radiochromic film as an in vivo, two-dimensional dosimeter for the measurement of underdosed areas in patients undergoing total skin electron beam (TSEB) radiotherapy. The results were compared with thermoluminescent dosimeter measurements. Dosimetry results are reported for an inframammary fold of 2 patients treated using a modified version of the Stanford six-position (i.e., six-field and dual-beam) TSEB technique. The results are presented as contour plots of film optical density and percentage of dose. A linear dose profile measured from film was compared with the thermoluminescent dosimeter measurements. The results showed that the percentage doses as measured by film are in good agreement with those measured by the thermoluminescent dosimeters. The isodose contour plots provided by film can be used as a two-dimensional dose map for a patient when determining the size of the supplemental patch fields. Radiochromic film is a viable dosimetry tool that the radiation oncologist can use to understand the surface dose heterogeneity better across complex concave regions of skin to help establish more appropriate margins to patch underdosed areas. Film could be used for patients undergoing TSEB for disorders such as mycosis fungoides or undergoing TSEB or regional skin electron beam for widespread skin metastases from breast cancer and other malignancies.


Cellular strategies for enhancement of fracture repair.
Tissue engineering seeks to translate scientific knowledge into tangible products to advance the repair, replacement, or regeneration of organs and tissues. Current tissue engineering strategies have progressed recently from a historical approach that is based primarily on biomaterials to a cell and tissue-based approach that includes understanding of cell-sourcing and bioactive stimuli. New options include methods for harvest and transplantation of tissue-forming cells, bioactive matrix materials that act as tissue scaffolds, and delivery of bioactive molecules within scaffolds. These strategies are already benefiting patients, and they place increasing demands on orthopaedic surgeons to have a solid foundation in the contemporary concepts and principles of cell-based tissue engineering. Essentially all orthopaedic tissue engineering strategies can be distilled to a strategy or combination of strategies that seek to increase the number or relative performance of bone-forming cells. The global term connective tissue progenitors has been used to define the heterogeneous populations of stem and progenitor cells that are found in native tissue and that are capable of differentiating into one or more connective tissue phenotypes. These stem or progenitor populations are found in various tissue sources, with varying degrees of ability to differentiate along connective tissue lineages. Available cell-based strategies include targeting local cells with use of scaffolds or bioactive factors, or transplantation of autogenous connective tissue progenitor cells derived from bone marrow or other tissues, with or without processing to change their concentration or prevalence. The future may include means of homing circulating connective tissue progenitor cells with use of intrinsic chemokine systems, or modifying the biological performance of connective tissue progenitor cells by means of genetic modifications.


Self-expandable metal stents for obstructing colonic and extracolonic cancer: European Society of Gastrointestinal Endoscopy (ESGE) Guideline - Update 2020.
The following recommendations should only be applied after a thorough diagnostic evaluation including a contrast-enhanced computed tomography (CT) scan. 1 : ESGE recommends colonic stenting to be reserved for patients with clinical symptoms and radiological signs of malignant large-bowel obstruction, without signs of perforation. ESGE does not recommend prophylactic stent placement.Strong recommendation, low quality evidence. 2 : ESGE recommends stenting as a bridge to surgery to be discussed, within a shared decision-making process, as a treatment option in patients with potentially curable left-sided obstructing colon cancer as an alternative to emergency resection.Strong recommendation, high quality evidence. 3 : ESGE recommends colonic stenting as the preferred treatment for palliation of malignant colonic obstruction.Strong recommendation, high quality evidence. 4 : ESGE suggests consideration of colonic stenting for malignant obstruction of the proximal colon either as a bridge to surgery or in a palliative setting.Weak recommendation, low quality evidence. 5 : ESGE suggests a time interval of approximately 2 weeks until resection when colonic stenting is performed as a bridge to elective surgery in patients with curable left-sided colon cancer.Weak recommendation, low quality evidence. 6 : ESGE recommends that colonic stenting should be performed or directly supervised by an operator who can demonstrate competence in both colonoscopy and fluoroscopic techniques and who performs colonic stenting on a regular basis.Strong recommendation, low quality evidence. 7 : ESGE suggests that a decompressing stoma as a bridge to elective surgery is a valid option if the patient is not a candidate for colonic stenting or when stenting expertise is not available.Weak recommendation, low quality evidence.


Differential actions of pacific ciguatoxin-1 on sodium channel subtypes in mammalian sensory neurons.
Pacific ciguatoxin-1 (P-CTX-1), is a highly lipophilic cyclic polyether molecule originating from the marine dinoflagellate Gambierdiscus toxicus. Its effects were investigated on sodium channel subtypes present in acutely dissociated rat dorsal root ganglion neurons, using whole-cell patch clamp techniques. Concentrations of P-CTX-1 ranging from 0.2 to 20 nM had no effect on the kinetics of tetrodotoxin-sensitive (TTX-S) or tetrodotoxin-resistant (TTX-R) sodium channel activation and inactivation, however, a concentration-dependent reduction in peak current amplitude occurred in both channel types. The main actions of 5 nM P-CTX-1 on TTX-S sodium channels were a 13-mV hyperpolarizing shift in the voltage dependence of sodium channel activation and a 22-mV hyperpolarizing shift in steady-state inactivation (hinfinity). In addition, P-CTX-1 caused a rapid rise in the membrane leakage current in cells expressing TTX-S sodium channels. This effect was blocked by 200 nM TTX, indicating an action mediated through TTX-S sodium channels. In contrast, the main action of P-CTX-1 (5 nM) on TTX-R sodium channels was a significant increase in the rate of recovery from sodium channel inactivation. These results indicate that P-CTX-1 acts to modify voltage-gated sodium channels present in peripheral sensory neurons consistent with its action to increase nerve excitability. This provides an explanation for the sensory neurological disturbances associated with ciguatera fish poisoning.


Seasonal and weekly patterns of hospital admissions for acute diverticulitis.
Onset and hospitalization of acute diseases do not occur randomly, but exhibit preferred high-risk temporal periods. The aim of this study, based on the database of hospital admissions of the Emilia-Romagna region of Italy, was to evaluate the possible existence of a seasonal or weekly pattern of hospitalization for acute diverticulitis (AD), and different rates of complications between weekend (WE) vs. weekday (WD) admissions. The study included all emergency hospital admissions in Emilia Romagna Region for AD between 1999 and 2011 (ICD-9-CM codes: 562.11-562.13). Day of admission was categorized, respectively, into four 3-month intervals, twelve 1-month intervals, seven 1-day intervals for statistical analysis, performed by c2 test goodness of fit and partial Fourier series on total number of cases, males and females, nonfatal or fatal cases, without and with hemorrhage. The database contained records of 29,428 events of AD, relative to 24,843 different patients (mean age: 71.2 ± 13.8 years; 40.5% males). Chronobiological analysis yielded a biphasic rhythmic pattern in AD admissions, characterized by two peaks in Autumn and Spring. As for day of admission, a progressive decrease of frequency during the week was observed. In turn, a slight increase of admissions on WE was observed for hemorrhagic events. An excess burden of hospitalization for AD is observed in the region Emilia-Romagna of Italy, with demonstration of a biphasic cyclical pattern with peaks in Autumn and Spring. Again, a decreasing number of Monday to Friday admissions was observed. Further studies are needed to identify possible underlying causes.


Magnitude of Hematoma Volume Measurement Error in Intracerebral Hemorrhage.
Limiting intracerebral hemorrhage (ICH) and intraventricular hemorrhage (IVH) expansion is a common target for acute ICH studies and, therefore, accurate measurement of hematoma volumes is required. We investigated the amount of hematoma volume difference between computed tomography scans that can be considered as measurement error. Five raters performed baseline (<6 hours) and 24-hour total hematoma (ICH+IVH) computer-assisted volumetric analysis from 40 selected ICH patients from the Predicting Hematoma Growth and Outcome in Intracerebral Hemorrhage Using Contrast Bolus CT (PREDICT) study cohort twice. Estimates of intrarater and interrater reliability are expressed as intraclass correlation coefficients and minimum detectable difference (MDD). Total hematoma volumetric analyses had excellent intra- and interrater agreements (intraclass correlation coefficients 0.994 and 0.992, respectively). MDD for intra- and interrater volumes was 6.68 and 7.72 mL, respectively, and were higher the larger total hematoma volume was and in patients with subarachnoid hemorrhage or IVH. MDD for total hematoma volume measurement of 10.4 mL was found in patients with largest hematoma volumes. In patients with subarachnoid hemorrhage or IVH, MDD for total hematoma volume was 10.3 and 10.4 mL, respectively. In patients without IVH, MDD for intra- and interrater pure ICH volumes were 3.82 and 5.83 mL, respectively. A threshold higher than 10.4 mL seems to be reliable to avoid error of total hematoma volume measurement in a broad range of patients. An absolute ICH volume increase of >6 mL, commonly used as outcome in ICH studies, seems well above MDD and, therefore, could be used to reliably detect ICH expansion.


General Practitioners' recommendations of self-directed-exercises for musculoskeletal problems and perceived barriers and facilitators to doing so: a mixed methods study.
Musculoskeletal problems substantially impact the demand for and the finances of the United Kingdom's National Health Service. Some of this demand and cost could be alleviated if patients use self-directed-exercises. The present study aims first to establish whether general practitioners already recommend self-directed-exercises and second to describe barriers and facilitators to making such recommendations. The design of the current study included surveys and interviews. The surveys were designed to draw out participants' tendency to recommend self-directed-exercises and their behavioral drivers to do so. The drivers investigated include 14 domains described by the Theoretical Domains Framework. The surveys were completed online and the responses were analyzed using descriptive reports and regression analyses. The interviews were designed to more fully understand participants' experiences recommending self-directed-exercises according to the same framework. The interviews were audio-taped, transcribed, and thematically analyzed. The survey found that the following domains significantly predicted participants' tendency to recommend self-directed-exercises: Environmental contexts and resources, Goals, Intentions, Knowledge, Memory attention and decision processes, and Social/professional role. The interviews brought out four themes that could be leveraged to increase general practitioners' tendency to recommend self-directed-exercises: (1) Practitioners' beliefs about self-directed-exercises being effective, (2) Patients' motivations to engage in self-directed-exercises, (3) Time constraints, and (4) The ease with which practitioners can recommend self-directed-exercises. Most general practitioners already recommend self-directed-exercises, though they note significant barriers that may prevent them from doing so. General practitioners' tendency to recommend self-directed-exercises would be bolstered by creating a respected central resource of exercise pamphlets. These pamphlets should clearly describe how different self-directed-exercises should be performed and evidence supporting their effectiveness.


Associations between biological ageing and the risk of, genetic susceptibility to, and life expectancy associated with rheumatoid arthritis: a secondary analysis of two observational studies.
Rheumatoid arthritis is a chronic autoimmune disorder that affects life expectancy. Accelerated biological ageing is thought to be a major risk factor for age-related diseases, but its role in rheumatoid arthritis remains uncertain. We aimed to assess the associations between biological ageing and risk of rheumatoid arthritis and genetic susceptibility to the disease. We also aimed to assess the effect of biological ageing on the life expectancy of people with rheumatoid arthritis. We calculated the chronological age-adjusted biological age-by both the Klemera-Doubal method (KDMAge) and phenotypic age (PhenoAge)-as a surrogate measure for biological ageing in participants from the US National Health and Nutrition Examination Survey (NHANES) and UK Biobank study. KDMAge or PhenoAge acceleration was defined as the residual of the regression of KDMAge or PhenoAge based on chronological age. Participants with accelerated biological ageing had KDMAge or PhenoAge acceleration values greater than 0, whereas those without accelerated ageing had values less than or equal to 0. We did cross-sectional analyses to assess the association between biological ageing and prevalent rheumatoid arthritis in both cohorts and prospective analyses to assess the association between biological ageing and incident rheumatoid arthritis in the UK Biobank. Logistic regression and Cox proportional hazards models were used to analyse these associations. Polygenic risk scores were used to establish genetic susceptibility to rheumatoid arthritis and to analyse the interaction between biological ageing and genetic risk. We also assessed the association between life expectancy and biological ageing status in people with rheumatoid arthritis. In the cross-sectional analyses, each 1-year increase in age-adjusted biological age was associated with an increase in the risk of rheumatoid arthritis of between 1% and 10%. In the NHANES, individuals with accelerated ageing had a higher risk of rheumatoid arthritis than non-accelerated ageing individuals, with odds ratios of 1·21 (95% CI 1·03-1·42; p=0·018) for KDMAge acceleration and 1·46 (1·26-1·69; p<0·0001) for PhenoAge acceleration. Similarly, in the UK Biobank, the risk of rheumatoid arthritis was increased in individuals with accelerated ageing compared with individuals with no accelerated ageing (KDMAge odds ratio 1·96 [95% CI 1·71-2·24]; PhenoAge 2·71 [2·51-2·92]). In the prospective analyses of the UK Biobank population, accelerated biological ageing was associated with an increased risk of incident rheumatoid arthritis as measured by both KDMAge (hazard ratio 1·27 [95% CI 1·03-1·55]) and PhenoAge (1·70 [1·52-1·92]). Among participants with high genetic predisposition to rheumatoid arthritis, accelerated biological ageing was associated with an increased risk of incident disease, and we noted significant additive interactions between accelerated biological ageing and genetic risk. At age 45 years, people with rheumatoid arthritis had reduced life expectancy compared with those without rheumatoid arthritis. Among people with rheumatoid arthritis, accelerated biological ageing was associated with reduced life expectancy compared with not having accelerated biological ageing. Accelerated biological ageing could increase the risk of rheumatoid arthritis, especially among people with high genetic risk, and could reduce the life expectancy of people with rheumatoid arthritis. The identification of populations with accelerated biological ageing has important implications for reducing the risk of rheumatoid arthritis and of lowered life expectancy. National Natural Science Foundation of China.


[Epidemiology of bone and joint disease - the present and future - . Epidemiology of falls].
A longitudinal analysis using a population-based cohort study known as Research on Osteoarthritis/osteoporosis Against Disability (ROAD) clarified the associations of physical performance and bone and joint diseases with falls. A total of 452 men and 896 women were analyzed in the present study (mean age, 63.9 years). A questionnaire was used to assess the number of falls during the 3-year follow-up. Grip strength, 6-m walking time, and chair stand time were measured at baseline. Knee osteoarthritis (OA) and lumbar spondylosis were defined as Kellgren Lawrence = 3 or 4. Vertebral fracture (VFx) was assessed with the Japanese Society of Bone and Mineral Research criteria. Knee and lower back pain were estimated by an interview. During a 3-year follow-up, 17.4% men and 24.1% women reported at least one fall. Multinomial logistic regression analysis after adjustment for age and BMI showed that a longer 6-m walking time was a risk factor for multiple falls in men and women. Knee pan was also a risk factor for multiple falls in women. Walking time and knee pain may be a simple and quick option for measuring the risk of falling.


Deep subwavelength lithography via tunable terahertz plasmons.
A scheme to overcome diffraction limit in optical lithography via tunable plasmons is proposed. The plasmons are generated by a current-driven instability and are resonance amplified between the drain and source barriers of the transistor. A series of discrete deep subwavelength can be obtained by controlling the gate voltage. Thus, it is possible to realize lithography with a resolution over 1/100 vacuum wavelength and achieve arbitrary one-dimensional and even simple two-dimensional patterns. Our scheme works in the linear optics regime and is easy to be experimentally realized.


The anti-inflammatory effect of triphala in arthritic-induced rats.
Triphala, an Indian Ayurvedic herbal formulation which contains Terminalia chebula Retz. (Combretaceae), Terminalia bellerica (Gaertn.) Roxb. (Combretaceae) and Emblica officinalis L. (Phyllanthaceae), is used for treating bowel-related complications, inflammatory disorders, and gastritis. To determine the anti-arthritic effect of triphala in arthritis-induced rats. For comparison purpose, the non-steroidal anti-inflammatory drug indomethacin was used. Arthritis was induced in Wistar albino rats by intradermal injection of complete Freund's adjuvant (0.1 ml) into the foot pad of right hind paw. Triphala (100 mg/kg b wt, i.p.) was administered from day 11 to 18 after the administration of complete Freund's adjuvant. The activities/levels of lysosomal enzymes, glycoproteins, antioxidant status, and lipid peroxidation were determined in the paw tissues of arthritic rats. In addition, the inflammatory mediators were also measured in both the serum and the paw tissue of arthritic rats. The levels/activities of lipid peroxidation (∼41.5%), glycoproteins (hexose ∼43.3%, hexosamine ∼36.5%, and sialic acid ∼33.7%), lysosomal enzymes (acid phosphatase ∼52.4%, β-galactosidase ∼22.9%, N-acetyl β-glucosaminidase ∼22.1%, and cathepsin-D ∼27.7%) were found to be decreased and the antioxidant status (SOD ∼75.6%, CAT ∼62.7%, GPx ∼55.8%, GST ∼82.1%, and GSH ∼72.7%) was increased in the paw tissues of triphala-treated arthritic rats. In addition, the inflammatory mediator levels in serum (TNF-α ∼75.5%, IL-1β ∼99%, VEGF ∼75.2%, MCP-1 ∼76.4%, and PGE2 ∼69.9%) and in paw tissues (TNF-α ∼71.6%, IL-1β ∼75.5%, VEGF ∼55.1%, MCP-1 ∼69.1%, and PGE2 ∼66.8%) were found to be suppressed. Triphala has a promising anti-inflammatory effect in the inflamed paw of arthritis-induced rats.


Phosphoinositide 3-kinase forms a complex with platelet membrane glycoprotein Ib-IX-V complex and 14-3-3zeta.
The binding of von Willebrand factor (vWF) to glycoprotein (GP) Ib-IX-V stimulates transmembrane signaling events that lead to platelet adhesion and aggregation. Recent studies have revealed that the signaling protein 14-3-3 zeta binds directly to the cytoplasmic domain of GP Ib alpha. In this study, the dynamic association of 14-3-3 zeta with GP Ib-IX, the phosphoinositide 3-kinase (PI 3-kinase), or both, was investigated in resting, thrombin, or vWF and botrocetin-stimulated platelets by analysis of discrete subcellular fractions. Results of this study demonstrate maximal coimmunoprecipitation of 14-3-3 zeta with GP Ib-IX in the nonstimulated cytosolic fraction and in the actin cytoskeletal fraction of thrombin- or vWF-stimulated human platelets. Immunoprecipitated 14-3-3 zeta or GP Ib from cytosolic fractions contained PI 3-kinase enzyme activity and an 85-kd polypeptide recognized by antibodies to the p85 subunit of PI 3-kinase. After platelet activation, the level of association between these species decreased in the cytosolic fraction. However, increased complex formation between 14-3-3 zeta and GP Ib-IX and between PI 3-kinase and GP Ib-IX was detected in actin cytoskeletal fractions derived from thrombin- or vWF-stimulated platelets. Recombinant glutathione S-transferase-14-3-3 zeta fusion protein (14-3-3 zeta-GST) inhibited affinity-captured PI 3-kinase enzyme activity up to 70% at 2 mcmol/L 14-3-3 zeta-GST. However, increasing concentrations up to 5 mcmol/L 14-3-3 zeta-GST resulted in the 3-fold enhancement of PI 3-kinase enzyme activity. We propose that the association between PI 3-kinase and 14-3-3 zeta with GP Ib-IX serves to promote the rapid translocation of these signaling proteins to the activated cytoskeleton, thereby regulating the formation of 3-position phosphoinositide-signaling molecules in this subcellular compartment. (Blood. 2000;96:577-584)


3,5-Di-O-caffeoyl-epi-quinic acid from the leaves and stems of Erigeron annuus inhibits protein glycation, aldose reductase, and cataractogenesis.
In our ongoing project directed toward the discovery of novel treatments for diabetic complications from herbal medicines, sixteen compounds including three caffeoylquinic acids and four flavonoids were isolated from an EtOAc-soluble extract of the stems and leaves of Erigeron annuus. All the isolates were evaluated in vitro for inhibitory activity on the formation of advanced glycation end products (AGEs) and rat lens aldose reductase (RLAR). Of these, 3,5-di-O-caffeoyl-epi-quinic acid (3) exhibited the most potent inhibitory activity in both the AGEs and aldose reductase (AR) assays. Compound 3 markedly reduced AGEs-bovin serum albumin (BSA) cross-linking in a dose-dependent manner. Furthermore, opacity of lenses was significantly prevented when they were treated with 3 in an ex vivo experiment.


Multiplex analysis of enzyme kinetics and inhibition by droplet microfluidics using picoinjectors.
Enzyme kinetics and inhibition is important for a wide range of disciplines including pharmacology, medicine and industrial bioprocess technology. We present a novel microdroplet-based device for extensive characterization of the reaction kinetics of enzyme substrate inhibitor systems in a single experiment utilizing an integrated droplet picoinjector for bioanalysis. This device enables the scanning of multiple fluorescently-barcoded inhibitor concentrations and substrate conditions in a single, highly time-resolved experiment yielding the Michaelis constant (Km), the turnover number (kcat) and the enzyme inhibitor dissociation constants (ki, ki'). Using this device we determine Km and kcat for β-galactosidase and the fluorogenic substrate Resorufin β-D-galactopyranoside (RBG) to be 442 μM and 1070 s(-1), respectively. Furthermore, we examine the inhibitory effects of isopropyl-β-D-thiogalactopyranoside (IPTG) on β-galactosidase. This system has a number of potential applications, for example it could be used to screen inhibitors to pharmaceutically relevant enzymes and to characterize engineered enzyme variants for biofuels production, in both cases acquiring detailed information about the enzyme catalysis and enzyme inhibitor interaction at high throughput and low cost.


Pathology Reporting in Neuroendocrine Neoplasms of the Digestive System: Everything You Always Wanted to Know but Were Too Afraid to Ask.
During the 5th NIKE (Neuroendocrine tumors Innovation in Knowledge and Education) meeting, held in Naples, Italy, in May 2019, discussions centered on the understanding of pathology reports of gastroenetropancreactic neuroendocrine neoplasms. In particular, the main problem concerned the difficulty that clinicians experience in extrapolating relevant information from neuroendocrine tumor pathology reports. During the meeting, participants were asked to identify and rate issues which they have encountered, for which the input of an expert pathologist would have been appreciated. This article is a collection of the most rated questions and relative answers, focusing on three main topics: 1) morphology and classification; 2) Ki67 and grading; 3) immunohistochemistry. Patient management should be based on multidisciplinary decisions, taking into account clinical and pathology-related features with clear comprehension between all health care professionals. Indeed, pathologists require clinical details and laboratory findings when relevant, while clinicians require concise and standardized reports. In keeping with this last statement, the minimum requirements in pathology datasets are provided in this paper and should be a baseline for all neuroendocrine tumor professionals.


Salmonella-associated conjunctivitis in a cat.
Salmonella typhimurium-associated conjunctivitis in an adult female cat was characterized by epiphora and blepharospasm. The cat also harbored intestinal S typhimurium, as did one other cat in the same shipment of 7 cats obtained from a commercial cat dealer. The 3 isolates were resistant to ampicillin, cephaloridine, cephalothin, streptomycin, triple sulfonamides, and tetracycline. All 3 isolates were capable of transferring a part of the antibiotic resistance pattern to an Escherichia coli K-12 recipient. Salmonella were not recovered from the conjunctiva or fecal flora of either cat after 10 days of therapy with chloramphenicol.


MRI Radiomics for the Prediction of Fuhrman Grade in Clear Cell Renal Cell Carcinoma: a Machine Learning Exploratory Study.
The Fuhrman nuclear grade is a recognized prognostic factor for patients with clear cell renal cell carcinoma (CCRCC) and its pre-treatment evaluation significantly affects decision-making in terms of management. In this study, we aimed to assess the feasibility of a combined approach of radiomics and machine learning based on MR images for a non-invasive prediction of Fuhrman grade, specifically differentiation of high- from low-grade tumor and grade assessment. Images acquired on a 3-Tesla scanner (T2-weighted and post-contrast) from 32 patients (20 with low-grade and 12 with high-grade tumor) were annotated to generate volumes of interest enclosing CCRCC lesions. After image resampling, normalization, and filtering, 2438 features were extracted. A two-step feature reduction process was used to between 1 and 7 features depending on the algorithm employed. A J48 decision tree alone and in combination with ensemble learning methods were used. In the differentiation between high- and low-grade tumors, all the ensemble methods achieved an accuracy greater than 90%. On the other end, the best results in terms of accuracy (84.4%) in the assessment of tumor grade were achieved by the random forest. These evidences support the hypothesis that a combined radiomic and machine learning approach based on MR images could represent a feasible tool for the prediction of Fuhrman grade in patients affected by CCRCC.


[Croatian Medical Association--Branch Zagreb].
The available literature shows us that "Druztvo ljeciteljah u Zagrebus (the Society of Healers in Zagreb) was founded as far back as the year 1845 by a total of thirteen members. This data allows us to follow the role of doctors and health workers in Zagreb through their everyday profession, research, organizational and social work as well as management through a period of over one hundred to seventy years. The Branch Zagreb was active before the official establishment of subsidiaries of CMA which is evident from the minutes of the regular annual assembly of the Croatian Medical Association on 21 March 1948. Until the end of 1956, there was no clear division of labor, functions and competencies between the Branch and the Main Board. Their actions were instead consolidated and the Branch operated within and under the name of Croatian Medical Association. In that year the Branch became independent. The Branch Zagreb is the largest and one of the most active branches of the Croatian Medical Association. At the moment, the Branch brings together 3621 members, regular members--doctors of medicine (2497), doctors of dental medicine (384), retired physicians (710), and associate members (30 specialists with higher education who are not doctors). The Branch is especially accomplished in its activities in the area of professional development of its members and therefore organizes a series of scientific conferences in the framework of continuous education of physicians, allowing its members to acquire necessary points for the extension of their operating license. The choir "Zagrebacki lijecnici pjevaci" (Zagreb Physicians' Choir) of the Croatian Medical Music Society of the CMA and its activities are inseparable from the Branch Zagreb. The Branch is firmly linked to the parent body, the CMA, and thus has a visible impact on the strategy and the activities of the Association as a whole. Most professional societies of the CMA have their headquarters in Zagreb and this is inevitably reflected in their overall function. The Branch Zagreb supports the harmonious development of health care throughout the territory of the Republic of Croatia. Cities of Zagreb and Mainz, are cities--friends and the CMA Branch Zagreb works together and exchanges visits with mem- bers of the Medical Society of Mainz for years. Next year professional and scientific cooperation between the Branch Za- greb of CMA and the Medical Society of Mainz will mark four decades.


Mares prefer the voices of highly fertile stallions.
We investigated the possibility that stallion whinnies, known to encode caller size, also encoded information about caller arousal and fertility, and the reactions of mares in relation to type of voice. Voice acoustic features are correlated with arousal and reproduction success, the lower-pitched the stallion's voice, the slower his heart beat and the higher his fertility. Females from three study groups preferred playbacks of low-pitched voices. Hence, females are attracted by frequencies encoding for large male size, calmness and high fertility. More work is needed to explore the relative importance of morpho-physiological features. Assortative mating may be involved as large females preferred voices of larger stallions. Our study contributes to basic and applied ongoing research on mammal reproduction, and questions the mechanisms used by females to detect males' fertility.


Cooperation and Competition with Hyperscanning Methods: Review and Future Application to Emotion Domain.
Cooperation and competition, as two common and opposite examples of interpersonal dynamics, are thought to be reflected by different cognitive, neural, and behavioral patterns. According to the conventional approach, they have been explored by measuring subjects' reactions during individual performance or turn-based interactions in artificial settings, that don't allow on-line, ecological enactment of real-life social exchange. Considering the importance of these factors, and accounting for the complexity of such phenomena, the hyperscanning approach emerged as a multi-subject paradigm since it allows the simultaneous recording of the brain activity from multiple participants interacting. In this view, the present paper aimed at reviewing the most significant work about cooperation and competition by EEG hyperscanning technique, which proved to be a promising tool in capturing the sudden course of social interactions. In detail, the review will consider and group different experimental tasks that have been developed so far: (1) paradigms that used rhythm, music and motor synchronization; (2) card tasks taken from the Game Theory; (3) computerized tasks; and (4) possible real-life applications. Finally, although highlighting the potential contribution of such approach, some important limitations about these paradigms will be elucidated, with a specific focus on the emotional domain.


Fine characterization of the recurrent c.1584+18672A>G deep-intronic mutation in the cystic fibrosis transmembrane conductance regulator gene.
Splicing mutations account for approximately 12% of the 1,890 cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations described in cystic fibrosis (CF). However, their impact on pre-mRNA processing frequently remains unclear. An interesting opportunity to study CFTR transcripts in vivo involves the use of RNA from nasal brushings. Through this approach we previously identified a deep-intronic mutation (c.1584+18672A>G) that activates a 104-base pair (bp) out-of-frame pseudoexon by creating a donor splice site. The screening of 230 patients with CF identified c.1584+18672A>G in three additional individuals, demonstrating that it is a recurrent, and potentially overlooked, mutation among Italian patients. Haplotype analysis suggests that it originated from at least two independent events. To characterize the mutation further, a genomic region, including the activated pseudoexon and surrounding intronic sequences, was cloned into an expression vector and transfected into HeLa cells. RT-PCR analysis identified two alternative splicing products, produced by the activation of two different cryptic acceptor splice sites. One included the 104-bp pseudoexon (78.7% of transcripts), and the other led to the inclusion of a 65-bp pseudoexon (21.3% of mRNAs). The allele-specific measurement of wild-type and aberrant splicings from the nasal-brushing RNA of the three probands with genotype F508del/c.1584+18672A>G demonstrated: (1) a low level of pseudoexon inclusion in the F508del transcript (not containing the splicing mutation); (2) residual wild-type splicing in the c.1584+18672A>G mRNA; (3) the degradation of aberrant transcripts; and (4) the relative strength of the different cryptic splice sites. Interestingly, the residual wild-type splicing detected in transcripts bearing the c.1584+18672A>G mutation correlates well with the milder clinical phenotype of patients.


Medical assessment of the patient undergoing revision total hip arthroplasty.
As the geriatric population in the United States continues to increase relative to the general population, total joint arthroplasty has become an accepted medical tool in our medical armamentarium. Much of the perioperative milieu of the geriatric population tends to be poorly researched and minimally reported within the medical community. An enhanced appreciation of predictable preoperative and postoperative clinical considerations should optimize the availability of this technology and minimize morbidity and mortality.


Deciphering the oligodendrogenic program of neural progenitors: cell intrinsic and extrinsic regulators.
In the developing and adult CNS, neural stem/progenitor cells (NSPCs) and oligodendroglial progenitor cells (OPCs) follow an oligodendrogenic process with the aim of myelinating axons. This process is to a high degree regulated by an oligodendrogenic program (OPr) composed of intrinsic and extrinsic factors that modulate the different steps required for NSPCs to differentiate into myelinating oligodendrocytes. Even though NSPCs and OPCs are present in the diseased CNS and have the capacity to generate oligodendrocytes, sparse remyelination of axons constitutes a major constraint in therapies toward multiple sclerosis (MS) and spinal cord injury (SCI). Lack of pro-oligodendrogenic factors and presence of anti-oligodendrogenic activities are thought to be the main reasons for this limitation. Thus, molecular and cellular strategies aiming at remyelination and at targeting such pro- and anti-oligodendrogenic mechanisms are currently under investigation. The present review summarizes the current knowledge on the OPr; it implements our own findings on mesenchymal stem cell-derived pro-oligodendroglial factors and on the role of p57/kip2 in oligodendroglial differentiation. Moreover, it describes molecular and cellular approaches for the development of future therapies toward remyelination.


Intravenous Tranexamic Acid Decreases Allogeneic Transfusion Requirements in Periacetabular Osteotomy.
Bernese (Ganz) periacetabular osteotomy is associated with significant blood loss and the need for perioperative transfusion. Tranexamic acid decreases blood loss and minimizes transfusion rates in total joint arthroplasty. However, no reports have described its use in patients undergoing Bernese periacetabular osteotomy. This study reports the use of intravenous tranexamic acid in these patients. The study included 137 patients (150 hips) who underwent isolated periacetabular osteotomy at a single institution between 2003 and 2014. Of these, 68 patients (75 hips) received intravenous tranexamic acid 1 g at the time of incision and 1 g at the time of closure. A group of 69 patients (75 hips) served as control subjects who underwent periacetabular osteotomy without administration of intravenous tranexamic acid. Thromboembolic disease was defined as deep venous thrombosis or pulmonary embolism occurring within 6 weeks of surgery. Outcomes measured included transfusion requirements, pre- and postoperative hemoglobin values, operative times, and thromboembolic disease rates. Aspirin was used as the thromboembolic prophylactic regimen in 95% of patients. The rate of allogeneic transfusion was 0 in the tranexamic acid group compared with 21% in the control group (P=.0001). No significant difference was found in the autologous cell salvage requirement (.96 vs 1.01; P=.43) or the thromboembolic disease rate between the tranexamic acid group and the control group (2.67% vs 1.33%; P=.31). The use of intravenous tranexamic acid led to a decreased transfusion requirement with no increased risk of thromboembolic disease in this contemporary cohort of patients undergoing periacetabular osteotomy.


Relationship between maternal periodontal disease and Apgar score of newborns.
The aim of this study was to evaluate the relationship between maternal periodontal disease and the health status of newborns using Apgar scores. One hundred pregnant women with periodontal disease were included in the case series and 100 pregnant women without periodontal disease were placed in the control group, respectively. The periodontal parameters of bleeding on probing (BOP), clinical attachment loss (CAL), probing depth (PD), birth weight, and Apgar scores were recorded in both groups. T-tests and Pearson's correlation coefficient were used to determine the birth weight odds ratio to analyze the relationship between the periodontal parameters of BOP, CAL, and PD on the one hand and an Apgar score of less than 7. An unpaired Student's t-test was used to analyze differences in means between the case and control groups using SPSS ver. 13. The means of the ages, periodontal pocket depths, attachment loss, areas with BOP, Apgar score in the first 5 minutes and infant birth weight exhibited statistically significant differences between the case and control groups. The ratio of an Apgar score of <7 to periodontal disease was 3.14; the ratio of low birth weight risk in mothers with periodontal disease to that in mothers without periodontal disease was 2.74. Pearson's correlation coefficient revealed a significant correlation between the infant birth weight and BOP, CAL, and PD of the mother. In addition, there was a significant correlation between the Apgar score and BOP, CAL, and PD and also between the Apgar score and infant birth weight. The results of this study showed a significant relationship between periodontal disease and infant birth weight; in addition, there was a significant relationship between the periodontal indexes of BOP, CAL, and PD on the one hand and the Apgar score on the other.


Drainage of a Subphrenic Abscess Followed by Two-Stage Gastrectomy and Adjuvant Hyperthermic Intraperitoneal Chemotherapy (HIPEC) for Perforated Gastric Carcinoma: A Case Report.
BACKGROUND Perforated gastric cancer accounts for less than 1% of patients who present with an acute abdomen and for up to 16% of all gastric perforations. A two-stage laparoscopic procedure may be the therapeutic strategy of choice in selected patients, and adjuvant hyperthermic intraperitoneal chemotherapy (HIPEC) can reduce the incidence of peritoneal recurrence. A rare case of subphrenic abscess and gastric perforation due to carcinoma of the gastric fundus, followed by two-stage gastrectomy and adjuvant HIPEC is presented. CASE REPORT A 65-year old man presented with a left subphrenic abscess secondary to perforated gastric carcinoma. Laparoscopic drainage of the abscess was performed. Ten days later, following recovery from sepsis, the patient underwent total laparoscopic gastrectomy, and adjuvant HIPEC followed by a Roux-en-Y esophagojejunostomy. Histopathology showed an intestinal-type gastric adenocarcinoma. The tumor was staged as pT4aN0. The postoperative course was uneventful except for transient atrial fibrillation. The patient was discharged home on postoperative day 11. Systemic adjuvant chemotherapy was begun one month later. At six-month follow-up, the patient had no discomfort on eating or any other symptoms. CONCLUSIONS In this case, a two-stage laparoscopic treatment for perforated gastric carcinoma combined with adjuvant HIPEC was feasible and safe and may be considered at the time of laparoscopic gastrectomy in selected patients with perforated gastric carcinoma.


Hypomagnesemia associated with chemotherapy in patients treated for acute lymphoblastic leukemia: possible mechanisms.
Body weight, height and lumbar spine bone mineral density (BMD) were measured in 40 children (27 male, 13 female, aged 0.3-17.0 years) with acute lymphoblastic leukemia (ALL) at diagnosis and 6 months after initiation of chemotherapy, and in 40 age- and sex-matched local healthy children. Serum and urinary biochemical indices of mineral metabolism were measured in the children with ALL at both time points. From diagnosis to 6 months, a reduction in the fractional excretion of magnesium was found. Serum osteocalcin was low at diagnosis and increased during chemotherapy, whereas 24-h urinary type I collagen cross-linked N-telopeptide was unchanged. The Z-scores for lumbar spine BMD increased and were correlated with the serum osteocalcin at 6 months. The change in serum magnesium was correlated negatively with the change in lumbar spine BMD, and with the lumbar spine BMD Z-score at 6 months. After initiation of treatment for ALL, rapid recovery in bone formation, which results in the movement of extracellular magnesium into the skeleton through bone formation, may be an important contributor to hypomagnesemia.


Arachnoidal cap cell hyperplasia of the intracanalicular optic nerve sheath.
Fifty-two intracanalicular optic nerves from 26 consecutive cadavers at the Dade County Medical Examiner's Office were examined for arachnoidal cap cells three cells thick or greater. Twenty specimens (77%) fulfilled these criteria with predominant involvement of the inferior and lateral aspects of the optic nerve sheath. Twenty-four specimens (92%) had psammoma bodies, and 17 specimens (65%) demonstrated corpora amylacea. Psammoma bodies as well as corpora amylacea were found with increasing frequency with age. However, our study did not show that age was an absolute factor toward the development of arachnoidal cap cell hyperplasia. The factors that incite foci of arachnoidal cap cell hyperplasia to greater activity leading in occasional individuals to the development of a meningioma remain to be determined.


In vitro and in silico studies of the inhibitory effects of some novel kojic acid derivatives on tyrosinase enzyme.
Tyrosinase is a key enzyme in pigment synthesis. Overproduction of melanin in parts of the skin results in hyperpigmentation diseases. This enzyme is also responsible for the enzymatic browning in fruits and vegetables. Thus, its inhibitors are of great importance in the medical, cosmetic and agricultural fields. A series of twelve kojic acid derivatives were designed to be evaluated as tyrosinase activity inhibitors. The potential inhibitory activity of these compounds was investigated in silico using molecular docking simulation method. Four compounds with a range of predicted tyrosinase inhibitory activities were prepared and their inhibitory effect on tyrosinase activity was evaluated. The antioxidant properties of these compounds were also investigated by in vitro DPPH (2,2-diphenyl-1-picrylhydrazyl) and hydrogen peroxide scavenging assays. Compound IIId exhibited the highest tyrosinase inhibitory activity with an IC50 value of 0.216 ± 0.009 mM which was in accordance with the in silico ΔGbind results (-13.24 Kcal/mol). Based on the docking studies, from the twelve compounds studied, one (IIId) appeared to have the highest inhibition on tyrosinase activity. This was confirmed by enzyme activity measurements. Compound IIId has an NO2 group which binds to both of Cu(2+) ions located inside the active site of the enzyme. This compound appeared to be even stronger than kojic acid in inhibiting tyrosinase activity. The DPPH free radical scavenging ability of all the studied compounds was more than that of BHT. However, they were not as strong as BHT or gallic acid in scavenging hydrogen peroxide.


Atmospheric Gas-Phase Formation of Methanesulfonic Acid.
Despite its impact on the climate, the mechanism of methanesulfonic acid (MSA) formation in the oxidation of dimethyl sulfide (DMS) remains unclear. The DMS + OH reaction is known to form methanesulfinic acid (MSIA), methane sulfenic acid (MSEA), the methylthio radical (CH3S), and hydroperoxymethyl thioformate (HPMTF). Among them, HPMTF reacts further to form SO2 and OCS, while the other three form the CH3SO2 radical. Based on theoretical calculations, we find that the CH3SO2 radical can add O2 to form CH3S(O)2OO, which can react further to form MSA. The branching ratio is highly temperature sensitive, and the MSA yield increases with decreasing temperature. In warmer regions, SO2 is the dominant product of DMS oxidation, while in colder regions, large amounts of MSA can form. Global modeling indicates that the proposed temperature-sensitive MSA formation mechanism leads to a substantial increase in the simulated global atmospheric MSA formation and burden.


Effects of acute and 2-week administration of oral salbutamol on exercise performance and muscle strength in athletes.
Our objective was to investigate effects of acute and 2-week administration of oral salbutamol on repeated sprint ability, exercise performance, and muscle strength in elite endurance athletes. Twenty male elite athletes [VO2max: 69.4 ± 1.8 (Mean ± SE) mL/min/kg], aged 25.9 ± 1.4 years, were included in a randomized, double-blinded and placebo-controlled parallel study. At baseline, after acute administration, and again after 2-week administration of the study drugs (8 mg salbutamol or placebo), subjects' maximal voluntary contraction (MVC) of m. quadriceps and isometric endurance of m. deltoideus were measured, followed by three repeated Wingate tests. Exercise performance at 110% of VO2max was determined on a bike ergometer. Acute administration of salbutamol increased peak power during first Wingate test by 4.1 ± 1.7% (P < 0.05). Two-week administration of salbutamol increased (P < 0.05) peak power during first and second Wingate test by 6.4 ± 2.0 and 4.2 ± 1.0%. Neither acute nor 2-week administration of salbutamol had any effect on MVC, exercise performance at 110% of VO2max or on isometric endurance. No differences were observed in the placebo group. In conclusion, salbutamol benefits athletes' sprint ability. Thus, the present study supports the restriction of oral salbutamol in competitive sports.


Sperm transport in the reproductive tract of female zebra finches (Taeniopygia guttata).
Aspects of sperm transport in the oviducts of female zebra finches were examined by recording the decline in the number of spermatozoa on the outer perivitelline layer of successively laid eggs. Data from a single clutch of eggs from 32 females indicates that the mean per capita rate of loss of spermatozoa was estimated to be 0.0170 +/- 0.002 SEM spermatozoa h-1. However, individual females showed no consistency in the rate at which spermatozoa were lost from their oviduct over five successive clutches. Models of the mechanism of sperm competition in birds assume that the rate of loss of spermatozoa does not differ between inseminations made before or after the start of egg laying. This assumption was found to be valid: the instantaneous per capita rate of loss of spermatozoa did not differ significantly between females inseminated either before (0.01445 +/- 0.0028 SEM spermatozoa h-1) or after (0.01674 +/- 0.0023 SEM spermatozoa h-1) the onset of oviposition. The rate of sperm transport through the infundibulum was determined to be slower than that between the utero-vaginal sperm storage tubules and the infundibulum by comparing the number of spermatozoa associated with the perivitelline layers of eggs laid after a day on which no eggs were laid. Eggs with < 20 spermatozoa on the outer perivitelline layer were found to have a 50% probability of being infertile. The results are compared with data for domestic fowl and turkeys.


Are long corolla tubes in Pedicularis driven by pollinator selection?
The evolution of long corolla tubes has been hypothesized to be driven by long-tongued pollinators. Corolla tubes in Pedicularis species can be longer than 10 cm which may function as flower stalks to increase visual attractiveness to pollinators because these species provide no nectar and are pollinated by bumblebees. The corolla tube length was manipulated (shorter or longer) in two Pedicularis species in field to examine whether longer tubes are more attractive to pollinators and produce more seeds than short tubes. Our results did not support the pollinator attraction hypothesis, leaving the evolution of long tubes in Pedicularis remains mysterious.


[Psychiatric features of patients with duodenal ulcer at various stages of the disease].
To examine patients with duodenal ulcer (DU) at different stages of the disease for psychic alterations. Psychodiagnostic techniques were employed in examination of 192 patients with DU (81 new cases of the disease, 88 cases of recurrent exacerbations, 23 cases with persistent remission for 2 years and longer). Control group consisted of 73 patients without gastrointestinal complaints. Abnormal psychic adaptation was found in 73.3% patients with DU exacerbation, in 47.8% patients in remission and 41.1% controls. Hypochondriac condition and non-adaptation were diagnosed in new cases of DU and recurrent exacerbations with the same frequency. The former had more frequently mixed attitude to the disease and mixed psychopathological syndromes. Psychic compensation rests on information about DU and its prevention which patients receive from the doctor, so these patients benefit much from communication with the physician. Observation over the patient's behavior during such communication allows the doctor to reveal anxiety and depression. Such patients should be referred to consult psychotherapist or psychiatrist. Psychic non-adaptation accompanying development of DU manifests with different clinical symptoms at different DU stages. Diagnosis of mental condition of DU patients is important for the choice of individual treatment approach.


Classification of non-coding variants with high pathogenic impact.
Whole genome sequencing is increasingly used to diagnose medical conditions of genetic origin. While both coding and non-coding DNA variants contribute to a wide range of diseases, most patients who receive a WGS-based diagnosis today harbour a protein-coding mutation. Functional interpretation and prioritization of non-coding variants represents a persistent challenge, and disease-causing non-coding variants remain largely unidentified. Depending on the disease, WGS fails to identify a candidate variant in 20-80% of patients, severely limiting the usefulness of sequencing for personalised medicine. Here we present FINSURF, a machine-learning approach to predict the functional impact of non-coding variants in regulatory regions. FINSURF outperforms state-of-the-art methods, owing in particular to optimized control variants selection during training. In addition to ranking candidate variants, FINSURF breaks down the score for each variant into contributions from individual annotations, facilitating the evaluation of their functional relevance. We applied FINSURF to a diverse set of 30 diseases with described causative non-coding mutations, and correctly identified the disease-causative non-coding variant within the ten top hits in 22 cases. FINSURF is implemented as an online server to as well as custom browser tracks, and provides a quick and efficient solution to prioritize candidate non-coding variants in realistic clinical settings.


Quantitative assessments of traumatic axonal injury in human brain: concordance of microdialysis and advanced MRI.
Axonal injury is a major contributor to adverse outcomes following brain trauma. However, the extent of axonal injury cannot currently be assessed reliably in living humans. Here, we used two experimental methods with distinct noise sources and limitations in the same cohort of 15 patients with severe traumatic brain injury to assess axonal injury. One hundred kilodalton cut-off microdialysis catheters were implanted at a median time of 17 h (13-29 h) after injury in normal appearing (on computed tomography scan) frontal white matter in all patients, and samples were collected for at least 72 h. Multiple analytes, such as the metabolic markers glucose, lactate, pyruvate, glutamate and tau and amyloid-β proteins, were measured every 1-2 h in the microdialysis samples. Diffusion tensor magnetic resonance imaging scans at 3 T were performed 2-9 weeks after injury in 11 patients. Stability of diffusion tensor imaging findings was verified by repeat scans 1-3 years later in seven patients. An additional four patients were scanned only at 1-3 years after injury. Imaging abnormalities were assessed based on comparisons with five healthy control subjects for each patient, matched by age and sex (32 controls in total). No safety concerns arose during either microdialysis or scanning. We found that acute microdialysis measurements of the axonal cytoskeletal protein tau in the brain extracellular space correlated well with diffusion tensor magnetic resonance imaging-based measurements of reduced brain white matter integrity in the 1-cm radius white matter-masked region near the microdialysis catheter insertion sites. Specifically, we found a significant inverse correlation between microdialysis measured levels of tau 13-36 h after injury and anisotropy reductions in comparison with healthy controls (Spearman's r = -0.64, P = 0.006). Anisotropy reductions near microdialysis catheter insertion sites were highly correlated with reductions in multiple additional white matter regions. We interpret this result to mean that both microdialysis and diffusion tensor magnetic resonance imaging accurately reflect the same pathophysiological process: traumatic axonal injury. This cross-validation increases confidence in both methods for the clinical assessment of axonal injury. However, neither microdialysis nor diffusion tensor magnetic resonance imaging have been validated versus post-mortem histology in humans. Furthermore, future work will be required to determine the prognostic significance of these assessments of traumatic axonal injury when combined with other clinical and radiological measures.