Cellulose nanofiber-titania nanocomposites as potential drug delivery systems for dermal applications.
In this work, new efficient drug delivery systems based on cellulose nanofiber-titania nanocomposites grafted with three different types of model drugs such as diclofenac sodium, penicillamine-D and phosphomycin were successfully synthesized and displayed distinctly different controlled long-term release profiles. Three different methods of medicine introduction were used to show that various interactions between TiO2 and drug molecules could be used to control the kinetics of long-term drug release. All synthesis reactions were carried out in aqueous media. The morphology, chemical structure and properties of the obtained materials were characterized by SEM, TEM and AFM microscopy, nanoparticle tracking analysis, X-ray diffraction, and TGA analysis. According to FT-IR and UV-Vis spectroscopy data, the titania binds to cellulose nanofibers via formation of ester bonds and to drug molecules via formation of surface complexes. The drug release kinetics was studied in vitro for diclofenac sodium and penicillamine-D spectrophotometrically and for phosphomycin using a radio-labeling analysis with 33P-marked ATP as a model phosphate-anchored biomolecule. The results demonstrated that the obtained nanocomposites could potentially be applied in transdermal drug delivery for anesthetics, analgesics and antibiotics.


Osseous spinal sarcoidosis: an unusual but important entity to remember.
Sarcoidosis is a multi-system disorder of unknown aetiology characterised by non-caseating granulomatous inflammation with varying presentation and prognosis. Bone involvement is uncommon and vertebral involvement is rare. We report a case of vertebral osseous sarcoidosis which presented with pulmonary symptoms mimicking tuberculosis and later developed vertebral involvement despite administration of oral corticosteroids.


Whey protein stimulates postprandial muscle protein accretion more effectively than do casein and casein hydrolysate in older men.
Sarcopenia has been attributed to a diminished muscle protein synthetic response to food intake. Differences in digestion and absorption kinetics of dietary protein, its amino acid composition, or both have been suggested to modulate postprandial muscle protein accretion. The objective was to compare protein digestion and absorption kinetics and subsequent postprandial muscle protein accretion after ingestion of whey, casein, and casein hydrolysate in healthy older adults. A total of 48 older men aged 74 ± 1 y (mean ± SEM) were randomly assigned to ingest a meal-like amount (20 g) of intrinsically l-[1-(13)C]phenylalanine-labeled whey, casein, or casein hydrolysate. Protein ingestion was combined with continuous intravenous l-[ring-(2)H(5)]phenylalanine infusion to assess in vivo digestion and absorption kinetics of dietary protein. Postprandial mixed muscle protein fractional synthetic rates (FSRs) were calculated from the ingested tracer. The peak appearance rate of dietary protein-derived phenylalanine in the circulation was greater with whey and casein hydrolysate than with casein (P < 0.05). FSR values were higher after whey (0.15 ± 0.02%/h) than after casein (0.08 ± 0.01%/h; P < 0.01) and casein hydrolysate (0.10 ± 0.01%/h; P < 0.05) ingestion. A strong positive correlation (r = 0.66, P < 0.01) was observed between peak plasma leucine concentrations and postprandial FSR values. Whey protein stimulates postprandial muscle protein accretion more effectively than do casein and casein hydrolysate in older men. This effect is attributed to a combination of whey's faster digestion and absorption kinetics and higher leucine content. This trial was registered at clinicaltrials.gov as NCT00557388.


Superstable Wet Foams and Lightweight Solid Composites from Nanocellulose and Hydrophobic Particles.
Colloids are suitable options to replace surfactants in the formation of multiphase systems while simultaneously achieving performance benefits. We introduce synergetic combination of colloids for the interfacial stabilization of complex fluids that can be converted into lightweight materials. The strong interactions between high aspect ratio and hydrophilic fibrillated cellulose (CNF) with low aspect ratio hydrophobic particles afford superstable Pickering foams. The foams were used as a scaffolding precursor of porous, solid materials. Compared to foams stabilized by the hydrophobic particles alone, the introduction of CNF significantly increased the foamability (by up to 350%) and foam lifetime. These effects are ascribed to the fibrillar network formed by CNF. The CNF solid fraction regulated the interparticle interactions in the wet foam, delaying or preventing drainage, coarsening, and bubble coalescence. Upon drying, such a complex fluid was transformed into lightweight and strong architectures, which displayed properties that depended on the surface energy of the CNF precursor. We show that CNF combined with hydrophobic particles universally forms superstable complex fluids that can be used as a processing route to synthesize strong composites and lightweight structures.


The hypoxia-inducible epigenetic regulators Jmjd1a and G9a provide a mechanistic link between angiogenesis and tumor growth.
Hypoxia promotes stem cell maintenance and tumor progression, but it remains unclear how it regulates long-term adaptation toward these processes. We reveal a striking downregulation of the hypoxia-inducible histone H3 lysine 9 (H3K9) demethylase JMJD1A as a hallmark of clinical human germ cell-derived tumors, such as seminomas, yolk sac tumors, and embryonal carcinomas. Jmjd1a was not essential for stem cell self-renewal but played a crucial role as a tumor suppressor in opposition to the hypoxia-regulated oncogenic H3K9 methyltransferase G9a. Importantly, loss of Jmjd1a resulted in increased tumor growth, whereas loss of G9a produced smaller tumors. Pharmacological inhibition of G9a also resulted in attenuation of tumor growth, offering a novel therapeutic strategy for germ cell-derived tumors. Finally, Jmjd1a and G9a drive mutually opposing expression of the antiangiogenic factor genes Robo4, Igfbp4, Notch4, and Tfpi accompanied by changes in H3K9 methylation status. Thus, we demonstrate a novel mechanistic link whereby hypoxia-regulated epigenetic changes are instrumental for the control of tumor growth through coordinated dysregulation of antiangiogenic gene expression.


Cancer-specific survival outcomes among patients treated during the cytokine era of kidney cancer (1989-2005): a benchmark for emerging targeted cancer therapies.
The management of renal cell carcinoma (RCC) is evolving toward less extirpative surgery and the use of targeted therapy. The authors set out to provide a benchmark against which emerging therapies should be measured. A prospective database including clinical and pathological variables for 1632 patients with RCC treated between 1989 and 2005 was queried. Patients were stratified using the University of California-Los Angeles Integrated Staging System (UISS) into low-, intermediate-, and high-risk groups. Disease-specific survival (DSS) was measured. Response to systemic therapy for patients with advanced disease was assessed. Nephrectomy was performed in 1492 patients. Overall 5-, 10-, and 15-year DSS was 55%, 40%, and 29%. For localized disease, 5- and 10-year DSS for UISS low-, intermediate-, and high-risk groups was 97% and 92%, 81% and 61%, and 62% and 41%, respectively. For metastatic disease, 5- and 10-year DSS for UISS low-, intermediate-, and high-risk groups was 41% and 31%, 18% and 7%, and 8% and 0%, respectively. Patients with metastatic disease receiving immunotherapy (n=453) had complete response in 7% (median survival [MS], 120+ months), partial response in 15% (MS, 42.8 months), stable disease in 33% (MS, 38.6 months), and progressive disease in 45% (MS, 11.6 months). Most patients with localized RCC do well with surgery alone, but effective adjuvant therapy is needed for patients identified as at high risk for recurrence. For advanced disease, newer targeted and potentially less toxic treatments should be at least as effective as those achieved with aggressive surgical resection and immunotherapy.


Comparative analysis of folate derived PET imaging agents with [(18)F]-2-fluoro-2-deoxy-d-glucose using a rodent inflammatory paw model.
Activated macrophages play a significant role in initiation and progression of inflammatory diseases and may serve as the basis for the development of targeted diagnostic methods for imaging sites of inflammation. Folate receptor beta (FR-β) is differentially expressed on activated macrophages associated with inflammatory disease states yet is absent in either quiescent or resting macrophages. Because folate binds with high affinity to FR-β, development of folate directed imaging agents has proceeded rapidly in the past decade. However, reports of PET based imaging agents for use in inflammatory conditions remain limited. To investigate whether FR-β expressing macrophages could be exploited for PET based inflammatory imaging, two separate folate-targeted PET imaging agents were developed, 4-[(18)F]-fluorophenylfolate and [(68)Ga]-DOTA-folate, and their ability to target activated macrophages were examined in a rodent inflammatory paw model. We further compared inflamed tissue uptake with 2-[(18)F]fluoro-2-deoxy-d-glucose ([(18)F]-FDG). microPET analysis demonstrated that both folate-targeted PET tracers had higher uptake in the inflamed paw compared to the control paw. When these radiotracers were compared to [(18)F]-FDG, both folate PET tracers had a higher signal-to-noise ratio (SNR) than [(18)F]-FDG, suggesting that folate tracers may be superior to [(18)F]-FDG in detecting diseases with an inflammatory component. Moreover, both folate-PET imaging agents also bind to FR-α which is overexpressed on multiple human cancers. Therefore, these folate derived PET tracers may also find use for localizing and staging FR(+) cancers, monitoring response to therapy, and for selecting patients for tandem folate-targeted therapies.


[Effect of adaptation to altitude hypoxia in early ontogeny on the indices of higher nervous activity].
Newborn male and female Wistar rats were adapted to hypoxia in a pressure chamber. Adaptation was started from the "altitude" of 1000 m, 1 hour daily, and then the time and the intensity of hypoxic actions were gradually increased: beginning from the 17th day the animals were subjected to adaptation to the "altitude" of 5000 m, for 5 hours, 5 days a week. The defence conditioned reflex of active avoidance was provoked in the animals after 2-month adaptation. A tendency to a more rapid elaboration of the reflex and a markedly enhanced degree of its retention in comparison with control was revealed in the adapted males. No significant changes in the elaboration and rentention of the reflex were found in the female animals adapted to hypoxia under analogous conditions.


Stereoselective release behaviors of imprinted bead matrices.
In this work, the stereoselective release behaviors of "low"-swelling molecularly imprinted polymer (MIP) bead matrices in pressed-coat tablet type were studied. Either R-propranolol selective MIP or S-propranolol selective MIP was combined with excipients and racemic propranolol and fabricated into the matrix. Subsequently, the release of different propranolol enantiomers from the matrices was examined. Also, the microscopic structure of the hydrated "low"-swelling MIP matrix was determined using a cryogenic scanning electron microscope in order to compare with that of the hydrated "high"-swelling MIP matrix. In vitro release profiles of the "low"-swelling matrices showed a difference in the release of enantiomers, in that the non-template isomer was released faster than the template isomer. However, in the last phase of dissolution this difference reduced and later reversed, resulting at last in the type of specificity being similar to that obtained previously with "high"-swelling MIP matrices. In summary, MIP beads can be fashioned into matrices and incorporated into different formulations to regulate the resultant stereoselectivity. From the behaviors of stereoselective release observed in MIP matrices, we can conclude that the enantioselective-controlled delivery mechanism of MIPs via formulations depends on the relative affinity of the enantiomer for the template sites, as well as the nature of the polymer, such as hydrophobicity and swellability.


Local structure controls the nonaffine shear and bulk moduli of disordered solids.
Paradigmatic model systems, which are used to study the mechanical response of matter, are random networks of point-atoms, random sphere packings, or simple crystal lattices; all of these models assume central-force interactions between particles/atoms. Each of these models differs in the spatial arrangement and the correlations among particles. In turn, this is reflected in the widely different behaviours of the shear (G) and compression (K) elastic moduli. The relation between the macroscopic elasticity as encoded in G, K and their ratio, and the microscopic lattice structure/order, is not understood. We provide a quantitative analytical connection between the local orientational order and the elasticity in model amorphous solids with different internal microstructure, focusing on the two opposite limits of packings (strong excluded-volume) and networks (no excluded-volume). The theory predicts that, in packings, the local orientational order due to excluded-volume causes less nonaffinity (less softness or larger stiffness) under compression than under shear. This leads to lower values of G/K, a well-documented phenomenon which was lacking a microscopic explanation. The theory also provides an excellent one-parameter description of the elasticity of compressed emulsions in comparison with experimental data over a broad range of packing fractions.


Calorie restriction increases insulin-stimulated tyrosine phosphorylation of insulin receptor and insulin receptor substrate-1 in rat skeletal muscle.
A moderate reduction in calorie intake (calorie restriction, CR) improves insulin-stimulated glucose transport in skeletal muscle. Therefore, we studied muscle insulin signalling in ad libitum (AL) and CR ( approximately 60% AL intake for 20 days) fed rats, which received a control injection (sterile water) or an insulin injection (30 U kg-1 body weight). In control (not insulin-treated) rats, there was no detectable tyrosine phosphorylation of insulin receptor (IR), regardless of diet; no diet effect on tyrosine phosphorylation of insulin receptor substrate-1 (IRS1) or IRS1-associated phosphatidylinositol 3-kinase (PI3K) protein and 21% higher IRS1-associated PI3K activity in AL vs. CR. In insulin-treated rats, tyrosine-phosphorylated IR was 79% higher for CR vs. AL; tyrosine-phosphorylated IRS1 was 109% higher for CR vs. AL; IRS1-associated PI3K protein and IRS1-associated PI3K activity were unaffected by diet. Calorie restriction amplifies early insulin signalling steps without changing IRS1-associated PI3K, suggesting enhanced glucose transport is mediated by altering: IRS1-PI3K localization, PI3K associated with proteins other than IRS1 or post-PI3K events.


Discovery of Amino Alcohols as Highly Potent, Selective, and Orally Efficacious Inhibitors of Leukotriene A4 Hydrolase.
The discovery of chiral amino alcohols derived from our previously disclosed clinical LTA4H inhibitor LYS006 is described. In a biochemical assay, their optical antipodes showed similar potencies, which could be rationalized by the cocrystal structures of these compounds bound to LTA4H. Despite comparable stabilities in liver microsomes, they showed distinct in vivo PK properties. Selective O-phosphorylation of the (R)-enantiomers in blood led to clearance values above the hepatic blood flow, whereas the (S)-enantiomers were unaffected and exhibited satisfactory metabolic stabilities in vivo. Introduction of two pyrazole rings led to compound (S)-2 with a more balanced distribution of polarity across the molecule, exhibiting high selectivity and excellent potency in vitro and in vivo. Furthermore, compound (S)-2 showed favorable profiles in 16-week IND-enabling toxicology studies in dogs and rats. Based on allometric scaling and potency in whole blood, compound (S)-2 has the potential for a low oral efficacious dose administered once daily.


Posttranscriptional Gene Regulation of T Follicular Helper Cells by RNA-Binding Proteins and microRNAs.
T follicular helper (Tfh) cells are critically involved in the establishment of potent antibody responses against infectious pathogens, such as viruses and bacteria, but their dysregulation may also result in aberrant antibody responses that frequently coincide with autoimmune diseases or allergies. The fate and identity of Tfh cells is tightly controlled by gene regulation on the transcriptional and posttranscriptional level. Here, we provide deeper insights into the posttranscriptional mechanisms that regulate Tfh cell differentiation, function, and plasticity through the actions of RNA-binding proteins (RBPs) and small endogenously expressed regulatory RNAs called microRNAs (miRNAs). The Roquin family of RBPs has been shown to dampen spontaneous activation and differentiation of naïve CD4+ T cells into Tfh cells, since CD4+ T cells with Roquin mutations accumulate as Tfh cells and provide inappropriate B cell help in the production of autoantibodies. Moreover, Regnase-1, an endoribonuclease that regulates a set of targets, which strongly overlaps with that of Roquin, is crucial for the prevention of autoantibody production. Interestingly, both Roquin and Regnase-1 proteins are cleaved and inactivated after TCR stimulation by the paracaspase MALT1. miRNAs are expressed in naïve CD4+ T cells and help preventing spontaneous differentiation into effector cells. While most miRNAs are downregulated upon T cell activation, several miRNAs have been shown to regulate the fate of these cells by either promoting (e.g., miR-17-92 and miR-155) or inhibiting (e.g., miR-146a) Tfh cell differentiation. Together, these different aspects highlight a complex and dynamic regulatory network of posttranscriptional gene regulation in Tfh cells that may also be active in other T helper cell populations, including Th1, Th2, Th17, and Treg.


Lincomycin and spectinomycin concentrations in liquid swine manure and their persistence during simulated manure storage.
Antimicrobials administered to livestock can be excreted up to 75% in the feces and urine. Liquid swine manure from confined animal feeding operations is generally retained in lagoon storage until it is applied as a nutrient source to crop and pasture land. Thus, the applied manure becomes a possible source of antimicrobials to aquatic ecosystems. In the prairie region of Canada, lincomycin and spectinomycin are two antimicrobials that are frequently administered to pigs for prevention of post-weaning diarrhea. In order to assess the potential for contamination of prairie wetlands, concentrations of both antimicrobials were monitored in liquid manure from a commercial-scale barn during a 5-week study, and their persistence during simulated manure storage was investigated. LC-MS/MS analysis of manure extracts showed that concentrations of lincomycin and spectinomycin in the accumulating liquid manure at the end of the study were equivalent to 32 and 3%, respectively, of the doses administered to weanling pigs in their feed. In a laboratory study in which lagoon storage was simulated at room temperature using fortified liquid manure, concentrations of both antimicrobials showed a rapid initial decrease during the first 6 days, followed by a much slower dissipation, over a period of 5 months. Such persistence indicates that lincomycin and spectinomycin may be present in lagoon manure when applied as an amendment to agricultural land since many lagoons are emptied every 6 months (early spring and late fall).


Osteon pullout in the equine third metacarpal bone: effects of ex vivo fatigue.
An important concept in bone mechanics is that osteons influence mechanical properties in several ways, including contributing to toughness and fatigue strength by debonding from the interstitial matrix so as to "bridge" developing cracks. Observations of "pulled out" osteons on fracture surfaces are thought to be indicative of such behavior. We tested the hypothesis that osteon pullout varies with mode of loading (fatigue vs. monotonic), cortical region, elastic modulus, and fatigue life. Mid-diaphseal beams from the dorsal, medial, and lateral regions of the equine third metacarpal bone were fractured in four point bending by monotonic loading to failure under deflection control, with or without 10(5) cycles of previous fatigue loading producing 5000 microstrain (15-20% of the expected failure strain) on the first cycle; or sinusoidal fatigue loading to failure, under load or deflection control, with the initial cycle producing 10,000 microstrain (30-40% of the expected failure strain). Using scanning electron microscopy, percent fracture surface area exhibiting osteon pullout (%OP.Ar) was measured. Monotonically loaded specimens and the compression side of fatigue fracture surfaces exhibited no osteon pullout. In load-controlled fatigue, pullout was present on the tension side of fracture surfaces, was regionally dependent (occurring to a greater amount dorsally), and was correlated negatively with elastic modulus and positively with fatigue life. Regional variation in %OP.Ar was also significant for the pooled (load and deflection controlled) fatigue specimens. %OP.Ar was nearly significantly greater in deflection controlled fatigue specimens than in load-controlled specimens (p=0.059). The data suggest that tensile fatigue loading of cortical bone eventually introduces damage that results in osteonal debonding and pullout, which is also associated with increased fatigue life via mechanisms that are not yet clear.


Is audiovisual integration subserved by the superior colliculus in humans?
The brain effectively integrates multisensory information to enhance perception. For example, audiovisual stimuli typically yield faster responses than isolated unimodal ones (redundant signal effect, RSE). Here, we show that the audiovisual RSE is likely subserved by a neural site of integration (neural coactivation), rather than by an independent-channels mechanism such as race models. This neural site is probably the superior colliculus (SC), because an RSE explainable by neural coactivation does not occur with purple or blue stimuli, which are invisible to the SC; such an RSE only occurs for spatially and temporally coincident audiovisual stimuli, in strict adherence with the multisensory responses in the SC of the cat. These data suggest that audiovisual integration in humans occurs very early during sensory processing, in the SC.


A comparison of pregnancy outcomes in Ghanaian women with varying dietary diversity: a prospective cohort study protocol.
Poor dietary intake during pregnancy can have negative repercussions on the mother and fetus. This study therefore aims to explore the dietary diversity (DD) of pregnant women and its associations with pregnancy outcomes among women in Northern Ghana. The main outcome variables to be measured are gestational weight gain and birth weight. A prospective cohort study design will be used and 600 pregnant women in their first trimester will be systematically recruited at health facilities and followed until delivery. In three follow-up visits after recruitment, information on sociodemographic and general characteristics, physical activity (International Physical Activity Questionnaire (IPAQ) short form, dietary intake (24-hour food recall), anthropometry and pregnancy outcomes will be collected. DD will be measured three times using the minimum DD-women (MDD-W) indicator and the mean of the three values overall will be used to determine low (<5 food groups) and high (≥5 food groups) DD. Data will be analysed using SPSS. Comparisons between groups (categorical data) will be made using the χ2 test for proportions, and t-tests and ANOVA will be performed on continuous variables. Regression analysis will be used to identify independent outcome predictors while controlling for possible confounding factors. The results may help to identify differences in DD between healthy and unhealthy pregnancy outcomes. The study protocol has been approved by the ethics committee of Tehran University of Medical Sciences and the ethical review committee of the Tamale Teaching Hospital. Written informed consent will be obtained from all subjects. The results will be published in due course.


Long-range focal collateralization of axons arising from corticocortical cells in monkey sensory-motor cortex.
Small extracellular injections of HRP were placed into a stratum of corticocortical axons situated immediately deep to area 3b of the monkey somatic sensory cortex. This stratum had previously been demonstrated to contain corticocortical fibers linking the cytoarchitectonic fields of the somatic sensory cortex to one another and certain of them to the motor cortex. This method resulted in extremely successful filling of pyramidal cells, their axons, collateral axon branches, and terminations in areas 3b, 1, and 2 posterior to the injection and in areas 3a and 4 anterior to it. The major finding was that cells with somata situated in any one of these fields and with principal axons traversing the injection site have long collaterals, primarily in layers III and V, which can extend throughout their own cytoarchitectonic field and into one or more other fields. In these fields they give off focused, columnlike concentrations of terminal boutons, which can be separated from one another by 800 micron or more. The anterogradely labeled, primary corticocortical fibers, traced forwards into areas 3a and 4, have virtually identical focal terminations. These findings indicate that interareal connectivity in the sensory-motor cortex can be effected by the axon branches of single cells rather than by separate groups of cells, and this may form a basis for the convergence of place and modality information on single cells in the sensorimotor cortex, a convergence that is not seen in the thalamic input to this cortex.


Linking time-varying symptomatology and intensity of infectiousness to patterns of norovirus transmission.
Norovirus (NoV) transmission may be impacted by changes in symptom intensity. Sudden onset of vomiting, which may cause an initial period of hyper-infectiousness, often marks the beginning of symptoms. This is often followed by: a 1-3 day period of milder symptoms, environmental contamination following vomiting, and post-symptomatic shedding that may result in transmission at progressively lower rates. Existing models have not included time-varying infectiousness, though representing these features could add utility to models of NoV transmission. We address this by comparing the fit of three models (Models 1-3) of NoV infection to household transmission data from a 2009 point-source outbreak of GII.12 norovirus in North Carolina. Model 1 is an SEIR compartmental model, modified to allow Gamma-distributed sojourn times in the latent and infectious classes, where symptomatic cases are uniformly infectious over time. Model 2 assumes infectiousness decays exponentially as a function of time since onset, while Model 3 is discontinuous, with a spike concentrating 50% of transmissibility at onset. We use Bayesian data augmentation techniques to estimate transmission parameters for each model, and compare their goodness of fit using qualitative and quantitative model comparison. We also assess the robustness of our findings to asymptomatic infections. We find that Model 3 (initial spike in shedding) best explains the household transmission data, using both quantitative and qualitative model comparisons. We also show that these results are robust to the presence of asymptomatic infections. Explicitly representing explosive NoV infectiousness at onset should be considered when developing models and interventions to interrupt and prevent outbreaks of norovirus in the community. The methods presented here are generally applicable to the transmission of pathogens that exhibit large variation in transmissibility over an infection.


[The course and surgical treatment of gastroduodenal ulcer in patients with diabetes mellitus].
The work deals with the features of the course of gastric and duodenal ulcers in diabetes mellitus and the frequency and pattern of the complications. The authors recommend criteria for preparation for surgical treatment, methods for compensation of the disturbances of carbohydrate metabolism, and the methods of postoperative management of such patients. The late-term results and analysis of treatment are discussed briefly. The coexistence of peptic ulcer and diabetes mellitus is a rare occurrence. During a 20-year period 31 patients were treated at the clinic, 21 of them underwent operation, in 5 patients an operation was considered inexpedient, another 5 patients refused to be operated on. The postoperative period was free of complications in 19 patients, there were no fatal outcomes.


Central coherence in adolescents with bulimia nervosa spectrum eating disorders.
Weak central coherence-a tendency to process details at the expense of the gestalt-has been observed among adults with bulimia nervosa (BN) and is a potential candidate endophenotype for eating disorders (EDs). However, as BN behaviors typically onset during adolescence it is important to assess central coherence in this younger age group to determine whether the findings in adults are likely a result of BN or present earlier in the evolution of the disorder. This study examines whether the detail-oriented and fragmented cognitive inefficiency observed among adults with BN is observable among adolescents with shorter illness duration, relative to healthy controls. The Rey-Osterrieth Complex Figure Test (RCFT) was administered to a total of 47 adolescents with DSM5 BN, 42 with purging disorder (PD), and 25 healthy controls (HC). Performance on this measure was compared across the three groups. Those with BN and PD demonstrated significantly worse accuracy scores compared to controls in the copy and delayed recall condition with a moderate effect size. These findings were exacerbated when symptoms of BN increased. Poorer accuracy scores reflect a fragmented and piecemeal strategy that interferes with visual-spatial integration in BN spectrum disorders. This cognitive inefficiency likely contributes to broad difficulties in executive functioning in this population especially in the context of worsening bulimic symptoms. The findings of this study support the hypothesis that poor global integration may constitute a cognitive endophenotype for BN.


Patient throughput times for orthopedic outpatients in a department of radiology: results of an interdisciplinary quality management program.
The purpose of this project was to employ quality management methods in order to decrease throughput times for orthopedic outpatients sent to the department of radiology. The following intervals were measured at the onset of the study and after 6 and 12 months: (a) between arrivals at outpatient clinic and radiology counter; (b) between arrival at radiology counter and time of last radiograph; and (c) between time of last radiograph and radiology report printing time. After the initial measurement, numerous changes were initiated both in radiology and in orthopedic surgery. The mean interval between arrival at the outpatient clinic and in radiology decreased by one third from 60 min during the first measurement to 40 (p < 0.001) and 41 min during the second and third measurement. The proportion of patients with total radiology times of more than 30 min decreased from 41 to 29 % between the first and third measurements (p < 0.001). The corresponding results for radiology times of more than 45 min were 17 and 11 % (p = 0.03). A standard type of quality management program can be employed successfully in order to reduce radiology throughput times for orthopedic outpatients.


The GH/IGF-1 axis in chronic heart failure.
The classic model of Chronic Heart Failure (CHF) is rooted in the overexpression of neurohormonal molecules. To complement this paradigm, increasing evidence indicates that a variety of hormones may be down-regulated in CHF patients. The list includes growth hormone (GH) and its tissue effector insulin-like growth factor-1 (IGF-1). The GH/IGF-1 axis regulates cardiac growth, stimulates myocardial contractility, and influences the vascular system. The relationship between the GH/IGF-1 axis and the cardiovascular system has been extensively demonstrated in numerous studies in animals models and confirmed by the cardiac derangements secondary to both GH excess and deficiency in humans. Impaired activity of the GH/IGF-1 axis in CHF has been described by several independent groups and includes a wide array of abnormalities, including low IGF-1 levels, GH deficiency (GHD), and GH resistance that may be related to the severity of heart disease. According to several observations, these derangements are associated with poor clinical status and outcome. Since the first study of GH therapy in CHF in 1996, several placebo-controlled trials have been conducted with conflicting results. These discordant findings are likely explained by the degree of CHF-associated GH/IGF-1 impairment that may impact on individual responsiveness to GH administration. Biological actions of GH and IGF-1, cardiovascular implication of GH deficiency and GH excess, relation between somatotrophic axis and CHF are discussed. Results from trials of GH therapy, emerging therapeutic strategies, safety issues, and lack in evidence are also reported.


Are animal models relevant to key aspects of human parturition?
Preterm birth remains the most serious complication of pregnancy and is associated with increased rates of infant death or permanent neurodevelopmental disability. Our understanding of the regulation of parturition remains inadequate. The scientific literature, largely derived from rodent animal models, suggests two major mechanisms regulating the timing of parturition: the withdrawal of the steroid hormone progesterone and a proinflammatory response by the immune system. However, available evidence strongly suggests that parturition in the human has significantly different regulators and mediators from those in most of the animal models. Our objectives are to critically review the data and concepts that have arisen from use of animal models for parturition and to rationalize the use of a new model. Many animal models have contributed to advances in our understanding of the regulation of parturition. However, we suggest that those animals dependent on progesterone withdrawal to initiate parturition clearly have a limitation to their translation to the human. In such models, a linear sequence of events (e.g., luteolysis, progesterone withdrawal, uterine activation, parturition) gives rise to the concept of a "trigger" mechanism. Conversely, we propose that human parturition may arise from the concomitant maturation of several systems in parallel. We have termed this novel concept "modular accumulation of physiological systems" (MAPS). We also emphasize the urgency to determine the precise role of the immune system in the process of parturition in situations other than intrauterine infection. Finally, we accentuate the need to develop a nonprimate animal model whose physiology is more relevant to human parturition. We suggest that the guinea pig displays several key physiological characteristics of gestation that more closely resemble human pregnancy than do currently favored animal models. We conclude that the application of novel concepts and new models are required to advance translational research in parturition.


Temporal lobe atrophy on MRI in Parkinson disease with dementia: a comparison with Alzheimer disease and dementia with Lewy bodies.
To investigate the extent of medial temporal lobe atrophy (MTA) on MRI in Parkinson disease (PD) with and without dementia compared with Alzheimer disease (AD) and dementia with Lewy bodies (DLB) and to determine whether MTA correlates with cognitive impairment in PD and PD dementia (PDD). Coronal T1-weighted MRI scans were acquired from control subjects (n = 39) and patients with PD (n = 33), PDD (n = 31), DLB (n = 25), and AD (n = 31), diagnosed according to standardized clinical diagnostic criteria. Cognitive function was assessed using the Cambridge Cognitive Examination (CAMCOG), and MTA was rated visually using a standardized (Scheltens) scale. More severe MTA was seen in PDD (p = 0.007), DLB (p < 0.001), and AD (p < 0.001) vs control subjects. PD subjects had greater hippocampal atrophy than control subjects (p = 0.015) but less than subjects with DLB and AD, though not with PDD. MTA correlated with CAMCOG score and memory scores in the DLB group and with age in control, PDD, and AD groups. There were no correlations between MTA and cognitive impairment in PD, PDD, and AD. PDD and DLB had a similar profile of cognitive impairment and MTA. Medial temporal lobe atrophy (MTA) was seen in cognitively intact older subjects with Parkinson disease (PD) and was not more pronounced in Parkinson disease dementia (PDD). Alzheimer disease (AD) and, to a lesser extent, dementia with Lewy bodies (DLB) showed more pronounced MTA. Results suggest early hippocampal involvement in PD and that when dementia develops in PD, anatomic structures apart from the hippocampus are predominantly implicated. Greater hippocampal involvement in AD vs PDD and DLB is consistent with clinical, cognitive, and pathologic differences between the disorders.


Antibacterial Honeycomb Scaffolds for Achieving Infection Prevention and Bone Regeneration.
Surgical site infection (SSI) is a severe complication associated with orthopedic bone reconstruction. For both infection prevention and bone regeneration, the framework surface of osteoconductive and bioresorbable scaffolds must be locally modified by minimum antibacterial substances, without sacrificing the osteoconductivity of the scaffold framework. In this study, we fabricated antibacterial honeycomb scaffolds by replacing carbonate apatite, which is the main component of the scaffold, with silver phosphate locally on the scaffold surface via dissolution-precipitation reactions. When the silver content was 9.9 × 10-4 wt %, the honeycomb scaffolds showed antibacterial activity without cytotoxicity and allowed cell proliferation, differentiation, and mineralization. Furthermore, the antibacterial honeycomb scaffolds perfectly prevented bacterial infection in vivo in the presence of methicillin-resistant Staphylococcus aureus, formed new bone at 2 weeks after surgery, and were gradually replaced with a new bone. Thus, the antibacterial honeycomb scaffolds achieved both infection prevention and bone regeneration. In contrast, severe infection symptoms, including abscess formation, osteolytic lesions, and inflammation, occurred 2 weeks after surgery when honeycomb scaffolds without silver phosphate modification were implanted. Nevertheless, the unmodified honeycomb scaffolds eliminated bacteria and necrotic bone through their scaffold channels, resulting in symptom improvement and bone formation. These results suggest that the honeycomb structure is inherently effective in hindering bacterial growth. This novel insight may contribute to the development of antibacterial scaffolds. Moreover, our modification method is useful for providing antibacterial activity to various biomaterials.


[Evaluation of activities and effectiveness of an asthmatic child program developed in primary care].
To evaluate activities and effectiveness of a programme of attention to asthmatic child, defined by a proper diagnostic process and therapeutic intervention, by the gain of educational goals and by reduction of number of asthma attacks. A before-after study Primary care The first 81 children attracted to the programme. Aged 5 to 13. Each child was periodically revised (mean time of monitoring 11 months and 24 days: interval 7-17 months). Intervention obtained significative results (p < 0.0001) in: lung function study and allergens detection, use of inhalatory way and peak flow meter. Asthma attacks were reduced (p < 0.0001): before 216, mean 2.66, SD 1.85, range 0-8; and after 117, mean 1.44, SD 1.27, range 0-5. 1. A systematic and organized intervention from Primary Care results effective to improve quality of life in asthmatic child (better diagnostic, more abilities, less asthma attacks). 2. It must be stated new strategies for reduction of smoking at home of these children.


Caffeine intake from coffee and tea and invasive breast cancer incidence among postmenopausal women in the Women's Health Initiative.
Research findings remain inconsistent whether caffeine consumption is associated with invasive breast cancer. We aimed to examine the association between caffeine intake from coffee and tea and incident invasive breast cancer among postmenopausal women. We included 79 871 participants in the Women's Health Initiative Observational Study in the current analysis. Incident invasive breast cancers were identified through September 30, 2015. Caffeine intake (mg/day) from caffeinated and decaffeinated coffee and tea was estimated based on self-reported frequency (cups/day) and average caffeine amount in each beverage. Cox proportional hazards regression models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). Subgroup analyses were conducted to explore whether associations of caffeine intake from coffee and tea with invasive breast cancer were different by age, race and ethnicity, smoking status, body mass index, history of hormone therapy use, alcohol intake and subtypes of breast cancer. During a median follow-up of 16.0 years, 4719 incident invasive breast cancers were identified. No significant association was found between caffeine intake from coffee and tea and invasive breast cancer incidence after adjusting for demographic, lifestyle and reproductive factors: HRs (95% CIs) for increasing quartiles of caffeine intake compared to the lowest were 1.03 (0.94, 1.12), 1.04 (0.95, 1.13) and 1.03 (0.94, 1.13), respectively (P-for-trend = .54). No significant associations of coffee and tea intake (cups/day) with overall breast cancer risk were found. Our findings are consistent with others showing no clear association of caffeine consumption with invasive breast cancer among postmenopausal women.


Testing allostatic load factor structures among adolescents: A structural equation modeling approach.
Allostatic load (AL) represents cumulative biological "wear and tear" that results from chronic stress exposure over time, ultimately increasing risk for chronic disease. A consensus is lacking regarding the best operationalization of AL, particularly for younger, less studied populations. The purpose of this study was to test multiple hypothesized factor structures for AL to determine the best measurement approach for adolescents. We analyzed biologic data for 1900 adolescents aged 12-18 from four waves (2003-2010) of the National Health and Nutrition Examination Survey. AL indicator variables included cardiovascular (systolic BP, creatinine), metabolic (HDL, LDL, triglycerides, insulin, fasting glucose, HA1C, body mass index [BMI], waist circumference), and immune (albumin, CRP, WBC, EBV) biomarkers. Structural equation modeling was used to test the fit of five hypothesized AL factor structures. The data best supported a unidimensional factor structure, where the AL construct directly influenced each of the indicator variables. All but two of the indicators (HDL and albumin) had positive factor loadings, thus, as AL increases the values for those indicators also increase. The best indicators for AL were those measuring metabolic dysregulation, with BMI and waist circumference having the highest factor loadings (0.95 and 0.982, respectively). BMI and waist circumference may be some of the earliest clinical signs of elevated AL that manifest among adolescents. Future research should aim to include neuroendocrine biomarkers in their AL measures to have a more robust estimation of AL in younger populations.


Effects and Stress-Relieving Mechanisms of Dark Tea Polysaccharide in Human HaCaT Keratinocytes and SZ95 Sebocytes.
A new skincare application scenario for dark tea, a unique and post-fermented tea popular in the health food industry, was developed in this paper. The effects of dark tea polysaccharide (DTP) on stress-induced skin problems and its mechanism of action were investigated by modeling cortisone-induced stress injury in human HaCaT keratinocytes and SZ95 sebaceous gland cells. The results showed a reduced cortisol conversion induced by cortisone under the action of DTP with a concentration of 200 μg/mL, probably by inhibiting the expression of the HSD11B1 enzyme. DTP was also able to suppress the cortisone-induced elevation of lipid levels in SZ95 sebocytes at this concentration. In addition, the composition and structure of DTP were verified by ultrafiltration, ultraviolet-visible spectrophotometry (UV-VIS), high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and infrared spectroscopy. In brief, DTP has a unique and significant stress-relieving effect, which provides new ideas for the development of new ingredients for the skin care industry.


Anterior segment abnormalities and angle-closure glaucoma in a family with a mutation in the BEST1 gene and Best vitelliform macular dystrophy.
To present the clinical and electrophysiological findings in four members of a family with Best vitelliform macular dystrophy (BVMD) and angle-closure glaucoma (ACG). Four members of a family with BVMD were examined clinically, including visual acuity, slit-lamp examination, biomicroscopy, Goldmann applanation tonometry and gonioscopy. Measurements of the anterior chamber depth and axial length, visual field, optical coherence tomography, full-field electroretinography, multifocal electroretinography and electrooculography were performed. In addition molecular genetic analysis of the bestrophin-1 gene (BEST1), the microphthalmia-associated transcription factor gene (MITF) and the cone-rod homeobox gene (CRX) were performed. Four family members with the c.253T>C p.Y85H mutation in the BEST1 gene and BVMD in different stages also exhibited anterior segment abnormalities such as shallow anterior chambers (two cases), and reduced axial lengths in all cases. Microphthalmos (axial length ≤ 20mm) was found in the index patient and in her son. Hyperopia was found in all four examined patients. Closed angles/narrow angles were observed in patients with microphthalmos. The index patient developed ACG at the age of 12 years. Her son inherited microphthalmos, severe hyperopia, and narrow angles. He is at risk of developing ACG. No pathogenic mutation of the MITF or the CRX genes was detected in the index patient. BVMD could be associated with anterior segment abnormalities such as shallow anterior chambers, closed/narrow anterior chamber angles and ACG. Ophthalmologists should be aware of the association between ACG and BVMD. Examination of the anterior segment, gonioscopy and intraocular pressure control are recommended in patients with BVMD.


Characterization of BPV-like DNA in equine sarcoids.
The DNA from equine sarcoid samples from New York State and Switzerland was isolated and probed with bovine papillomavirus type 1 (BPV-1) to determine if BPV genomes were present. Twelve of 13 sarcoids from New York State and 17/20 sarcoids from Switzerland contained DNA that hybridized to the BPV-1 probe. Restriction enzyme analysis of the positive samples demonstrated restriction fragment profiles characteristic of BPV-1 in 22 sarcoids and restriction fragment profiles characteristic of bovine papillomavirus type 2 (BPV-2) in 7 sarcoids. In addition, three tissues histologically diagnosed as pyogranulomatous dermatitis, fibropapilloma, and fibrosarcoma contained BPV-like DNA. Tissues with BPV-1-like and BPV-2-like DNA contained an average of 285.7 (21 to 808) and 125.8 (2 to 762) BPV-like genomes per cell, respectively. Minor differences in the restriction fragment profiles of the BPV-like DNA and evidence for partial BPV-like genomes were found in some sarcoids. BPV-like DNA was not detected in lymphocyte DNA from sarcoid-affected horses. These results confirm previous observations and support the hypothesis that bovine papillomavirus, or a very similar virus, is linked to the cause of equine sarcoid.


Making anti-cytokine therapy more selective: Studies in mice.
Cytokines are involved in a wide range of functions shaping the normal immune response, yet inflammatory changes in the immune system due to dysregulated cytokine signaling may lead to the induction of autoimmunity. Cytokine inhibitors have revolutionized the treatment of many autoimmune diseases in recent years. Systemic cytokine ablation, however, is often associated with the development of adverse side effects and some patients simply do not respond to therapy. TNF, IL-1 and IL-6 are the best characterized proinflammatory cytokines considered as the main therapeutic targets for the treatment of several autoimmune and inflammatory diseases. But can anti-cytokine therapy become more selective and thus more efficient? This mini-review discusses several recently emerging paradigms and summarizes current experimental attempts to validate them in mouse studies.


Respiratory response to tobacco dust exposure.
Results of a study of the respiratory responses of 318 nonsmoking female workers to long-term tobacco dust exposure are reported. The mean total tobacco dust concentrations ranged from 0.9 to 27.5 mg per m3; the respirable fraction, from 0.3 to 3.6 mg per m3. The mean length of exposure to tobacco dust was 14.9 years; 24 per cent of the workers had been exposed to tobacco dust for 20 years or more. Comparatively low prevalences of chronic respiratory symptoms were found, and only the prevalences of chest tightness and wheezing were significantly higher among workers exposed to tobacco dust than those of the control group (P less than 0.01). Calculating the expected 1-sec forced expiratory volume and forced vital capacity values by means of multiple linear regression equations, developed on the basis of data obtained in the 210 control subjects, revealed no significant differences between the measured and expected ventilatory capacity values among any of the groups of the workers observed. In contrast to negative findings with regard to chronic respiratory effects, significant acute decreases in ventilatory capacity during the work shift were recorded. No dose-response relationship was found between the level of exposure and the acute decreases in ventilatory capacity.


Anatomy of the distal radioulnar joint.
The distal radioulnar joint (DRUJ) is of the utmost importance in treatment of fractures of the distal radius. To improve the treatment of these fractures, knowledge of the anatomy and biomechanics of the DRUJ is essential. Three major findings from anatomic specimen studies of the DRUJ are discussed. The radius of the curvature of the sigmoid notch is 4-7 mm larger than that of the ulnar head, and consequently pronation and supination are a combined rotation and sliding movement in the DRUJ. The ulnar head has an average inclination of 20 degrees relative to the long axis of the ulna. Being parallel to the ulnar head, the sigmoid notch of the radius should have the same inclination. In the DRUJ, stability supported by joint surface architecture is minimal, and the ligament is therefore of particular importance to the stability.


Assay of 6-gingerol in CO2 supercritical fluid extracts of ginger and evaluation of its sustained release from a transdermal delivery system across rat skin.
Ginger has been widely used as healthy food condiment as well as traditional Chinese medicine since antiquity. Multiple potentials of ginger for treatment of various ailments have been revealed. However, the biological half-life of 6-gingerol (a principal pungent ingredient of ginger) is only 7.23 minutes while taken orally. Delivery of ginger compositions by routes other than oral have scarcely been reported. Therefore, we studied a noninvasive transdermal drug delivery system (TDDS) of ginger to bypass hepatic first pass metabolism, avoid gastrointestinal degradation and achieve long persistent release of effective compositions. After establishment of a HPLC analysis method of 6-gingerol, assays of 6-gingerol were performed to compare two kinds of ginger extracts. Then, the characteristics of transdermal delivery of 6-gingerol in TDDS were exhibited. The results showed that the contents of 6-gingerol in two kinds of ginger extracts were significantly different. The maximal delivery percentage of 6-gingerol across rat skin at 20 h was more than 40% in different TDDS formulations. TDDS may provide long-lasting delivery of ginger compounds.


The echinocandin antifungals: an overview of the pharmacology, spectrum and clinical efficacy.
For over four decades, the principal target of antifungal therapy has been the fungal cell membrane sterol ergosterol. Although this has proven to be a successful and relatively selective antifungal target, collateral toxicity to mammalian cells (amphotericin B) and drug interactions (azoles) have been by-products of agents that target the fungal cell membrane. In the 1970s, the echinocandins were identified during the screening of fungal fermentation products for new antibiotic agents. These agents were subsequently shown to inhibit production of beta(1,3)-glucan, a key structural component of the fungal cell wall. Subsequent chemical modification of these natural products has led to the development of safer, semi-synthetic beta(1,3)-glucan synthase inhibitors with enhanced microbiological and clinical efficacy against infections caused by Candida and Aspergillus species. In this review, the pharmacology, spectrum and clinical efficacy of the three leading beta(1,3)glucan synthase inhibitors (caspofungin, micafungin and anidulafungin), which have completed phase III clinical trials, will be discussed and a perspective for the role of these agents in the management of life-threatening mycoses will be offered.


A pH responsive complexation-based drug delivery system for oxaliplatin.
A responsive drug delivery system (DDS) for oxaliplatin (OX) has been designed with a view to overcoming several drawbacks associated with this anticancer agent, including fast degradation/deactivation in the blood stream, lack of tumor selectivity, and low bioavailability. The present approach is based on the direct host-guest encapsulation of OX by a pH-responsive receptor, carboxylatopillar[6]arene (CP6A). The binding affinities of CP6A for OX were found to be pH-sensitive at biologically relevant pH. For example, the association constant (Ka) at pH 7.4 [Ka = (1.02 ± 0.05) × 104 M-1] is 24 times larger than that at pH 5.4 [Ka = (4.21 ± 0.06) × 102 M-1]. Encapsulation of OX within the CP6A cavity did not affect its in vitro cytotoxicity as inferred from comparison studies carried out in several cancer cells (e.g., the HepG-2, MCF-7, and A549 cell lines). On the other hand, complexation by CP6A serves to increase the inherent stability of OX in plasma by 2.8-fold over a 24 h incubation period. The formation of a CP6A⊃OX host-guest complex served to enhance in a statistically significant way the ability of OX to inhibit the regrowth of sarcoma 180 (S180) tumors in Kunming (KM) mice xenografts. The improved anticancer activity observed in vivo for CP6A⊃OX is attributed to the combined effects of enhanced stability of the host-guest complex and the pH-responsive release of OX. Specifically, it is proposed that OX is protected as the result of complex formation and then released effectively in the acidic tumor environment.


Rational Design of Hybrid SARS-CoV-2 Main Protease Inhibitors Guided by the Superimposed Cocrystal Structures with the Peptidomimetic Inhibitors GC-376, Telaprevir, and Boceprevir.
SARS-CoV-2 main protease (Mpro) is a cysteine protease that mediates the cleavage of viral polyproteins and is a validated antiviral drug target. Mpro is highly conserved among all seven human coronaviruses, with certain Mpro inhibitors having broad-spectrum antiviral activity. In this study, we designed two hybrid inhibitors UAWJ9-36-1 and UAWJ9-36-3 based on the superimposed X-ray crystal structures of SARS-CoV-2 Mpro with GC-376, telaprevir, and boceprevir. Both UAWJ9-36-1 and UAWJ9-36-3 showed potent binding and enzymatic inhibition against the Mpro's from SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV-OC43, HCoV-NL63, HCoV-229E, and HCoV-HKU1. Cell-based Flip-GFP Mpro assay results show that UAWJ9-36-1 and UAWJ9-36-3 inhibited the intracellular protease activity of SARS-CoV-2 Mpro. In addition, UAWJ9-36-1 and UAWJ9-36-3 had potent antiviral activity against SARS-CoV-2, HCoV-OC43, HCoV-NL63, and HCoV-229E, with UAWJ9-36-3 being more potent than GC-376 in inhibiting SARS-CoV-2. Selectivity profiling revealed that UAWJ9-36-1 and UAWJ9-36-3 had an improved selectivity index over that of GC-376 against host cysteine proteases calpain I and cathepsin L, but not cathepsin K. The X-ray crystal structures of SARS-CoV-2 Mpro with UAWJ9-36-1 and UAWJ9-36-3 were both solved at 1.9 Å, which validated our design hypothesis. Overall, hybrid inhibitors UAWJ9-36-1 and UAWJ9-36-3 are promising candidates to be further developed as broad-spectrum coronavirus antivirals.


Engineered Tissue Models to Replicate Dynamic Interactions within the Hematopoietic Stem Cell Niche.
Hematopoietic stem cells are the progenitors of the blood and immune system and represent the most widely used regenerative therapy. However, their rarity and limited donor base necessitate the design of ex vivo systems that support HSC expansion without the loss of long-term stem cell activity. This review describes recent advances in biomaterials systems to replicate features of the hematopoietic niche. Inspired by the native bone marrow, these instructive biomaterials provide stimuli and cues from cocultured niche-associated cells to support HSC encapsulation and expansion. Engineered systems increasingly enable study of the dynamic nature of the matrix and biomolecular environment as well as the role of cell-cell signaling (e.g., autocrine feedback vs paracrine signaling between dissimilar cells). The inherent coupling of material properties, biotransport of cell-secreted factors, and cell-mediated remodeling motivate dynamic biomaterial systems as well as characterization and modeling tools capable of evaluating a temporally evolving tissue microenvironment. Recent advances in HSC identification and tracking, model-based experimental design, and single-cell culture platforms facilitate the study of the effect of constellations of matrix, cell, and soluble factor signals on HSC fate. While inspired by the HSC niche, these tools are amenable to the broader stem cell engineering community.


Detection of human immunodeficiency virus type 1 Nef and CD4 physical interaction in living human cells by using bioluminescence resonance energy transfer.
CD4 down-regulation by human immunodeficiency virus type 1 (HIV-1) Nef protein is a key function for virus virulence. This activity may be mediated by a direct Nef-CD4 interaction. We investigated the formation, in situ, of such a complex between proteins using bioluminescence resonance energy transfer technology and co-immunoprecipitations. Our data clearly demonstrate that Nef and CD4 interact in intact human cells. Moreover, our results clearly indicate that the dileucine motif of the CD4 cytoplasmic domain, critical for the Nef-induced CD4 down-regulation, is not implicated in the Nef/CD4 complex formation in the cellular context.


Identification and differential vulnerability of a neural network in sleep deprivation.
The study aimed to identify task-related brain activation networks whose change in expression exhibits subject differences as a function of differential susceptibility to sleep deprivation. Brain activity during a non-verbal recognition memory task was investigated in an event-related functional MRI paradigm both prior to and after 48 h of sleep deprivation. Nineteen healthy subjects participated. Regional covariance analysis was applied to data. An activation network pattern was identified whose expression decreased from pre- to post-sleep deprivation in 15 out 19 subjects (P < 0.05). Differential decrease in expression correlated with worsening performance in recognition accuracy (P < 0.05). Sites of de-activation were found in the posterior cerebellum, right fusiform gyrus and precuneus, and left lingual and inferior temporal gyri; increased activation was found in the bilateral insula, claustrum and right putamen. A network whose expression decreased after sleep deprivation and correlated with memory performance was identified. We conclude that this activation network plays a role in cognitive function during sleep deprivation.


Novel bone surrogates for cranial surgery training.
Parietal graft lifts are trained on human or animal specimens or are directly performed on patients without extensive training. In order to prevent harm to the patient resulting from fast rotating machinery tools, the surgeon needs to apply appropriate forces. Realistic haptics are essential to identify the varying parietal bone layers and to avoid a penetration of the brain. This however, requires experience and training. Therefore, in this study, bone surrogate materials were evaluated with the aim to provide an anatomically correct artificial skull cap with realistic haptic feedback for graft lift training procedures. Polyurethane composites made of calcium carbonate and calcium phosphate were developed and were used to create customized bone surrogates, imitating both cancellous and cortical bone. Mechanical properties of these surrogates were validated for drilling, milling and sawing by comparison with human parietal bones. For that, surgical tool tips were automatically inserted into artificial and human bones in a customized test bench and the maximum axial insertion forces were analyzed. Axial tool insertion measurements in human parietal bones resulted in mean maximum forces of 1.8±0.5N for drilling, 1.7±0.3N for milling and 0.9±0.1N for sawing. Calcium carbonate-based materials achieved higher forces than the human bone for drilling and milling, and lower forces for sawing. The calcium phosphate-based bone surrogates showed comparable axial insertions forces for all investigated tools and were identified as a suitable surrogate for drilling (p=0.87 and 0.41), milling (p=0.92 and 0.63) and sawing (p=0.11 and 0.76) of the cortical layer and the cancellous bone, respectively. In conclusion, our findings suggest, that a suitable material composition for artificial parietal bones has been identified, mimicking the properties of human bone during surgical machinery procedures. Thus, these materials are suitable for surgical training and education in simulator training.


Pediatric cartilaginous lateral femoral condyle Hoffa fracture: a case report and review of the literature.
Hoffa fractures are challenging coronally-oriented articular injuries of the femoral condyle. These fractures are rare in adults and extremely rare in the skeletally immature, with few cases reported in literature. To prevent mal- or non-union, Hoffa fractures require prompt surgical stabilisation with anatomic reduction and internal fixation. We discuss the case of a lateral distal femoral condyle cartilaginous Hoffa fracture in a ten-year-old male patient. The patient presented after a football non-contact "twist and pop" injury with radiographic imaging described as an osteochondritis dissecans lesion. An MRI was obtained which demonstrated a lateral distal femoral condyle osteochondral fracture. An operative plan was formulated to perform arthroscopic reduction and bio-compression screw fixation to minimize damage to the physis and surrounding tissues. Hyperflexion of the knee allowed for anatomic fracture reduction with the placement of 2 bio-compression screws serving as maintenance of fixation. The patient did well postoperatively and returned to full activity after 6 months. Hoffa fractures in the pediatric population are rare and can occur not only through bone but also through the thick chondral layer in younger patients. These are extremely difficult to diagnose through X-Ray alone. The prompt use of MRI imaging allows for operative fixation in a timely fashion, while an arthroscopic-only approach allows for minimal tissue damage. With an appropriate fracture type, hyper-flexion reduces and stabilizes the fracture, permitting the placement of minimally invasive bio-compression fixation.


Systemic hypoxia mimicry enhances axonal regeneration and functional recovery following peripheral nerve injury.
Despite the ability of peripheral nerves to regenerate after injury, failure occurs due to an inability of supporting cells to maintain growth, resulting in long-term consequences such as sensorimotor dysfunction and neuropathic pain. Here, we investigate the potential of engaging the cellular adaptive response to hypoxia, via inhibiting its negative regulators, to enhance the regenerative process. Under normoxic conditions, prolyl hydroxylase domain (PHD) proteins 1, 2, and 3 hydroxylate the key metabolic regulator hypoxia inducible factor 1α (HIF1α), marking it for subsequent proteasomal degradation. We inhibited PHD protein function systemically via either individual genetic deletion or pharmacological pan-PHD inhibition using dimethyloxalylglycine (DMOG). We show enhanced axonal regeneration after sciatic nerve crush injury in PHD1-/- mice, PHD3-/- mice, and in DMOG-treated mice, and in PHD1-/- and DMOG-treated mice a reduction in hypersensitivity to cooling after permanent sciatic ligation. Electromyographically, PHD1-/- and PHD3-/- mice showed an increased CMAP amplitude one-month post-injury, probably due to protection against denervation induced muscle atrophy, while DMOG-treated and PHD2+/- mice showed reduced latencies, indicating improved motor axon function. DMOG treatment did not affect the growth of dorsal root ganglion neurites in vitro, suggesting a lack of direct effects of DMOG on axonal regrowth. Enhanced regeneration in vivo was concurrent with an increase in macrophage density, and a shift in macrophage polarization state ratios (from M1-like toward M2-like) in DMOG-treated animals. These results indicate PHD proteins as a novel therapeutic target to improve regenerative and functional outcomes after peripheral nerve injury without manipulating molecular O2.


[Mitral valve prolapse: a benign abnormality?].
Mitral valve prolapse should be considered a benign condition as there are few complications. The natural history is not significantly different to that of a matched control population. There are potentially severe forms of the condition which are mainly observed in specialised centres and this introduces a bias in patient selection when compared with the general population. These forms are relatively rare in clinical practice and can be easily identified by simple echocardiographic criteria of valvular thickness. In any case, patients with mitral valve prolapse should be reassured in order not to create or unduly increase anxiety which in itself may cause incapacity.


DNA/long peptide vaccination against conserved regions of SIV induces partial protection against SIVmac251 challenge.
We recently developed a HIVconsv vaccine strategy, consisting of combined conserved regions of HIV-1, to adequately cover viral diversity. To evaluate efficacy in nonhuman primates, an equivalent SIV-derived immunogen SIVconsv was designed and delivered as plasmid DNA or synthetic long peptides. Rhesus macaques lacking protective MHC class I alleles Mamu-A*001 : 01, B*008 : 01, B*017 : 01 were immunized with either SIVconsv synthetic long peptides (S) alone or in combination with plasmid DNA encoding the same conserved regions (D) using SSS or DDSS regimens. The SIVconsv synthetic long peptide vaccine consisted of 46 approximately 30-amino acid-long peptides emulsified in Montanide ISA-720 and adjuvanted with pegylated type I interferon and imiquimod. Both SSS and DDSS regimens generated high frequencies of SIV-specific IFN-γ-producing cells comparable with reported adenoviral vector systems. Strong polyfunctional CD4⁺ T-cell and modest CD8⁺ T-cell responses were generated, which were of central memory T-cell phenotype. Furthermore, SIVconsv-specific antibody responses were induced capable of recognizing the Env glycoprotein. Eight weeks after the last immunization, control and SIVconsv-vaccinated animals were challenged intrarectally with 10 MID50 of pathogenic SIVmac251. Two out of six animals in the DDSS group were protected against infection, while all 14 animals in the SSS and two control groups were infected. Vaccine induced SIV-specific IgG responses in mucosal washes prechallenge were highest in the two protected animals. This study demonstrates that vaccine-elicited responses towards conserved regions can afford partial protection against a high-dose intrarectal SIVmac251 challenge.


Strength, character, and directionality of halogen bonds involving cationic halogen bond donors.
Halogen bonds involving cationic halogen bond donors and anionic halogen bond acceptors have recently been recognized as being important in stabilizing the crystal structures of many salts. Theoretical characterization of these types of interactions, most importantly in terms of their directionality, has been limited. Here we generate high-quality symmetry adapted perturbation theory potential energy curves of a H3N-C[triple bond, length as m-dash]C-Br+Cl- model system in order to characterize halogen bonds involving charged species, in terms of contributions from electrostatics, exchange, induction, and dispersion, with special emphasis on analyzing contributions that are most responsible for the directionality of these interactions. It is found that, as in the case of neutral halogen bonds, exchange forces are important contributors to the directionality of charged halogen bonds, however, it is also found that induction effects, which contribute little to the stability and directionality of neutral halogen bonds, play a large role in the directionality of halogen bonds involving charged species. Potential energy curves based on the ωB97X-D/def2-TZVP/C-PCM method, which includes an implicit solvation model in order to mimic the effects of the crystal medium, are produced for both the H3N-C[triple bond, length as m-dash]C-Br+Cl- model system and for the 4-bromoaniliniumCl- dimer, which is based on the real 4-bromoanilinium chloride salt, whose crystal structure has been determined experimentally. It is found that, within a crystal-like medium, charged halogen bond are significantly weaker than in the gas phase, having optimum interaction energies up to approximately -20 kcal mol-1.


A position paper on Physical & Rehabilitation Medicine programmes in post-acute settings. Union of European Medical Specialists Section of Physical & Rehabilitation Medicine (in conjunction with the European Society of Physical & Rehabilitation Medicine).
Physical & Rehabilitation Medicine (PRM) programmes in post-acute settings cover interventions for the rehabilitation of people with a variety of disabling health conditions. The setting of the intervention is more important than the timing and these programmes can be carried out in a variety of facilities. This paper describes the role of PRM services and of PRM specialists in delivering rehabilitation programmes to people, who have initially been admitted to hospital. The emphasis is on improving patients' activities and addressing participation issues. PRM programmes in post-acute settings provide a range of treatments and have a major influence in the long-term on the pace and extent of return of function and recovery from ill-health. This paper will define the meaning of post-acute settings and will describe the patient's journey through the post-acute setting. In particular, it addresses the standards of care across Europe that patients should expect. This paper also examines the general principles of funding such programmes within the context of different health care systems across Europe. Coordinated care improves outcomes and economic profiles for both payers and providers of services. This paper describes the value of PRM interventions and PRM specialist-led teams in promoting better outcomes for people with disabilities with complex needs.


Hsp90 binds and regulates Gcn2, the ligand-inducible kinase of the alpha subunit of eukaryotic translation initiation factor 2 [corrected].
The protein kinase Gcn2 stimulates translation of the yeast transcription factor Gcn4 upon amino acid starvation. Using genetic and biochemical approaches, we show that Gcn2 is regulated by the molecular chaperone Hsp90 in budding yeast Saccharomyces cerevisiae. Specifically, we found that (i) several Hsp90 mutant strains exhibit constitutive expression of a GCN4-lacZ reporter plasmid; (ii) Gcn2 and Hsp90 form a complex in vitro as well as in vivo; (iii) the specific inhibitors of Hsp90, geldanamycin and macbecin I, enhance the association of Gcn2 with Hsp90 and inhibit its kinase activity in vitro; (iv) in vivo, macbecin I strongly reduces the levels of Gcn2; (v) in a strain expressing the temperature-sensitive Hsp90 mutant G170D, both the accumulation and activity of Gcn2 are abolished at the restrictive temperature; and (vi) the Hsp90 cochaperones Cdc37, Sti1, and Sba1 are required for the response to amino acid starvation. Taken together, these data identify Gcn2 as a novel target for Hsp90, which plays a crucial role for the maturation and regulation of Gcn2.


Cognitive status of children treated with central nervous system prophylactic chemotherapy for acute lymphocytic leukemia.
Treatment-related cognitive impairments have been reported for survivors of childhood leukemia following prophylactic central nervous system (CNS) treatment with craniospinal radiation. We examined the neurocognitive status of 46 children with acute lymphocytic leukemia (ALL) to assess the impact of a regimen consisting of systemic chemotherapy and prophylactic CNS chemotherapy. By comparing three groups of ALL children (i.e., patients whose diagnosis was recent, patients 1 year postdiagnosis currently receiving CNS prophylactic chemotherapy, and off-therapy patients who had been treated with chemotherapy for 3 years) and their healthy siblings on measures of sequential and simultaneous processing, we were able to examine the effects of CNS prophylactic and systemic chemotherapy at various points during treatment. Results indicate that the children who had received a 3-year course of chemotherapy (off-therapy patients) were more impaired on tasks involving right-hemisphere simultaneous processing than were sibling controls or ALL children whose diagnosis was recent and whose treatment had just begun. Age at diagnosis did not interact with the effects of chemotherapy. These findings support the need for continued evaluation of cognitive functioning in ALL, children receiving CNS prophylactic chemotherapy to identify potential harmful neurocognitive sequelae of treatment.


The effect of smoking on the male excess of bladder cancer: a meta-analysis and geographical analyses.
Smoking is considered the primary risk factor for bladder cancer. Although smoking prevalence and bladder cancer incidence vary around the world, bladder cancer is on average 4 times more common in males than in females. This article describes the observed male-female incidence ratio of bladder cancer for 21 world regions in 2002 and 11 geographical areas during the time period 1970-1997. A meta-analysis, including 34 studies, was performed to ascertain the increased risk for bladder cancer in males and females when smoking. The summary odds ratios (SORs) calculated in the meta-analysis were used to estimate the male-female incidence ratio of bladder cancer that would be expected for hypothetical smoking prevalence scenarios. These expected male-female incidence ratios were compared with the observed ratios to evaluate the role of smoking on the male excess of bladder cancer. The male-female incidence ratio of bladder cancer was higher than expected worldwide and over time, based on a smoking prevalence of 75% in males, 10% in females and an increased risk (SOR) of bladder cancer associated with smoking of 4.23 for males and 1.35 for females, respectively. This implied that, at least in the Western world, smoking can only partially explain the difference in bladder cancer incidence. Consequently, other factors are responsible for the difference in bladder cancer incidence.


Collision Cross Section Prediction with Molecular Fingerprint Using Machine Learning.
High-resolution mass spectrometry is a promising technique in non-target screening (NTS) to monitor contaminants of emerging concern in complex samples. Current chemical identification strategies in NTS experiments typically depend on spectral libraries, chemical databases, and in silico fragmentation tools. However, small molecule identification remains challenging due to the lack of orthogonal sources of information (e.g., unique fragments). Collision cross section (CCS) values measured by ion mobility spectrometry (IMS) offer an additional identification dimension to increase the confidence level. Thanks to the advances in analytical instrumentation, an increasing application of IMS hybrid with high-resolution mass spectrometry (HRMS) in NTS has been reported in the recent decades. Several CCS prediction tools have been developed. However, limited CCS prediction methods were based on a large scale of chemical classes and cross-platform CCS measurements. We successfully developed two prediction models using a random forest machine learning algorithm. One of the approaches was based on chemicals' super classes; the other model was direct CCS prediction using molecular fingerprint. Over 13,324 CCS values from six different laboratories and PubChem using a variety of ion-mobility separation techniques were used for training and testing the models. The test accuracy for all the prediction models was over 0.85, and the median of relative residual was around 2.2%. The models can be applied to different IMS platforms to eliminate false positives in small molecule identification.


Effects of oral moisturising gel containing egg yolk antibodies against Candida albicans in older people.
The aim of this study was to evaluate the inhibitory effects of oral moisturising gel containing egg yolk antibody against Candida albicans (anti-CA IgY) in older people. Therefore, we measured the number of Candia CFU present on oral swabs at baseline and after using the gel. A randomised, double-blind, placebo-controlled trial was conducted among volunteers living in a nursing home in Japan. The participants were divided into two groups. The group 1 participants received oral care using an experimental oral moisturising gel with anti-CA IgY, and those in group 2 received oral care using a placebo oral moisturising gel without anti-CA IgY. The oral care was performed by care workers three times a day for 4 weeks. The participants' tongues were sampled using a swab method at baseline and after 2 and 4 weeks of using the oral gel, and the number of C. albicans, Candida tropicalis and Candida krusei colonies was counted. The baseline oral condition of the participants in the two groups did not differ significantly. The experimental gel significantly reduced the number of C. albicans colonies from baseline to after 4 weeks of using the oral gel; however, no significant reductions were observed in the number of C. tropicalis or C. krusei colonies. The use of oral moisturising gel containing anti-CA IgY for 1 month significantly reduces the number of C. albicans CFU present on swabs in older people.


The relationship between apelin and cardiac parameters in patients on peritoneal dialysis: is there a new cardiac marker?
Many markers have been proposed for CVD risk assessment in dialysis population. Apelin is a peptide that has roles in cardiovascular functions and volume regulation namely vasodilation, decreased blood pressure (BP), positive inotropic effect and inhibition of antidiuretic hormone release. The aim of this study was to examine relationship of apelin levels with echocardiographic findings and laboratory parameters related with cardiovascular function and bone mineral metabolism among peritoneal dialysis (PD) patients. This is a cross-sectional study in which chronic PD patients aged between 18 and 80 without active cardiac, infectious or malignant diseases and hypervolemia have been included. Apelin-36 levels and echocardiographic findings were recorded as well as clinical and laboratory data. Of the 53 patients, the mean age and female/male ratio was 52.8 ± 15.3 years and 30/23, respectively. Mean apelin level was 1.45 ± 0.37 ng/ml. Gender, drugs (renin-angiotensin-aldosteron inhibitors, statins), presence of left ventricular hypertrophy, diabetes mellitus, hypertension, hyperlipidemia and significant residual renal function did not affect apelin-36 levels. Apelin-36 was correlated negatively with age and left atrium diameter; and positively with diastolic BP, ejection fraction (EF), total cholesterol, LDL-cholesterol, HDL-cholesterol, parathyroid hormone and alkaline phosphatase (ALP) levels. Diastolic BP, LDL-cholesterol, ALP and EF were found to be the independent determinants of apelin-36 levels with linear regression analysis. Apelinergic system has important roles in volume regulation, cardiovascular functions, lipid metabolism and bone mineral disorders in PD patients. Prospective studies with large population are required.


Evaluation of fibromyalgia syndrome in patients with rosacea.
This study aims to investigate the frequency of fibromyalgia syndrome (FMS) in rosacea patients and the relationship between disease disability score of FMS and quality of life score of rosacea. This cross-sectional controlled clinical trial was performed between December 2017 and December 2018. One hundred female rosacea patients (mean age 43.2±10.1; range, 21 to 65 years) and 100 age- and sex-matched control subjects (mean age 41.2±11.1; range, 22 to 68 years) with no history of skin disease and systemic diseases including diabetes, cardiovascular, renal and hepatic diseases were recruited. Dermatology Life Quality Index (DLQI) scores were calculated using a 10-item self-administered questionnaire. The diagnosis of FMS was established according to 2010 American College of Rheumatology diagnostic criteria. Fibromyalgia Impact Questionnaire (FIQ) was used to determine the clinical severity and functional disability, while Visual Analog Scale (VAS) was used to determine pain severity in the patients with FMS. The frequency of FMS in patient group was significantly higher than control group (p=0.019). The mean duration of FMS in patient group was significantly higher than control group (p=0.001). There was no significant difference in terms of the age of onset of FMS, FIQ and VAS scores between groups (p=0.53, p=0.54, p=0.07, respectively). DLQI scores were significantly correlated with FIQ scores in the patient group (r=0.43, p=0.008). The frequency of FMS in rosacea patients was significantly higher than control subjects without any skin disease and there was a correlation between disability score of FMS and quality of life score of rosacea. Investigating fibromyalgia symptoms in rosacea patients may be helpful for providing patient-based therapeutic approaches where neurologically based treatments may also be beneficial for rosacea.


Biocompatible small peptide super-hydrogelators bearing carbazole functionalities.
For the first time we have introduced carbazole capping groups onto two short peptide sequences, namely a diphenylalanine dipeptide and a glycine-diphenylalanine tripeptide, giving compounds 1 and 2, respectively. Both molecules form hydrogels at low concentrations, as low as 0.03% (w/v) for 1- well within the range of supergelators. Both gelators are composed of small ca. 2 nm thick molecular fibres, as elucidated by AFM and are non-cytotoxic towards HeLa cells at concentrations which are at or above their minimum gelation concentration.


Feline hypertension: clinical findings and response to antihypertensive treatment in 30 cases.
Systolic hypertension was diagnosed in 30 cats. At diagnosis, 16 of those were found to be in chronic renal failure only, while five were azotaemic and either receiving treatment for hyperthyroidism (four cases) or were untreated hyperthyroid cases (one case). Two cases were untreated hyperthyroid cases with no evidence of azotaemia and the remaining seven cases had no definitive diagnosis of the underlying cause of their hypertension. The successful treatment used for the majority of cases was amlodipine, which lowered systolic blood pressure from 202.5+/-16.8 to 153.2+/-21.6 mmHg (mean+/-SD; n=29) within the first 50 days. Each case was followed for at least three months, or to the end of its natural life, and each cat was re-examined every six to eight weeks. Systolic blood pressure was kept below a target value of 165 mmHg in 58 per cent of cases treated for three months or longer. At the time of writing, 19 of the cases had died or been euthanased with a median treatment time of 203 days, one case was lost to follow-up and 10 cases were still alive, nine of which had been treated for six months or more. Amlodipine can be used for long-term control of feline systemic hypertension.


Induction of the P2z/P2X7 nucleotide receptor and associated phospholipase D activity by lipopolysaccharide and IFN-gamma in the human THP-1 monocytic cell line.
The activation of phospholipase D (PLD) was used as a marker for P2z nucleotide receptor (P2zR) activity in the human THP-1 monocytic cell line. While untreated THP-1 cells displayed little PLD response to the P2zR agonist 3'-O-(4-benzoyl)benzoyl (Bz)ATP, treatment with either IFN-gamma or bacterial LPS induced a BzATP-mediated stimulation of PLD activity. Treatment of cells with combinations of IFN-gamma and LPS resulted in synergistic induction. P2z receptors mediated these effects because: 1) reduction of extracellular divalent cation concentration increased agonist potency; 2) only BzATP or ATP acted as agonist nucleotides; 3) oxidized ATP, an inhibitor of the P2z receptor, abolished the response. The P2zR-stimulated PLD was rapidly activated (t(1/2) = 1.5 min), completely inhibited by KN-62, a calcium-calmodulin kinase II inhibitor, and only partially repressed by bisindolylmaleimide, a protein kinase C inhibitor. The P2zR-mediated PLD activity was distinguished from phorbol ester-stimulated PLD activity because the latter was slowly activated (t(1/2) > 15 min), unaffected by oxidized ATP or KN-62, and completely inhibited by bisindolylmaleimide. IFN-gamma and LPS treatment also synergistically induced P2zR-dependent changes in membrane permeability and cytolysis, as indicated by BzATP-mediated Ca2+ influx, ethidium bromide uptake, and lactate dehydrogenase release. Finally, IFN-gamma and LPS synergistically up-regulated mRNA encoding the P2X7 receptor, a recently cloned ATP-gated channel that exhibits a P2zR phenotype.


Sonic Hedgehog Signaling Pathway Mediates Proliferation and Migration of Fibroblast-Like Synoviocytes in Rheumatoid Arthritis via MAPK/ERK Signaling Pathway.
Fibroblast-like synoviocytes (FLSs) are the major effector cells that lead to rheumatoid arthritis (RA) synovitis and joint destruction. Our previous studies showed that Sonic Hedgehog (SHH) signaling pathway is involved in aberrant activation of RA-FLSs and inhibition of SHH pathway decreases proliferation and migration of RA-FLSs. The objective of this study was to investigate if the SHH pathway mediates proliferation and migration of RA-FLSs via the mitogen-activated protein kinases/extracellular signal-regulated kinases (MAPK/ERK) signaling pathway. SHH signaling was studied by using SHH agonist (Purmorphamine) and antagonist (Cyclopamine) targeting the Smoothened (SMO) in FLSs. U0126-EtOH was used to inhibit the MAPK/ERK signaling pathway. The phosphorylation of ERK 1/2 (p-ERKl/2) was examined by western blot. Cell viability was detected using cell proliferation and cytotoxicity kit-8 (CCK8), and cell cycle distribution and proliferating cells were evaluated by the flow cytometry. Cell migration was examined by Transwell assay. Results showed that, compared with the control group, Purmorphamine increased the levels of p-ERK1/2 in concentration-and time-dependent manners (P < 0.01). Co-treated with Purmorphamine and U0126-EtOH or Cyclopamine both decreased the levels of p-ERK1/2 (P < 0.05). RA-FLSs treated with Purmorphamine resulted in alteration of cell cycle distribution, increasing of proliferating cells, cell viability, and migration cells compared to controls (P < 0.01). However, the above phenomenon can be abolished by U0126-EtOH (P < 0.05). The findings suggest that SHH signaling pathway mediates proliferation and migration of RA-FLSs via MAPK/ERK pathway and may contribute to progression of RA. Targeting SHH signaling may have a therapeutic potential in patients with RA.


Computational insights into the different catalytic activities of CYP3A4 and CYP3A5 toward schisantherin E.
The cytochromes CYP3A4 and CYP3A5 share 84% sequence identity, but they exhibit different catalytic activities toward some substrates. Schisantherin E (SE) was recently identified as a selective substrate of CYP3A5, which exhibited catalytic efficiency that was more than 23 times higher than CYP3A4. At present, however, the structural determinants responsible for the different catalytic activities of the two enzymes toward SE have not been fully understood. In this study, a combination of molecular docking, molecular dynamic simulations, and binding free energy calculation was performed on the CYP3A4/CYP3A5-SE systems to investigate the issue. The results demonstrate that Ser119 in CYP3A4 and Glu374 in CYP3A5 formed direct hydrogen bonding with SE, respectively. Additionally, one water molecule located between the B-C loop and the I helix mediated different hydrogen-bonding networks between CYP3A4/3A5 and SE. The residue differences (Phe/Leu108 and Leu/Phe210) triggered the distinct conformational changes of the Phe-cluster residues, especially Phe213 and Phe215, which formed stronger hydrophobic interactions with SE in CYP3A5. The calculated binding free energies were consistent with the experimental results.


[Surgical treatment of secondary hyperparathyroidism: a systematic review of the literature].
Secondary hyperparathyroidism (HPT) has a high prevalence in renal patients. Secondary HPT results from disturbances in mineral homeostasis, particularly calcium, which stimulates the parathyroid glands, increasing the secretion of parathyroid hormone (PTH). Prolonged stimulation can lead to autonomy in parathyroid function. Initial treatment is clinical, but parathyroidectomy (PTx) may be required. PTx can be subtotal or total followed or not followed by parathyroid tissue autograft. We compared the indications and results of these strategies as shown in the literature through a systematic literature review on surgical treatment of secondary HPT presented in MedLine and LILACS from January 2008 to March 2014. The search terms were: hyperparathyroidism; secondary hyperparathyroidism; parathyroidectomy and parathyroid glands, restricted to research only in humans, articles available in electronic media, published in Portuguese, Spanish, English or French. We selected 49 articles. Subtotal and total PTx followed by parathyroid tissue autograft were the most used techniques, without consensus on the most effective surgical procedure, although there was a preference for the latter. The choice depends on surgeon's experience. There was consensus on the need to identify all parathyroid glands and cryopreservation of parathyroid tissue whenever possible to graft if hypoparathyroidism arise. Imaging studies may be useful, especially in recurrences. Alternative treatments of secondary HPT, both interventional and conservative, require further study.


Mechanisms of cardiovascular changes in an experimental model of syndrome X and pharmacological intervention on the renin-angiotensin-system.
Various cardiovascular risk factors and disease states similar to those present in type 2 diabetic patients also seem to be present in non-diabetic individuals. This cluster of risk factors has been called syndrome X, also known as metabolic cardiovascular syndrome or insulin resistance syndrome. Vascular wall components changes, including endothelial dysfunction and vascular smooth muscle cell (VSMC) migration and proliferation, could be involved in the cardiovascular alterations associated with this state. Fructose fed rats (FFR) provide a model of dietary-induced insulin resistance, which has been used to assess the pathophysiological mechanisms of the metabolic and cardiovascular changes associated to the syndrome X. FFR have hyperinsulinemia, insulin resistance (altered glucose tolerance test) and hypertriglyceridemia; they also develop moderate hypertension and cardiac hypertrophy. This has been confirmed in male rats of different strains, such as Wistar and Sprague-Dawley, chronically fed with a 60% fructose-chow or 10% fructose in the drinking water. At different levels of the cardiovascular system, FFR exhibit changes in the nitric oxide generation system and in primary cultured proliferation of VSMC from conduit and resistance arteries. These abnormalities were normalized by long-term treatment with pharmacological agents acting on the renin-angiotensin system (RAS), such as angiotensin converting-enzyme inhibitors or angiotensin-AT(1) receptor antagonists, that also lowered blood pressure to control levels and reversed cardiac hypertrophy. Evidence suggests an important role for the RAS in the pathogenic mechanisms involved in this model of syndrome X. Furthermore, beneficial pharmacological intervention seems to be mediated by AT(2) receptors and kinins.


Analysis of avian leukosis virus DNA and RNA in bursal tumours: viral gene expression is not required for maintenance of the tumor state.
Each of twelve tumors induced by either Rous-associated virus-1 or -2 (RAV-1 or RAV-2) contained a predominant population of cells with ALV proviruses integrated at common sites, consistent with a clonal origin. Seven of nine RAV-2-induced bursal tumors contained single proviruses, and all seven solitary proviruses had suffered deletions. The detailed structures of four of these proviruses show that major deletions had occurred near or at the 5' ends, spanning sequences potentially important in the production of viral RNA. One provirus also lacked most of the information coding for the replicative functions of the virus. Restriction maps suggest that these four proviruses were inserted in similar regions of the host genome. We have studied virus-specific RNA in four bursal tumors and four cell lines derived from bursal tumors. No normal viral RNA species were detectable in three tumors containing single aberrant proviruses. However, transcripts of 2.2. kb which reacted only with a hybridization probe specific for the 5' end of viral RNA were observed in one of these three tumors. Analogous species, varying in length from 1.5 to 6.0 kb, were observed in a fourth bursal tumor with multiple proviruses and in all four cell lines. (This tumor and the cell lines also contained normal species of ALV mRNA and apparently normal proviral DNA). The structures of the aberrant proviruses and the absence of normal viral RNA in some tumors indicate that expression of viral genes is not required for maintenance of the tumor phenotype. In at least some cases, the mechanism of oncogenesis may involve stimulation of transcription of flanking cellular sequences by a viral promoter.


Entropy redistribution controls allostery in a metalloregulatory protein.
Allosteric communication between two ligand-binding sites in a protein is a central aspect of biological regulation that remains mechanistically unclear. Here we show that perturbations in equilibrium picosecond-nanosecond motions impact zinc (Zn)-induced allosteric inhibition of DNA binding by the Zn efflux repressor CzrA (chromosomal zinc-regulated repressor). DNA binding leads to an unanticipated increase in methyl side-chain flexibility and thus stabilizes the complex entropically; Zn binding redistributes these motions, inhibiting formation of the DNA complex by restricting coupled fast motions and concerted slower motions. Allosterically impaired CzrA mutants are characterized by distinct nonnative fast internal dynamics "fingerprints" upon Zn binding, and DNA binding is weakly regulated. We demonstrate the predictive power of the wild-type dynamics fingerprint to identify key residues in dynamics-driven allostery. We propose that driving forces arising from dynamics can be harnessed by nature to evolve new allosteric ligand specificities in a compact molecular scaffold.


[Dr Teodor Gjurgjević, lawyer, diplomat, polyglot, and the collaborator of Andrija Štampar].
Dr Theodor Gjurgjević was born in Stubička Kaniža on 1 December 1909 and died in Zagreb on 20 March 1976. He finished high school in Zagreb in 1927 and obtained a doctoral degree from the Zagreb University Faculty of Law in 1933. In 1956, he obtained his second doctorate in modern history from the University of Oxford. He dedicated a large part of his career to international affairs: before and during World War II in the Yugoslav Ministry of Foreign Affairs in Belgrade and after the war at the Zagreb School of Public Health as a collaborator of Andrija Štampar. At the School he held different positions: senior officer, lecturer in postgraduate courses, assistant, counselor, and from the 1959 School secretary. After the Second World War, he was delegated by the Yugoslav government to participate in various expert groups. Thanks to his very broad education, especially his knowledge of many foreign languages, he was able to help in the international presentation and recognition of the School of Public Health in the extraordinary post-war conditions. He is credited for the establishment and operation of the European Association of Schools of Public Health (ASPHER). He was buried in the arcades of the Zagreb cemetery Mirogoj, reserved for prominent national figures.