Unique aspects of herbal whole system research.
Whole systems of healthcare offer unique methodological and theoretical challenges for researchers. Herbalism has its own set of methodological and philosophical research issues that are beyond those presented for whole system research in general. It was our objective to explore various conceptual and methodological issues surrounding whole system herbal research. An International Society for Complementary Medicine Research workshop, Challenges in Herbal Whole System Research, was presented. Starting from a definition of herbalism, the most important challenges to herbal whole system research were elicited with inputs from both the workshop presenters and the audience. Five major challenges unique to herbal whole system research were identified: (1) defining herbalists and herbalism, (2) the role of the natural products industry in herbal research, (3) designing placebos and delivering active herbal treatments as given by herbalists, (4) researching the herb as a living entity, and (5) designing trials to investigate and develop multicomponent herbal therapies. Unique methods and theoretical frameworks are required to design studies of herbalism. Solutions to these methodological challenges need to be addressed to conduct research that examines herbal systems of medicine versus conducting trials on individual herbs given out of their original therapeutic context.


Kinesin-5-dependent poleward flux and spindle length control in Drosophila embryo mitosis.
We used antibody microinjection and genetic manipulations to dissect the various roles of the homotetrameric kinesin-5, KLP61F, in astral, centrosome-controlled Drosophila embryo spindles and to test the hypothesis that it slides apart interpolar (ip) microtubules (MT), thereby controlling poleward flux and spindle length. In wild-type and Ncd null mutant embryos, anti-KLP61F dissociated the motor from spindles, producing a spatial gradient in the KLP61F content of different spindles, which was visible in KLP61F-GFP transgenic embryos. The resulting mitotic defects, supported by gene dosage experiments and time-lapse microscopy of living klp61f mutants, reveal that, after NEB, KLP61F drives persistent MT bundling and the outward sliding of antiparallel MTs, thereby contributing to several processes that all appear insensitive to cortical disruption. KLP61F activity contributes to the poleward flux of both ipMTs and kinetochore MTs and to the length of the metaphase spindle. KLP61F activity maintains the prometaphase spindle by antagonizing Ncd and another unknown force-generator and drives anaphase B, although the rate of spindle elongation is relatively insensitive to the motor's concentration. Finally, KLP61F activity contributes to normal chromosome congression, kinetochore spacing, and anaphase A rates. Thus, a KLP61F-driven sliding filament mechanism contributes to multiple aspects of mitosis in this system.


The biofilm matrix.
The microorganisms in biofilms live in a self-produced matrix of hydrated extracellular polymeric substances (EPS) that form their immediate environment. EPS are mainly polysaccharides, proteins, nucleic acids and lipids; they provide the mechanical stability of biofilms, mediate their adhesion to surfaces and form a cohesive, three-dimensional polymer network that interconnects and transiently immobilizes biofilm cells. In addition, the biofilm matrix acts as an external digestive system by keeping extracellular enzymes close to the cells, enabling them to metabolize dissolved, colloidal and solid biopolymers. Here we describe the functions, properties and constituents of the EPS matrix that make biofilms the most successful forms of life on earth.


Early posttransplantation hemoglobin level corresponds with chronic renal dysfunction in heart transplant recipients.
The risk factors for moderate or severe chronic renal dysfunction (MSCRD) among heart transplant recipients may be distinct from those previously recognized owing to recently improved clinical care. We examined the clinical records of 88 adult patients who underwent first heart transplantations from 2000 to 2005 and survived 2 years. MSCRD was defined as a glomerular filtration rate (GFR) <60 mL/min/1.73 m(2) at 2 years after transplantation. Fifty patients were included in the MSCRD group and 38 in the non-MSCRD group. Loss of renal function was observed largely during the first 9 months after transplantation in the MSCRD group. The pretransplantation GFR was lower in the MSCRD group. Besides older age in the MSCRD group, there were no differences in baseline characteristics, immunosuppressive regimens, incidences of acute rejection episodes, cardiac allograft vasculopathy, or severe infections. The MSCRD group showed permanent lower posttransplantation hemoglobin levels. In multivariate logistic regression analysis, recipient age, pretransplantation GFR, postoperative intensive care unit stay and hemoglobin level at 9 month were unfavorable factors for posttransplantation MSCRD. In addition to recipient age and pretransplantation GFR as well established risk factors, our results suggest a prognostic value of a low early hemoglobin level for the development of chronic renal dysfunction after heart transplantation.


Origin of green photoluminescence in four-ring bent-core molecules with ESIPT, selective sensing of zinc ions by turn-on emission and their liquid crystal properties.
Fluorescent four-ring symmetrical/unsymmetrical molecules containing alkyl chains of a varied number of -CH2- groups with a bent-core have been synthesized to explore their liquid crystalline (LC) and photophysical properties. Some of these molecules depending upon their alkyl chain length were found to exhibit B1 and B7 liquid crystalline phases and are characterized by various analytical techniques such as FT-IR, 1H NMR, mass, POM, DSC, single crystal XRD, etc. Crystal structure determination reveals the hydrogen bonded enol form of these molecules with non-planar bent-molecular geometry. Intramolecular hydrogen bonding was found to play an important role in the stabilization of these molecules and in the origin of their green photoluminescence (GPL). The photo-physical experimental results through various control experiments clearly demonstrate that the origin of the large Stokes shifted GPL of these molecules can be attributed to the excited state intramolecular proton transfer (ESIPT) process. The formation of various types of anionic species and their stability were explored through steady-state and time-resolved fluorescence measurements. These compounds are found to be good turn-on PL probes in the selective detection of zinc ions at the micromolar level. Upon binding of zinc ions with the bent-core molecule, the structural changes have been investigated through NMR spectroscopy.


Three-phase vaporization theory for laser-activated microcapsules.
Precision control of vaporization, both in space and time, is critical for numerous applications, including medical imaging and therapy, catalysis and energy conversion, and it can be greatly improved through the use of micro- or nano-sized light absorbers. Ultimately, optimization of these applications also requires a fundamental understanding of the vaporization process. Upon laser irradiation, polymeric microcapsules containing a dye can vaporize, leading to the growth of a vapor bubble that emits a strong acoustic signature. Here, we compare laser-activated capsules containing either a volatile or a non-volatile oil core. We theoretically explore the vaporization of the capsules based on a three-phase thermodynamics model, that accounts for the partial vaporization of both the surrounding fluid and the oil core as well as for the interaction between heat transfer and microbubble growth. The model is compared to ultra-high-speed imaging experiments, where we record the cavitation events. Theory and experiments are in convincing agreement.


Properties of triple shape memory composites prepared via polymerization-induced phase separation.
Research in the field of shape memory polymers has recently witnessed the introduction of increasing complexity of material response, including such phenomena as triple/multishape behavior, temperature memory, and reversible actuation. Ordinarily, such complexity in physical behaviour is achieved through comparable complexity in material composition and synthesis. Seeking to achieve a triple shape behaviour with a simple route to materials synthesis, we introduce here a method that utilizes polymerization induced phase separation (PIPS) to yield the requisite combination of microstructure and composition. Thus, two blends incorporating epoxy and poly(ε-caprolactone) were developed using commercially available reactants, one featuring a semicrystalline epoxy and the other featuring an amorphous epoxy. We show that both blends exhibited distinct transition temperatures and three modulus-temperature plateaus needed for triple shape behaviour. Despite these similarities, their physical character at room temperature is vastly different: the semicrystalline epoxy material is elastomeric and the amorphous epoxy material is highly stiff. Characterization of the triple shape behaviour revealed an ability of both systems to fix two separate deformations independently, one by PCL crystallization and a second one by epoxy crystallization or vitrification, and recover both programmed shapes separately upon heating. Given the simplicity of fabrication, we envision application as multi-shape coatings, adhesives, and films.


Effects of interventions promoting monitoring of medication use and brief messaging on medication adherence for people with Type 2 diabetes: a systematic review of randomized trials.
To assess the impact of interventions promoting the monitoring of medication use and brief messaging to support medication adherence in patients with Type 2 diabetes mellitus, and to investigate the extent of theory use to guide intervention development. We systematically searched for controlled trials, published from 1990 onwards in Medline, Embase, CINAHL, PsycINFO and the Cochrane library, that evaluated interventions based on monitoring and brief messaging to support medication adherence in patients with Type 2 diabetes, to examine the effectiveness of such interventions. A total of 11 trials, comparing 15 interventions, were identified. Only a small minority presented a low risk of bias. Three interventions were based on delivering brief messages, six were based on monitoring medication adherence, and six used both strategies. Messaging interventions included the use of short message service text messages, web-based feedback, and messages delivered through monitoring devices. Monitoring interventions included remote self-reporting of medication and telephone calls with healthcare staff. Improvements in medication adherence were observed in six interventions, although effect sizes were generally moderate. Only two interventions improved both adherence and clinical outcomes. A meta-analysis of five trials (eight interventions) combining monitoring and messaging strategies showed that the pooled difference in medication adherence between intervention and control was moderate and not statistically significant [standardized mean difference = 0.22 (95% CI -0.05; 0.49)]. Only four trials were based on explicit theoretical frameworks. Although interventions based on messaging and monitoring have the potential to improve medication adherence in patients with Type 2 diabetes, evidence of their efficacy is limited and additional high-quality, theory-based research is needed.


Absorbed in the task: Personality measures predict engagement during task performance as tracked by error negativity and asymmetrical frontal activity.
We hypothesized that interactions between traits and context predict task engagement, as measured by the amplitude of the error-related negativity (ERN), performance, and relative frontal activity asymmetry (RFA). In Study 1, we found that drive for reward, absorption, and constraint independently predicted self-reported persistence. We hypothesized that, during a prolonged monotonous task, absorption would predict initial ERN amplitudes, constraint would delay declines in ERN amplitudes and deterioration of performance, and drive for reward would predict left RFA when a reward could be obtained. Study 2, employing EEG recordings, confirmed our predictions. The results showed that most traits that have in previous research been related to ERN amplitudes have a relationship with the motivational trait persistence in common. In addition, trait-context combinations that are likely associated with increased engagement predict larger ERN amplitudes and RFA. Together, these results support the hypothesis that engagement may be a common underlying factor predicting ERN amplitude.


Prime Time: long-term sexual health outcomes of a clinic-linked intervention.
Evidence about long-term effects of preventive health services for youth with complex needs is lacking. Prime Time, a youth development intervention, aims to reduce pregnancy risk among vulnerable adolescent females seeking clinic services. In a randomized trial, 253 sexually active females aged 13-17 who were at high risk for pregnancy were assigned to the Prime Time intervention or usual clinic services. The 18-month intervention, initiated in 2007-2008, comprised regular meetings with case managers and participation in youth leadership groups. Trial participants completed surveys at baseline and 30 months. Regression analyses were used to evaluate differences between groups in sexual and psychosocial outcomes at follow-up. At 30 months, the intervention group reported more months of consistent condom use (adjusted means, 1.8 vs. 1.1) and dual contraceptive use (0.9 vs. 0.3) in the past seven months than did controls. The intervention was most effective in promoting consistent use among participants with relatively high levels of connectedness to family or school. Fifteen percent of intervention participants, but only 6% of controls, reported having abstained from sex in the past six months (adjusted odds ratio, 2.9). Moreover, among high school graduates, those in the intervention group were more likely than those in the control group to have enrolled in college or technical school (72% vs. 37%; odds ratio, 4.5). Health services grounded in a youth development framework can lead to reductions in sexual risk among vulnerable youth that are evident one year following conclusion of services.


Molecular characterization of pyraclostrobin resistance and structural diversity of the cytochrome b gene in Botrytis cinerea from apple.
Botrytis cinerea isolates obtained from apple orchards were screened for resistance to the quinone outside inhibitor (QoI) pyraclostrobin. Of the 220 isolates tested, 43 (19.5%) were resistant to pyraclostrobin. Analysis of partial sequences of the cytochrome b gene (cyt b) in five pyraclostrobin-resistant (PR) and five pyraclostrobin-sensitive (PS) isolates showed that PR isolates harbored the point mutation leading to the substitution of glycine by alanine at codon position 143 in cyt b (G143A). Two pairs of allele-specific primers were designed based on this point mutation, and allele-specific polymerase chain reaction analysis with these primers showed that all 73 PR isolates (including 30 collected from decayed apple fruit) harbored the G143A mutation but PS isolates did not. Six pairs of primers were designed to analyze the presence of various introns in cyt b. There were six types (I to VI) of cyt b present in 247 isolates of B. cinerea collected from various apple-production areas in Washington State. Of the 247 isolates, 23 had type I cyt b containing all four introns (Bcbi-67/68, Bcbi-131/132, Bcbi-143/144, and Bcbi-164), 176 had type II cyt b containing three introns (Bcbi-67/68, Bcbi-131/132, and Bcbi-164), six had type III cyt b containing two introns (Bcbi-67/68 and Bcbi-131/132), one had type IV cyt b containing two introns (Bcbi-131/132 and Bcbi-164), one had type V cyt b containing only the Bcbi-131/132 intron, and 40 had type VI cyt b containing no introns. This is the first report of types III to VI cyt b present in B. cinerea. All 73 PR isolates did not carry the Bcbi-143/144 intron in cyt b. Of the 247 isolates tested, >90% did not carry the Bcbi-143/144 intron in cyt b, suggesting that B. cinerea populations from apple pose a high inherent risk for the development of resistance to QoIs because the presence of this intron in cyt b prevents the occurrence of G143A-mediated resistance. Analysis of genetic background based on three microsatellite primers showed that PR isolates originated from different lineages, and there was no correlation between cyt b types (I, II, and III) and the genetic background of the isolates; however, isolates carrying type VI cyt b might originate from the same lineage.


New serum-derived albumin scaffold seeded with adipose-derived stem cells and olfactory ensheathing cells used to treat spinal cord injured rats.
Recent advances in spinal cord injury (SCI) research and cell culture techniques and biomaterials predict promising new treatments for patients with SCI or other nerve injuries. Biomaterial scaffolds form a substrate within which cells are instructed to form a tissue in a controlled manner. This study was designed to assess axon regeneration and locomotor recovery in rats with spinal cord injury treated with a novel serum-derived albumin scaffold seeded with adipose derived stem cells (ADSCs) and olfactory ensheathing cells (OECs). OECs are considered promising candidates for the treatment of SCI, and ADSCs have the ability to differentiate into neural lineages. In vitro experiments revealed that ADSCs and OECs adhered to the scaffold, remained viable and expressed specific markers of their cell types when cultured in the scaffold. Rats treated with scaffold plus cells showed locomotor skills at several time points from 45 days post-injury that were improved over those recorded in control injured, untreated animals. Astrocytic scars and tissue regeneration, identified using histological and immunohistochemical techniques, revealed that although the scaffold itself appeared to play a significant role in reducing glial scar formation and filling of the lesion cavity with cells, the presence of ADSCs and OECs in the scaffold led to the appearance of cells expressing markers of neurons and axons at the injury site. Our findings point to the clinical feasibility of an albumin scaffold seeded with ADSCs and OECs as a treatment candidate for use in spinal cord injury repair studies.


Etiopathogenesis of Graves' disease.
Graves' disease is an autoimmune disorder, caused by thyroid-stimulating antibodies, which bind to and activate the thyrotropin receptor on thyroid cells, inducing the synthesis and release of thyroid hormones. It is a polygenic and multifactorial disease that develops as a result of complex interaction between genetic susceptibility and environmental and/or endogenous factors. Graves' disease differs from other autoimmune diseases of the thyroid by specific clinical features, including hyperthyroidism, vascular goitre, ophthalmopathy and--less commonly--infiltrative dermopathy. This article discusses current theories, regarding the etiology and pathogenesis of Graves' disease, including possible predisposing factors, autoimmune aspects of Graves' disease, ophthalmopathy, and dermopathy.


Ocular hypotensive effects of topically administered agmatine in a chronic ocular hypertensive rat model.
Agmatine, a primary polyamine and potential neuromodulator, exhibits a high affinity to the alpha2-adrenergic receptor as well as imidazoline receptors. As alpha2-adrenergic receptor agonists display positive ocular hypotensive effects, we assessed whether agmatine effectively lowers intraocular pressure (IOP) using a chronic ocular hypertensive rat model. We raised IOP in unilateral eyes of Sprague-Dawley rats by cauterizing three episcleral veins per eye. Four weeks later, we topically administered 10(-3) M agmatine solution 4 times a day for 6 consecutive weeks. After confirming the recovery of IOP to pretreatment level at 13 weeks after cauterization, the retinal ganglion cells (RGCs) were retrogradely labeled and counted. Eyes subjected to episcleral vein cauterization (EVC) demonstrated significant increases in IOP (48.39% increase over baseline IOP), and the elevated IOP was well maintained until 12 weeks. Topically administered agmatine powerfully lowered IOP to 30.29% of its pretreatment level, and the associated washout period was about two weeks. EVC was associated with a 55.44% loss of RGCs in the control group, but agmatine appeared to attenuate this RGC loss to 18.65%. Overall, topically administered agmatine appeared to effectively lower IOP and rescue RGCs in a chronic ocular hypertensive rat model. Although the mechanism underlying these effects is not yet established, it is possible that agmatine offers a powerful new ocular hypotensive agent for eyes with chronic ocular hypertension and/or glaucoma.


Lactobacillus plantarum PS128 Improves Physiological Adaptation and Performance in Triathletes through Gut Microbiota Modulation.
A triathlon is an extremely high-intensity exercise and a challenge for physiological adaptation. A triathlete's microbiome might be modulated by diet, age, medical treatments, lifestyle, and exercise, thereby maintaining aerobiosis and optimum health and performance. Probiotics, prebiotics, and synbiotics have been reported to have health-promoting activities (e.g., immunoregulation and cancer prevention). However, few studies have addressed how probiotics affect the microbiota of athletes and how this translates into functional activities. In our previous study, we found that Lactobacillus plantarum PS128 could ameliorate inflammation and oxidative stress, with improved exercise performance. Thus, here we investigate how the microbiota of triathletes are altered by L. plantarum PS128 supplementation, not only for exercise performance but also for possible physiological adaptation. The triathletes were assigned to two groups: an L. plantarum 128 supplement group (LG, 3 × 1010 colony-forming units (CFU)/day) and a placebo group (PG). Both groups continued with their regular exercise training for the next 4 weeks. The endurance performance, body composition, biochemistries, blood cells, microbiota, and associated metabolites were further investigated. PS128 significantly increased the athletes' endurance, by about 130% as compared to the PG group, but there was no significant difference in maximal oxygen consumption (VO2max) and composition between groups. The PS128 supplementation (LG) modulated the athlete's microbiota with both significant decreases (Anaerotruncus, Caproiciproducens, Coprobacillus, Desulfovibrio, Dielma, Family_XIII, Holdemania, and Oxalobacter) and increases (Akkermansia, Bifidobacterium, Butyricimonas, and Lactobacillus), and the LG showed lower diversity when compared to the PG. Also, the short-chain fatty acids (SCFAs; acetate, propionate, and butyrate) of the LG were significantly higher than the PG, which might be a result of a modulation of the associated microbiota. In conclusion, PS128 supplementation was associated with an improvement on endurance running performance through microbiota modulation and related metabolites, but not in maximal oxygen uptake.


ω-Alkynyl lipid surrogates for polyunsaturated fatty acids: free radical and enzymatic oxidations.
Lipid and lipid metabolite profiling are important parameters in understanding the pathogenesis of many diseases. Alkynylated polyunsaturated fatty acids are potentially useful probes for tracking the fate of fatty acid metabolites. The nonenzymatic and enzymatic oxidations of ω-alkynyl linoleic acid and ω-alkynyl arachidonic acid were compared to that of linoleic and arachidonic acid. There was no detectable difference in the primary products of nonenzymatic oxidation, which comprised cis,trans-hydroxy fatty acids. Similar hydroxy fatty acid products were formed when ω-alkynyl linoleic acid and ω-alkynyl arachidonic acid were reacted with lipoxygenase enzymes that introduce oxygen at different positions in the carbon chains. The rates of oxidation of ω-alkynylated fatty acids were reduced compared to those of the natural fatty acids. Cyclooxygenase-1 and -2 did not oxidize alkynyl linoleic but efficiently oxidized alkynyl arachidonic acid. The products were identified as alkynyl 11-hydroxy-eicosatetraenoic acid, alkynyl 11-hydroxy-8,9-epoxy-eicosatrienoic acid, and alkynyl prostaglandins. This deviation from the metabolic profile of arachidonic acid may limit the utility of alkynyl arachidonic acid in the tracking of cyclooxygenase-based lipid oxidation. The formation of alkynyl 11-hydroxy-8,9-epoxy-eicosatrienoic acid compared to alkynyl prostaglandins suggests that the ω-alkyne group causes a conformational change in the fatty acid bound to the enzyme, which reduces the efficiency of cyclization of dioxalanyl intermediates to endoperoxide intermediates. Overall, ω-alkynyl linoleic acid and ω-alkynyl arachidonic acid appear to be metabolically competent surrogates for tracking the fate of polyunsaturated fatty acids when looking at models involving autoxidation and oxidation by lipoxygenases.


Altered expression of Na(+)/K(+)-ATPase and other osmoregulatory genes in the gills of euryhaline animals in response to salinity transfer: a meta-analysis of 59 quantitative PCR studies over 10 years.
Recent advances in molecular techniques have allowed gene expression in euryhaline animals to be quantified during salinity transfers. As these investigations transition from studying single genes to utilizing genomics-based methodologies, it is an appropriate time to summarize single gene studies. Therefore, a meta-analysis was performed on 59 published studies that used quantitative polymerase chain reaction (qPCR) to examine expression of osmoregulatory genes (the Na(+)/K(+)-ATPase, NKA; the Na(+)/K(+)/2Cl(-) cotransporter, NKCC; carbonic anhydrase, CA; the cystic fibrosis transmembrane regulator, CFTR; and the H(+)-ATPase, HAT) in response to salinity transfer. Based on 887 calculated effect sizes, NKA, NKCC, CA, and HAT are up-regulated after salinity transfer, while surprisingly, CFTR is unchanged. Meta-analysis also identified influential factors contributing to these changes. For example, expression was highest: 1) during transfers from higher to lower salinities comprising a physiological transition from osmoconformity to osmoregulation, 2) 1-3 days following transfer, 3) during dissimilar transfers, and 4) in crustaceans rather than teleosts. Methodological characteristics (e.g., types of controls) were not important. Experiments lacking in the current literature were also identified. Meta-analyses are powerful tools for quantitatively synthesizing a large body of literature, and this report serves as a template for their application in other areas of comparative physiology.


Open-source sensor for measuring oxygen partial pressures below 100 microbars.
The ability to measure partial pressures of oxygen below 100 microbars and nanomolar dissolved oxygen concentrations in in situ laboratory systems benefits many fields including microbiology, geobiology, oceanography, chemistry, and materials science. Here, we present an easily constructible open-source design for a networked luminescence lifetime measurement system for in situ measurements in arbitrary laboratory containers. The system is well suited for measuring oxygen partial pressures in the 0-100 μbar range, with the maximum potentially usable upper range limit at around 10 mbar, depending on experimental conditions. The sensor has a limited drift and its detectability limit for oxygen is at 0.02 μbar for short timescale measurements. Each sensor can connect to a Wi-Fi network and send the logged data either over the Internet or to a local server, enabling a large number of parallel unattended experiments. Designs are also provided for attaching the sensor to various commercially available containers used in laboratories. The design files are released under an open source license, which enables other laboratories to build, customize, and use these sensors.


Mitotic control of human papillomavirus genome-containing cells is regulated by the function of the PDZ-binding motif of the E6 oncoprotein.
The function of a conserved PDS95/DLG1/ZO1 (PDZ) binding motif (E6 PBM) at the C-termini of E6 oncoproteins of high-risk human papillomavirus (HPV) types contributes to the development of HPV-associated malignancies. Here, using a primary human keratinocyte-based model of the high-risk HPV18 life cycle, we identify a novel link between the E6 PBM and mitotic stability. In cultures containing a mutant genome in which the E6 PBM was deleted there was an increase in the frequency of abnormal mitoses, including multinucleation, compared to cells harboring the wild type HPV18 genome. The loss of the E6 PBM was associated with a significant increase in the frequency of mitotic spindle defects associated with anaphase and telophase. Furthermore, cells carrying this mutant genome had increased chromosome segregation defects and they also exhibited greater levels of genomic instability, as shown by an elevated level of centromere-positive micronuclei. In wild type HPV18 genome-containing organotypic cultures, the majority of mitotic cells reside in the suprabasal layers, in keeping with the hyperplastic morphology of the structures. However, in mutant genome-containing structures a greater proportion of mitotic cells were retained in the basal layer, which were often of undefined polarity, thus correlating with their reduced thickness. We conclude that the ability of E6 to target cellular PDZ proteins plays a critical role in maintaining mitotic stability of HPV infected cells, ensuring stable episome persistence and vegetative amplification.


Isolation of mesenchymal stem cells from Pap smear samples.
Exploiting their ability to differentiate into mesenchymal lineages like cartilage, bone, fat, and muscle, and to elicit paracrine effects, mesenchymal stem cells (MSCs) are widely used in clinical settings to treat tissue injuries and autoimmune disorders. One of accessible sources of MSC is the samples used for Papanicolaou (Pap) test, which is a cervical screening method for detecting potentially pre-cancerous and cancerous alterations in the cervical cells and to diagnose genetic abnormalities in fetuses. This study aimed to identify and isolate the stem cells from Pap smear samples collected from pregnant women, and to trace the origin of these cells to maternal or fetal tissue, and characterize their stem cell properties. To investigate the possibility and efficiency of establishing MSC lines from the Pap smear samples, we were able to establish 6 cell lines from Pap smear samples from 60 pregnant women at different stages of gestation. The 3 cell lines randomly selected among the 6 established in this study, displayed high proliferation rates, several characteristics of MSCs, and the capacity to differentiate into adipocytes, osteocytes, and chondrocytes. Our study identified that the stem cell lines obtainable from Pap smear sampling were uterine cervical stromal cells (UCSCs) and had 10% efficiency of establishment. Despite their low efficiency of establishment, human UCSCs from Pap smear samples can become a simple, safe, low-cost, and donor-specific source of MSCs for stem cell therapy and regenerative medicine.


A lightweight tangible 3D interface for interactive visualization of thin fiber structures.
We present a prop-based, tangible interface for 3D interactive visualization of thin fiber structures. These data are commonly found in current bioimaging datasets, for example second-harmonic generation microscopy of collagen fibers in tissue. Our approach uses commodity visualization technologies such as a depth sensing camera and low-cost 3D display. Unlike most current uses of these emerging technologies in the games and graphics communities, we employ the depth sensing camera to create a fish-tank stereoscopic virtual reality system at the scientist's desk that supports tracking of small-scale gestures with objects already found in the work space. We apply the new interface to the problem of interactive exploratory visualization of three-dimensional thin fiber data. A critical task for the visual analysis of these data is understanding patterns in fiber orientation throughout a volume.The interface enables a new, fluid style of data exploration and fiber orientation analysis by using props to provide needed passive-haptic feedback, making 3D interactions with these fiber structures more controlled. We also contribute a low-level algorithm for extracting fiber centerlines from volumetric imaging. The system was designed and evaluated with two biophotonic experts who currently use it in their lab. As compared to typical practice within their field, the new visualization system provides a more effective way to examine and understand the 3D bioimaging datasets they collect.


Inhibition of Na+ influx and DNA synthesis in human fibroblasts and neuroblastoma-glioma hybrid cells by amiloride analogs.
Identification of a Na+ influx inhibitor that is significantly more potent than amiloride and devoid of the nonspecific effects of amiloride would be of great value in determining the validity of the hypothesis that mitogen-stimulated Na+ influx acts as a signal for induction of cell proliferation. In this study, we evaluated a number of amiloride analogs for potency of Na+ influx inhibition in human fibroblasts (HSWP). One analog, benzamil, was found to exhibit a 60-fold enhanced potency relative to amiloride. We also assessed the relative efficacies with which amiloride and benzamil inhibit Na+ influx and DNA synthesis in HSWP cells and neuroblastoma-glioma hybrid cells (NG108-15). Concentrations of benzamil required for 50% inhibition (ID50) of Na+ influx and DNA synthesis of HSWP cells are in excellent agreement (15 and 18 microM, respectively), an observation which, on the surface, is supportive of the hypothesis in question. Benzamil also inhibits Na+ influx of NG108-15 cells with an ID50 comparable to that for HSWP cells (18 microM) and suppresses DNA synthesis with a slightly higher ID50 (38 microM). Although the benzamil concentrations needed to inhibit cell growth and Na+ influx are in reasonable agreement, caution should be exercised in interpreting the effects of benzamil on cell growth with respect to the role of Na+ influx as we also observed that an analog of benzamil with a reduced ability to inhibit Na+ influx gave inhibition of DNA synthesis at concentrations which do not inhibit Na+ influx.


A physical interaction network of dengue virus and human proteins.
Dengue virus (DENV), an emerging mosquito-transmitted pathogen capable of causing severe disease in humans, interacts with host cell factors to create a more favorable environment for replication. However, few interactions between DENV and human proteins have been reported to date. To identify DENV-human protein interactions, we used high-throughput yeast two-hybrid assays to screen the 10 DENV proteins against a human liver activation domain library. From 45 DNA-binding domain clones containing either full-length viral genes or partially overlapping gene fragments, we identified 139 interactions between DENV and human proteins, the vast majority of which are novel. These interactions involved 105 human proteins, including six previously implicated in DENV infection and 45 linked to the replication of other viruses. Human proteins with functions related to the complement and coagulation cascade, the centrosome, and the cytoskeleton were enriched among the DENV interaction partners. To determine if the cellular proteins were required for DENV infection, we used small interfering RNAs to inhibit their expression. Six of 12 proteins targeted (CALR, DDX3X, ERC1, GOLGA2, TRIP11, and UBE2I) caused a significant decrease in the replication of a DENV replicon. We further showed that calreticulin colocalized with viral dsRNA and with the viral NS3 and NS5 proteins in DENV-infected cells, consistent with a direct role for calreticulin in DENV replication. Human proteins that interacted with DENV had significantly higher average degree and betweenness than expected by chance, which provides additional support for the hypothesis that viruses preferentially target cellular proteins that occupy central position in the human protein interaction network. This study provides a valuable starting point for additional investigations into the roles of human proteins in DENV infection.


Attitudes Toward Technology and Use of Fall Alert Wearables in Caregiving: Survey Study.
Wearable technology for fall alerts among older adult care recipients is one of the more frequently studied areas of technology, given the concerning consequences of falls among this population. Falls are quite prevalent in later life. While there is a growing amount of literature on older adults' acceptance of technology, less is known about how caregivers' attitudes toward technology can impact care recipients' use of such technology. The objective of our study was to examine associations between caregivers' attitudes toward technology for caregiving and care recipients' use of fall alert wearables. This study examined data collected with an online survey from 626 caregivers for adults 50 years and older. Adapted from the technology acceptance model, a structural equation model tested the following prespecified hypotheses: (1) higher perceived usefulness of technologies for caregiving would predict higher perceived value of and greater interest in technologies for caregiving; (2) higher perceived value of technologies for caregiving would predict greater interest in technologies for caregiving; and (3) greater interest in technologies for caregiving would predict greater use of fall alert wearables among care recipients. Additionally, we included demographic factors (eg, caregivers' and care recipients' ages) and caregiving context (eg, caregiver type and caregiving situation) as important predictors of care recipients' use of fall alert wearables. Of 626 total respondents, 548 (87.5%) with all valid responses were included in this study. Among care recipients, 28% used fall alert wearables. The final model had a good to fair model fit: a confirmatory factor index of 0.93, a standardized root mean square residual of 0.049, and root mean square error of approximation of 0.066. Caregivers' perceived usefulness of technology was positively associated with their attitudes toward using technology in caregiving (b=.70, P<.001) and interest in using technology for caregiving (b=.22, P=.003). Greater perceived value of using technology in caregiving predicted greater interest in using technology for caregiving (b=.65, P<.001). Greater interest in using technology for caregiving was associated with greater likelihood of care recipients using fall alert wearables (b=.27, P<.001). The caregiver type had the strongest inverse relationship with care recipients' use of fall alert wearables (unpaid vs paid caregiver) (b=-.33, P<.001). This study underscores the importance of caregivers' attitudes in care recipients' technology use for falls management. Raising awareness and improving perception about technologies for caregiving may help caregivers and care recipients adopt and better utilize technologies that can promote independence and enhance safety.


Value-based care in hepatology.
The migration from legacy fee-for-service reimbursement to payments linked to high-value health care is accelerating in the United States because of new legislation and redesign of payments from the Centers for Medicare and Medicaid Services. Because patients with chronic diseases account for substantial use of health care resources, payers and health systems are focusing on maximizing the value of care for these patients. Because chronic liver diseases impose a major health burden worldwide affecting the health and lives of many individuals and families as well as substantial costs for individuals and payers, hepatologists must understand how they can improve their practices. Hepatologists practice a high-intensity cognitive subspecialty, using complex and costly procedures and medications. High-value patient care requires multidisciplinary coordination, labor-intensive support for critically ill patients, and effective chronic disease management. Under current fee-for-service reimbursement, patient values, medical success, and financial success can all be misaligned. Many current attempts to link health outcomes to reimbursement are based on compliance with process measures, with less emphasis on outcomes that matter most to patients, thus slowing transformation to higher-value team-based care. Outcome measures that reflect the entire cycle of care are needed to assist both clinicians and administrators in improving the quality and value of care. A comprehensive set of outcome measures for liver diseases is not currently available. Numerous researchers now are attempting to fill this gap by devising and testing outcome indicators and patient-reported outcomes for the major liver conditions. These indicators will provide tools to implement a value-based approach for patients with chronic liver diseases to compare results and value of care between referral centers, to perform health technology assessment, and to guide decision-making processes for health authorities. This review sets the groundwork for implementing a value-based, patient-centered approach to chronic liver diseases within a health system. (Hepatology 2017;65:1749-1755).


Pore-scale dispersion in electrokinetic flow through a random sphere packing.
The three-dimensional velocity field and corresponding hydrodynamic dispersion in electrokinetic flow through a random bulk packing of impermeable, nonconducting spheres are studied by quantitative numerical analysis. First, a fixed bed with interparticle porosity of 0.38 is generated using a parallel collective-rearrangement algorithm. Then, the interparticle velocity field is calculated using the lattice-Boltzmann (LB) method, and a random-walk particle-tracking method is finally employed to model advection-diffusion of an inert tracer in the LB velocity field. We demonstrate that the pore-scale velocity profile for electroosmotic flow (EOF) is nonuniform even under most ideal conditions, including a negligible thickness of the electrical double layer compared to the mean pore size, a uniform distribution of the electrokinetic potential at the solid-liquid interface, and the absence of applied pressure gradients. This EOF dynamics is caused by a nonuniform distribution of the local electrical field strength in the sphere packing and engenders significant hydrodynamic dispersion compared to pluglike EOF through a single straight channel. Both transient and asymptotic dispersion behaviors are analyzed for EOF in the context of packing microstructure and are compared to pressure-driven flow in dependence of the average velocity through the bed. A better hydrodynamic performance of EOF originates in a still much smaller amplitude of velocity fluctuations on a mesoscopic scale (covering several particle diameters), as well as on the microscopic scale of an individual pore.


Leveraging the principles of osteopathic medicine to improve diabetes outcomes within a new era of health care reform.
First introduced conceptually decades before the passage of the Patient Protection and Affordable Care Act, the patient-centered medical home (PCMH) has evolved as a foundational element within the larger health care system or medical neighborhood, highlighting a coordinated and comprehensive disease management approach centered on intensive primary care interventions. More recently, in the wake of health care reform, accountable care organizations (ACOs) have been established to help health plans, physicians, hospitals, home health care practitioners, and other health care providers better coordinate care through an incentive-based payment arrangement. Osteopathic medicine is poised to proactively capitalize on these emerging health care models, with the anticipated end result of improved quality of care and reduced health care costs. As such, osteopathic physicians involved in the prevention and care of patients with type 2 diabetes mellitus need to identify models, best practices, and solutions to advance the medical neighborhood.


Pleomorphic adenoma of the parotid.
Pleomorphic adenoma of the parotid is the most common tumor of salivary gland origin, accounting for 60 to 70 percent of all benign salivary gland tumors. This lesion usually presents as a slow-growing painless mass inferior to the pinna of the ear. The diagnosis is based on clinical presentation, magnetic resonance imaging or computed tomography, and fine-needle aspiration biopsy. The treatment is wide excision in which the entire capsule is removed but the facial nerve is spared. Proper diagnosis and treatment are necessary to prevent the complications of tumor recurrence and malignant transformation. Carcinoma expleomorphic adenoma arises in longstanding tumors and has a five-year recurrence rate of 75 percent.


A computer vision-based system for monitoring Vojta therapy.
A neurological illness is t he disorder in human nervous system that can result in various diseases including the motor disabilities. Neurological disorders may affect the motor neurons, which are associated with skeletal muscles and control the body movement. Consequently, they introduce some diseases in the human e.g. cerebral palsy, spinal scoliosis, peripheral paralysis of arms/legs, hip joint dysplasia and various myopathies. Vojta therapy is considered a useful technique to treat the motor disabilities. In Vojta therapy, a specific stimulation is given to the patient's body to perform certain reflexive pattern movements which the patient is unable to perform in a normal manner. The repetition of stimulation ultimately brings forth the previously blocked connections between the spinal cord and the brain. After few therapy sessions, the patient can perform these movements without external stimulation. In this paper, we propose a computer vision-based system to monitor the correct movements of the patient during the therapy treatment using the RGBD data. The proposed framework works in three steps. In the first step, patient's body is automatically detected and segmented and two novel techniques are proposed for this purpose. In the second step, a multi-dimensional feature vector is computed to define various movements of patient's body during the therapy. In the final step, a multi-class support vector machine is used to classify these movements. The experimental evaluation carried out on the large captured dataset shows that the proposed system is highly useful in monitoring the patient's body movements during Vojta therapy.


Tumor-associated macrophages in clear cell renal cell carcinoma express both gastrin-releasing peptide and its receptor: a possible modulatory role of immune effectors cells.
Renal cell carcinomas (RCC) frequently express the gastrin-releasing peptide receptor (GRP-R). Gastrin-releasing peptide (GRP) stimulates tumor cell proliferation and neoangiogenesis. Tumor-associated macrophages (TAM) comprise an important cellular component of these tumors. We analyzed the GRP/GRP-R network in clear cell RCC (ccRCC) and non-clear cell RCC (non-ccRCC) with special regard to its expression by macrophages, tumor cells and microvessels. Gastrin-releasing peptide and GRP-R expression in 17 ccRCC and 9 non-ccRCC were analyzed by RT-PCR, immunohistochemistry and double immunofluorescence staining. Tumor-associated macrophages expressed GRP and GRP receptor in ccRCC. Tumor cells and microvessels showed low to intermediate GRP-R expression in nearly all cases. In 12 ccRCC tumor epithelia also expressed low levels of GRP. Microvascular GRP expression was found in nine cases of ccRCC. For non-RCC, the expression of GRP and GRP receptor expression pattern was similar. Tumor-associated macrophages are the main source of GRP in RCC. GRP receptor on TAM, tumor epithelia and microvessels might be a molecular base of a GRP/GRP receptor network, potentially acting as a paracrine/autocrine modulator of TAM recruitment, tumor growth and neoangiogenesis.


Pro- and Anti-inflammatory Effects of High Cholesterol Diet on Aged Brain.
Both hypercholesterolemia and aging are related to cognitive decline or Alzheimer's disease. However, their interactive influence on the neurodegenerative progress remains unclear. To address this issue, 6-month-old and 16-month-old female mice were fed a 3% cholesterol diet for 8 weeks, followed by hippocampus-related functional, pathological, biochemical and molecular analyses. The high cholesterol diet did not exacerbate age-dependent cognitive decline and hippocampal neuronal death, and even greatly mitigated decreases of synaptophysin and growth associated protein 43 expression in the hippocampus of aged mice. Compared with young controls, aged mice fed normal diet showed mild activation of hippocampal microglia with increased expression of CD68, a marker of the microglial M1 phenotype, and decreased expression of CD206, a marker of the microglial M2 phenotype. More interestingly, the high cholesterol diet not only improved NLRP3 inflammasome activation and IL-1β expression, but also increased levels of anti-inflammatory cytokines IL-4 and IL-6 in the hippocampus of old mice, suggesting playing pro- and anti-neuroinflammatory effects. In addition, the cholesterol rich diet resulted in a defect of the blood-brain barrier of aged hippocampus, as revealed by increased brain albumin content. These results have revealed both harmful and protective effects of high cholesterol diet on aged brain, which helps us to understand that hypercholesterolemia in the aged population is not associated with dementia and cognitive impairment.


Are chronic musculoskeletal pain and generalized joint hypermobility disabling contributors to physical functioning?
Chronic musculoskeletal pain (CMP), Generalized Joint Hypermobility (GJH) and pain-related fear have influence on physical functioning in adolescents. to evaluate differences in physical functioning between adolescents with CMP, GJH or the combination of both, and in addition evaluate the potential contribution of pain-related fear. The design of this study was observational and cross-sectional. The adolescents with CMP were recruited by a physician in rehabilitation medicine and measured in the university outpatient rehabilitation clinic (Adelante/Maastricht University Medical Center+, the Netherlands). The adolescents without CMP were recruited in the Southern area of the Netherlands and measured in the university outpatient rehabilitation clinic (Adelante/Maastricht University Medical Center+, the Netherlands). Four subgroups of adolescents were included; 21 adolescents with CMP without GJH, 9 adolescents with CMP and GJH, 51 adolescents without CMP without GJH, and 11 adolescents without CMP with GJH. Outcome measures were muscle strength and endurance, motor performance, physical activity level, and pain-related fear. Hierarchical regression analyses were used to study differences in physical functioning and the contribution of pain-related fear in adolescents with/without CMP as well as with/without GJH. Adolescents with CMP had decreased muscle strength (P=0.01), endurance (P=0.02), and lower motor performance (P<0.01) compared to adolescents without CMP. Higher levels of pain-related fear were related to decreased muscle strength (P=0.01), endurance (P<0.01), and motor performance (P<0.01). No differences in physical functioning and pain-related fear between hypermobile and non-hypermobile adolescents with CMP were found. Adolescents with CMP had decreased muscle strength and motor performance associated with increased levels of pain-related fear compared to adolescents without CMP. The association of being hypermobile with physical functioning is not more pronounced in adolescents with CMP. No differences were found in physical functioning and pain-related fear between hypermobile adolescents with CMP compared to non-hypermobile adolescents with CMP. Future rehabilitation treatment in hypermobile adolescents with CMP should also focus on psychological components, such as pain-related fear.


Coherence analysis of trunk and leg acceleration reveals altered postural sway strategy during standing in persons with multiple sclerosis.
Balance task performance is affected in persons with multiple sclerosis (PwMS), but the control strategies used to perform specific tasks are not well understood. The purpose of this study was to evaluate segmental control during quiet standing in PwMS and controls to understand whether MS alters use of the ankle and hip strategies to manage postural sway. Coherence of acceleration between the trunk and legs was evaluated with accelerometers placed on the sacrum and lower leg. Thirty-six PwMS and 20 healthy control subjects performed quiet standing with eyes open and closed while center of pressure (CoP) and acceleration of postural sway was measured. Acceleration frequencies were divided into lower frequencies (≤1.0 Hz) and higher frequencies (>1.0 Hz) to categorize sway characteristics. With eyes open, coherence was significantly lower in PwMS compared to controls at lower frequencies only. With eyes closed, coherence was significantly lower in PwMS compared to controls, who use an ankle strategy at lower frequencies only, at both lower and higher frequencies. Both groups showed decreased coherence with increasing frequency when eyes were open and closed. Coherence was significantly correlated with CoP sway area in PwMS during the eyes closed condition only. The reduced coherence in PwMS during both lower and higher frequency sway indicates PwMS utilize a mixed ankle-hip sway strategy regardless of sway frequency. This is in contrast to sway in healthy subjects which utilizes an ankle strategy at lower frequencies and a mixed strategy at higher frequencies. Lack of adaptability in segmental control strategy likely contributes to abnormal postural control, as reflected by CoP sway patterns, in PwMS.


Subcellular compartmentalization of PKM2 identifies anti-PKM2 therapy response in vitro and in vivo mouse model of human non-small-cell lung cancer.
Pyruvate kinase M2 (PKM2) is an alternatively spliced variant, which mediates the conversion of glucose to lactate in cancer cells under normoxic conditions, known as the Warburg effect. Previously, we demonstrated that PKM2 is one of 97 genes that are overexpressed in non-small-cell lung cancer (NSCLC) cell lines. Herein, we demonstrate a novel role of subcellular PKM2 expression as a biomarker of therapeutic response after targeting this gene by shRNA or small molecule inhibitor (SMI) of PKM2 enzyme activity in vitro and in vivo. We examined two established lung cancer cell lines, nine patients derived NSCLC and three normal lung fibroblast cell lines for PKM2 mRNA, protein and enzyme activity by RT-qPCR, immunocytochemistry (ICC), and Western blot analysis. All eleven NSCLC cell lines showed upregulated PKM2 enzymatic activity and protein expression mainly in their cytoplasm. Targeting PKM2 by shRNA or SMI, NSCLC cells showed significantly reduced mRNA, enzyme activity, cell viability, and colony formation, which also downregulated cytosolic PKM2 and upregulated nuclear enzyme activities. Normal lung fibroblast cell lines did not express PKM2, which served as negative controls. PKM2 targeting by SMI slowed tumor growth while gene-silencing significantly reduced growth of human NSCLC xenografts. Tumor sections from responding mice showed >70% reduction in cytoplasmic PKM2 with low or undetectable nuclear staining by immunohistochemistry (IHC). In sharp contrast, non-responding tumors showed a >38% increase in PKM2 nuclear staining with low or undetectable cytoplasmic staining. In conclusion, these results confirmed PKM2 as a target for cancer therapy and an unique function of subcellular PKM2, which may characterize therapeutic response to anti-PKM2 therapy in NSCLC.


Phase Ib Results of the Rational Combination of Selumetinib and Cyclosporin A in Advanced Solid Tumors with an Expansion Cohort in Metastatic Colorectal Cancer.
MEK inhibition is of interest in cancer drug development, but clinical activity in metastatic colorectal cancer (mCRC) has been limited. Preclinical studies demonstrated Wnt pathway overexpression in KRAS-mutant cell lines resistant to the MEK inhibitor, selumetinib. The combination of selumetinib and cyclosporin A, a noncanonical Wnt pathway modulator, demonstrated antitumor activity in mCRC patient-derived xenografts. To translate these results, we conducted a NCI Cancer Therapy Evaluation Program-approved multicenter phase I/IB trial (NCT02188264) of the combination of selumetinib and cyclosporin A. Patients with advanced solid malignancies were treated with the combination of oral selumetinib and cyclosporin A in the dose escalation phase, followed by an expansion cohort of irinotecan and oxaliplatin-refractory mCRC. The expansion cohort utilized a single-agent selumetinib "run-in" to evaluate FZD2 biomarker upregulation and KRAS-WT and KRAS-MT stratification to identify any potential predictors of efficacy. Twenty and 19 patients were enrolled in dose escalation and expansion phases, respectively. The most common adverse events and grade 3/4 toxicities were rash, hypertension, and edema. Three dose-limiting toxicities (grade 3 hypertension, rash, and increased creatinine) were reported. The MTD was selumetinib 75 mg twice daily and cyclosporin A 2 mg/kg twice daily on a 28-day cycle. KRAS stratification did not identify any differences in response between KRAS-WT and KRAS-MT cancers. Two partial responses, 18 stable disease, and 10 progressive disease responses were observed. Combination selumetinib and cyclosporin A is well tolerated, with evidence of activity in mCRC. Future strategies for concept development include identifying better predictors of efficacy and improved Wnt pathway modulation.Significance: These findings translate preclinical studies combining selumetinib and cyclosporin into a phase I first-in-human clinical trial of such a combination in patients with advanced solid malignancies. Cancer Res; 78(18); 5398-407. ©2018 AACR.


Different modes of growth hormone (GH) administration do not change GH binding protein activity in man.
Studies in rodents have shown GH binding protein (GHBP) levels to be dependent on the mode of GH administration. The aim is to determine whether GHBP levels in man are also modulated by the pattern of GH administration. Six GH deficient subjects participated in a randomized study in which 2 IU GH were administered either as (i) a continuous 24-hour infusion, (ii) two intravenous boluses or (iii) eight intravenous boluses every 3 hours. In a second study, six normal men received a single subcutaneous injection of 0.2 U/kg GH and GHBP activity was measured over 24 hours. Control data were obtained from an untreated group of six age-matched normal men. GHBP activity was measured by immunoprecipitation using a monoclonal antibody that recognizes the human GHBP, and expressed as percentage specific binding of 125I-GH in 50 microliters of serum. GHBP activity was not significantly different between the GH deficient and normal subjects. GHBP activity did not rise significantly during GH administration with each of the three intravenous patterns of delivery nor were there any significant differences between treatments. In the second study, GHBP activity did not change significantly following subcutaneous GH injection nor did results differ from untreated normal controls. The level of GHBP in man is not dependent on GH secretory status or altered by short-term GH treatment or the mode of administration. These findings stand in contrast to GH treatment effects in rodents and suggest that GH regulation of GHBP may be different between species.


Exercise-induced bronchoconstriction: pathogenesis.
There is still active debate on the acute mechanism of exercise-induced bronchoconstriction (EIB). Although it is unlikely that vasoconstriction and hyperemia of the bronchial vasculature are essential events for EIB, it is likely that this vasculature enhances the airway response to dehydration and contributes to the pathogenesis of EIB, particularly in elite athletes. Accumulating evidence suggests that airway smooth muscle (ASM) becomes more sensitive as a result of repeated exposure to bulk plasma in response to airway injury from dehydration. Recent evidence also demonstrates sufficient concentrations of mediators that could affect ASM. Paradoxically, mediator release from mast cells may be enhanced and their contractile effects greater when beta(2)-receptor agonists are taken daily. The effect of drugs that have the potential to reduce microvascular leak and reduce or inhibit release or action of these mediators needs to be investigated in elite athletes.


How homosexual men believe they became infected with HIV: the role of risk-reduction behaviors.
A range of HIV risk reduction strategies has been described in homosexual men who practice unprotected anal intercourse (UAI), including serosorting, strategic positioning, and negotiating around an HIV-positive partner's viral load. It is uncertain how commonly these behaviors might result in HIV infection. We describe sexual behaviors and associated risk reduction strategies reported by homosexual men in their accounts of their recent seroconversion. Homosexual men recently diagnosed with primary HIV infection between 2003 and 2006 were invited to participate in a nurse-administered survey. Among 158 men enrolled, 143 (91%) were able to identify the high-risk event that they believed led to their HIV seroconversion, and this involved UAI in 102 (71%). Among these 102 men, 21 (21%) reported they were certain that the source partner was HIV-negative. Ten men (10%) reported insertive UAI as the highest risk behavior. Of the 21 men who reported knowing the HIV-positive partner's viral load, 9 reported that the man had an undetectable viral load (43%). In 38% of the high-risk events involving UAI, infection occurred when the reported risk event involved serosorting, strategic positioning, or intercourse with a man whose viral load was believed to be undetectable.


The ATM-related cofactor Tra1 is a component of the purified SAGA complex.
The SAGA histone acetyltransferase/transcriptional adaptor complex is composed of multiple transcriptional regulators including Ada, Spt, and TAFII proteins. Here we identify an additional novel subunit of the complex, Tra1, an ATM/PI-3-kinase-related homolog of the human TRRAP cofactor, which is essential for c-Myc and E2F-mediated oncogenic transformation. Mass spectrometry, immunoblotting, and immunoprecipitation experiments confirm the stable association of this protein within SAGA. In addition, the Tra1 protein is a component of at least two other histone acetyltransferase protein complexes. These results indicate a role for Tra1 in the regulation of transcriptional activation through the recruitment of HAT activity to an activator-bound promoter.


Human growth hormone: complementary DNA cloning and expression in bacteria.
The nucleotide sequence of a DNA complementary to human growth hormone messenger RNA was cloned; it contains 29 nucleotides in its 5' untranslated region, the 651 nucleotides coding for the prehormone, and the entire 3' untranslated region (108 nucleotides). The data reported predict the previously unknown sequence of the signal peptide of human growth hormone and, by comparison with the previously determined sequences of rat growth hormone and human chorionic somatomammotropin, strengthens the hypothesis that these genes evolved by gene duplication from a common ancestral sequence. The human growth hormone gene sequences have been linked in phase to a fragment of the trp D gene of Escherichia coli in a plasmid vehicle, and a fusion protein is synthesized at high level (approximately 3 percent of bacterial protein) under the control of the regulatory region of the trp operon. This fusion protein (70 percent of whose amino acids are coded for by the human growth hormone gene) reacts specifically with antibodies to human growth hormone and is stable in E. coli.


Genetic interactions between HOP1, RED1 and MEK1 suggest that MEK1 regulates assembly of axial element components during meiosis in the yeast Saccharomyces cerevisiae.
During meiosis, axial elements are generated by the condensation of sister chromatids along a protein core as precursors to the formation of the synaptonemal complex (SC). Functional axial elements are essential for wild-type levels of recombination and proper reductional segregation at meiosis I. Genetic and cytological data suggest that three meiosis-specific genes, HOP1, RED1 and MEK1, are involved in axial element formation in the yeast Saccharomyces cerevisiae. HOP1 and RED1 encode structural components of axial elements while MEK1 encodes a putative protein kinase. Using a partially functional allele of MEK1, new genetic interactions have been found between HOP1, RED1 and MEK1. Overexpression of HOP1 partially suppresses the spore inviability and recombination defects of mek1-974; in contrast, overexpression of RED1 exacerbates the mek1-974 spore inviability. Co-overexpression of HOP1 and RED1 in mek1-974 diploids alleviates the negative effect of overexpressing RED1 alone. Red1p/Red1p as well as Hop1p/Red1p interactions have been reconstituted in two hybrid experiments. Our results suggest a model whereby Mek1 kinase activity controls axial element assembly by regulating the affinity with which Hop1p and Red1p interact with each other.


Beliefs about physical activity and sedentary behaviors of adults with visual impairments.
When exploring reasons why individuals with visual impairments (VI) may or may not engage in physical activity (PA) or sedentary behaviors (SB), theoretically grounded research on the determinants of these behaviors is scarce. Situated in the theory of planned behavior (TpB), the purposes of this study were to: (a) develop a theoretically-sound scale, Beliefs about Physical and Sedentary Behaviors-Visual Impairment (BAPS-VI), to determine if the constructs of TpB are useful in predicting PA and SBs of adults with VI; (b) analyze their beliefs about PA and SBs; and (c) determine which TpB constructs are the best predictors of PA behaviors. Data were collected from adults with VI (n = 209, 65.5% women) using on-line survey methodology. Following reliability estimation, the PA and SB items were reduced using separate Principal Components analyses to examine the underlying dimension of the BAPS-VI in relation to TpB. A hierarchical regression model was used to determine what factors predicted self-reports of minutes of PA. Results supported the theoretical framework of the measure and explained 75% of the variance for intention to engage in PA and SB items, respectively. Six new variables, intention, and demographic data were then regressed on physical activity scores with only intention to engage in PA (β = 0.30, p < 0.01) remaining as a significant predictor of physical activity. The development of the theoretically driven measure and results of this study can inform future research focusing on exploring PA and SBs of adults with VI.


HuR as a negative posttranscriptional modulator in inflammation.
HuR is an RNA binding protein with an alleged role in the posttranscriptional activation of inflammatory mRNAs bearing AU-rich elements (AREs). Here, we show that the inducible increase of HuR in murine innate compartments suppresses inflammatory responses in vivo. In macrophages, HuR overexpression induced the translational silencing of specific cytokine mRNAs despite positive or nominal effects on their corresponding turnover. By using a model system of ARE dysfunction, we demonstrate that HuR does not alter the accumulation of target mRNAs in the absence of the destabilizing functions of Tristetraprolin but synergizes with the translational silencer TIA-1 to reduce the translation of cytokine mRNAs. Our data suggest that HuR acts in a pleiotropic fashion in inflammation through its functional interactions with specific mRNA subsets and negative posttranscriptional modules.


Immunodiagnosis of snake bite.
Management of a patient with snake bite is influenced by the nature of the offending snake. Species diagnosis based on the patient's history and physical signs is often unreliable and the possibility of making a species diagnosis by immunological means has therefore been investigated. Wound aspirates, blister fluids, sera, and urine samples from patients with snake bite were examined for the presence of species-specific venoms using immunodiffusion. A positive species diagnosis was made in 40 out of 101 patients. Immunodiagnosis was especially successful in patients bitten by the puff adder, Bitis arietans, and the African spitting cobra, Naja nigricollis. A higher success rate could probably be achieved using more specific antisera and more sensitive assay techniques.


Effect of Antithrombotic Therapy and Long Endoscopic Submucosal Dissection Procedure Time on Early and Delayed Postoperative Bleeding.
Recent updated guidelines of the Japanese Society of Gastroenterology recommend the use of a single dose of antiplatelet agents in patients undergoing endoscopic submucosal dissection (ESD). However, the postoperative bleeding risk after gastric ESD associated with the continuation or interruption of antithrombotic therapy remains controversial. We aimed to evaluate whether certain factors including interrupted antithrombotic therapy could affect early and delayed post-ESD bleeding risk. Three hundred sixty-four patients with gastric neoplasms were treated with ESD at our hospital between October 2005 and December 2012. Seventy-four patients with interrupted antithrombotic therapy were undertaken with ESD. Early and delayed postoperative bleeding patterns were estimated. Various clinical characteristics such as gender, age, tumor location, tumor size, ESD procedure time, platelet count, and comorbidity were evaluated. There was a significant difference (p = 0.042) in the ESD procedure time between the patients with postoperative bleeding and those without it. There was no significant difference in postoperative bleeding between the patients on antithrombotic therapy and not on it. Moreover, interrupted antithrombotic therapy and platelet count were significantly (p = 0.0461 and p = 0.0059, respectively) associated with early postoperative bleeding in multivariate analysis. In addition, in univariate analysis, ESD procedure time was significantly (p = 0.041) associated with delayed postoperative bleeding. Antithrombotic therapy and prolonged ESD procedure time were significantly associated with early and delayed postoperative bleeding, respectively.


Correlation of higher preoperative American Society of Anesthesiology grade and increased morbidity and mortality rates in patients undergoing spine surgery.
Patients with varied medical comorbidities often present with spinal pathology for which operative intervention is potentially indicated, but few studies have examined risk stratification in determining morbidity and mortality rates associated with the operative treatment of spinal disorders. This study provides an analysis of morbidity and mortality data associated with 22,857 cases reported in the multicenter, multisurgeon Scoliosis Research Society Morbidity and Mortality database stratified by American Society of Anesthesiologists (ASA) physical status classification, a commonly used system to describe preoperative physical status and to predict operative morbidity. The Scoliosis Research Society Morbidity and Mortality database was queried for the year 2007, the year in which ASA data were collected. Inclusion criterion was a reported ASA grade. Cases were categorized by operation type and disease process. Details on the surgical approach and type of instrumentation were recorded. Major perioperative complications and deaths were evaluated. Two large subgroups--patients with adult degenerative lumbar disease and patients with major deformity--were also analyzed separately. Statistical analyses were performed with the chi-square test. The population studied comprised 22,857 patients. Spinal disease included degenerative disease (9409 cases), scoliosis (6782 cases), spondylolisthesis (2144 cases), trauma (1314 cases), kyphosis (831 cases), and other (2377 cases). The overall complication rate was 8.4%. Complication rates for ASA Grades 1 through 5 were 5.4%, 9.0%, 14.4%, 20.3%, and 50.0%, respectively (p = 0.001). In patients undergoing surgery for degenerative lumbar diseases and major adult deformity, similarly increasing rates of morbidity were found in higher-grade patients. The mortality rate was also higher in higher-grade patients. The incidence of major complications, including wound infections, hematomas, respiratory problems, and thromboembolic events, was also greater in patients with higher ASA grades. Patients with higher ASA grades undergoing spinal surgery had significantly higher rates of morbidity than those with lower ASA grades. Given the common application of the ASA system to surgical patients, this grade may prove helpful for surgical decision making and preoperative counseling with regard to risks of morbidity and mortality.


Epidemiologic assessment of mortality, building collapse pattern, and medical response after the 1992 earthquake in Turkey. Disaster Reanimatology Study Group (DRSG).
Post-earthquake engineering and epidemiologic assessments are important for the development of injury prevention strategies. This paper describes mortality and its relationship to building collapse patterns and initial medical responses following the 1992 earthquake in Erzincan, Turkey. The study consisted of: 1) background data collection and review; 2) design and implementation of a field survey; and 3) site inspection of building collapse patterns. The survey included: 1) national (n = 11) and local (n = 17) officials; 2) medical and search and rescue (SAR) workers (n = 38); and 3) a geographically stratified random sample of lay survivors (n = 105). The survey instruments were designed to gather information regarding location, injuries, initial actions and prior training of survivors and responders, and the location, injuries, and management of dead and dying victims. A case-control design was constructed to assess the relationship between mortality, location, and building collapse pattern. There was extensive structural damage throughout the region, especially in the city where mid-rise, unreinforced masonry buildings (MUMBs) incorporating a "soft" first floor design (large store windows for commercial use) and one story adobe structures were most vulnerable to collapse. Of 526 people who died in the city, 87% (n = 456) were indoors at the time of the earthquake. Of these, 92% (n = 418) died in MUMBs. Of 54 witnessed deaths, 55% (n = 28) of victims died slowly, the majority of whom (n = 26) were pinned or trapped (p < 0.05). Of 42 MUMB occupants identified through the survey, those who died (n = 25) were more likely to have been occupying the ground floor when compared with survivors (n = 28) (p < 0.01). Official medical and search and rescue responders arrived after most deaths had occurred. Prior first-aid or rescue training of lay, uninjured survivors was associated with a higher likelihood of rescuing and resuscitating others (p < 0.001). During an earthquake, MUMBs with soft ground floor construction are highly lethal, especially for occupants on the the ground floor, suggesting that this building type is inappropriate for areas of seismic risk. The vulnerability of MUMBs appears due to a lack of lateral force resistance as a result of the use of glass store front windows and the absence of shear walls. The prevalence of this building type in earthquake-prone regions needs to be investigated further. A large portion of victims dying in an earthquake die slowly at the scene of injury. Prior public first-aid and rescue training programs increase participation in rescue efforts in major earthquakes and may improve survival.


Shaping the gradient by nonchemotactic chemokine receptors.
Chemokines are a class of inflammatory mediators which main function is to direct leukocyte migration through the binding to G protein-coupled receptors (GPCRs). In addition to these functional, signal-transducing chemokine receptors other types of receptors belonging to the chemokine GPCR family were identified. They are called atypical or decoy chemokine receptors because they bind and degrade chemokines but do not transduce signals or activate cell migration. Here there is the summary of two recent papers that identified other nonchemotactic chemokine receptors: the Duffy antigen receptor for chemokines (DARC) that mediates trancytosis of chemokines from tissue to vascular lumen promoting chemokine-mediated leukocyte transmigration and chemokine (CC motif) receptor-like 2 (CCRL2) that neither internalizes its ligands nor transduces signals but presents bound ligands to functional signaling receptors improving their activity. Collectively these nonchemotactic chemokine receptors do not directly induce cell migration, but appear nonetheless to play a nonredundant role in leukocyte recruitment by shaping the chemoattractant gradient, either by removing, transporting or concentrating their cognate ligands.


Lesion detection by [89Zr]Zr-DFO-girentuximab and [18F]FDG-PET/CT in patients with newly diagnosed metastatic renal cell carcinoma.
The main objective of this preliminary analysis of the IMaging PAtients for Cancer drug selecTion (IMPACT)-renal cell cancer (RCC) study is to evaluate the lesion detection of baseline contrast-enhanced CT, [89Zr]Zr-DFO-girentuximab-PET/CT and [18F]FDG-PET/CT in detecting ccRCC lesions in patients with a good or intermediate prognosis metastatic clear cell renal cell carcinoma (mccRCC) according to the International Metastatic Database Consortium (IMDC) risk model. Between February 2015 and March 2018, 42 newly diagnosed mccRCC patients with good or intermediate prognosis, eligible for watchful waiting, were included. Patients underwent CT, [89Zr]Zr-DFO-girentuximab-PET/CT and [18F]FDG-PET/CT at baseline. Scans were independently reviewed and lesions of ≥10 mm and lymph nodes of ≥15 mm at CT were analyzed. For lesions with [89Zr]Zr-DFO-girentuximab or [18F]FDG-uptake visually exceeding background uptake, maximum standardized uptake values (SUVmax) were measured. A total of 449 lesions were detected by ≥1 modality (median per patient: 7; ICR 4.25-12.75) of which 42% were in lung, 22% in lymph nodes and 10% in bone. Combined [89Zr]Zr-DFO-girentuximab-PET/CT and CT detected more lesions than CT alone: 91% (95%CI: 87-94) versus 56% (95%CI: 50-62, p = 0.001), respectively, and more than CT and [18F]FDG-PET/CT combined (84% (95%CI:79-88, p < 0.005). Both PET/CTs detected more bone and soft tissue lesions compared to CT alone. The addition of [89Zr]Zr-DFO-girentuximab-PET/CT and [18F]FDG-PET/CT to CT increases lesion detection compared to CT alone in newly diagnosed good and intermediate prognosis mccRCC patients eligible for watchful waiting.


Topical antiseptics for chronic suppurative otitis media.
Chronic suppurative otitis media (CSOM), sometimes referred to as chronic otitis media (COM), is a chronic inflammation and infection of the middle ear and mastoid cavity, characterised by ear discharge (otorrhoea) through a perforated tympanic membrane. The predominant symptoms of CSOM are ear discharge and hearing loss. Topical antiseptics, one of the possible treatments for CSOM, inhibit the micro-organisms that may be responsible for the infection. Antiseptics can be used alone or in addition to other treatments for CSOM, such as antibiotics or ear cleaning (aural toileting). Antiseptics or their application can cause irritation of the skin of the outer ear, manifesting as discomfort, pain or itching. Some antiseptics (such as alcohol) may have the potential to be toxic to the inner ear (ototoxicity), with a possible increased risk of causing sensorineural hearing loss, dizziness or tinnitus. To assess the effects of topical antiseptics for people with chronic suppurative otitis media. The Cochrane ENT Information Specialist searched the Cochrane ENT Register; Central Register of Controlled Trials (CENTRAL; 2019, Issue 4, via the Cochrane Register of Studies); Ovid MEDLINE; Ovid Embase; CINAHL; Web of Science; ClinicalTrials.gov; ICTRP and additional sources for published and unpublished trials. The date of the search was 1 April 2019. We included randomised controlled trials (RCTs) with at least a one-week follow-up involving patients (adults and children) who had chronic ear discharge of unknown cause or CSOM, where the ear discharge had continued for more than two weeks. The interventions were any single, or combination of, topical antiseptic agent of any class, applied directly into the ear canal as ear drops, powders or irrigations, or as part of an aural toileting procedure. Two main comparisons were topical antiseptics compared to: a) placebo or no intervention; and b) another topical antiseptic (e.g. topical antiseptic A versus topical antiseptic B). Within each comparison we separated studies where both groups of patients had received topical antiseptics a) alone or with aural toileting and b) on top of antibiotic treatment. We used the standard Cochrane methodological procedures. We used GRADE to assess the certainty of the evidence for each outcome. Our primary outcomes were: resolution of ear discharge or 'dry ear' (whether otoscopically confirmed or not), measured at between one week and up to two weeks, two weeks to up to four weeks, and after four weeks; health-related quality of life using a validated instrument; ear pain (otalgia) or discomfort or local irritation. Secondary outcomes included hearing, serious complications and ototoxicity measured in several ways. Five studies were included. It was not possible to calculate the total number of participants as two studies only provided the number of ears included in the study. A. Topical antiseptic (boric acid) versus placebo or no treatment (all patients had aural toileting) Three studies compared topical antiseptics with no treatment, with one study reporting results we could use (254 children; cluster-RCT). This compared the instillation of boric acid in alcohol drops versus no ear drops for one month (both arms used daily dry mopping). We made adjustments to the data to account for the intra-cluster correlation. The very low certainty of the evidence means it is uncertain whether or not treatment with an antiseptic leads to an increase in resolution of ear discharge at both four weeks (risk ratio (RR) 1.94, 95% confidence interval (CI) 1.20 to 3.16; 174 participants) and at three to four months (RR 1.73, 95% CI 1.21 to 2.47; 180 participants). This study narratively described no differences in suspected ototoxicity or hearing outcomes between the arms (very low-certainty evidence). None of the studies reported results for health-related quality of life, adverse effects or serious complications. B. Topical antiseptic A versus topical antiseptic B Two studies compared different antiseptics but only one (93 participants), comparing a single instillation of boric acid powder with daily acetic acid ear drops, provided any information for this comparison. The very low certainty of the evidence means that it is uncertain whether more patients had resolution of ear discharge with boric acid powder compared to acetic acid at four weeks (RR 2.61, 95% CI 1.51 to 4.53; 93 participants), or whether there was a difference between the arms with respect to ear discomfort due to the low number of reported events (RR 0.10, 95% CI 0.01 to 1.81; 93 participants). Narratively, the study reported no difference in hearing outcomes between the groups. None of the included studies reported any of the other primary or secondary outcomes. Due to paucity of the evidence and the very low certainty of that which is available the effectiveness and safety profile of antiseptics in the treatment of CSOM is uncertain.


Compliance with guidelines on antibiotic prophylaxis in hip and knee arthroplasty in Italy: results of the GISIO-ISChIA project.
The Perioperative Antibiotic Prophylaxis (PAP) contributes considerably to the total amount of antibiotics used in hospitals and has been shown to be associated with increase in antibiotic resistance and healthcare costs. The level of compliance with the national guidelines of current practices of PAP for elective hip and knee prosthesis procedures in a network of Italian hospitals (the multicentre Surgical Site Infection surveillance project GISIO-ISChIA), has been evaluated. Compliance of the current prophylactic antibiotic practices with the published national guidelines was assessed for each included operative procedure, considering indication, timing of administration, prescribed antimicrobial agent and total duration of prophylaxis. A total of 14 hospitals and 1285 surgical procedures were included. 99.4% of patients received antimicrobial prophylaxis pre-operatively and 73.0% of patients received PAP within the recommended time period (within 60 minutes before incision). The rate of compliance with discontinuation of PAP within 24 hours after initiation of surgery was 70.2%. Taking into account all doses administered, the antibiotic was chosen appropriately in 57.7% of patients. PAP was performed appropriately, in accordance with national guidelines, in 43.6% of surgical operations. Given the increasing number of replacement procedures in Italy and in Europe, the gap between the evidence-based guidelines and practice must be appropriately addressed in order to improve PAP practices.


Phylogenetics of fanged frogs: testing biogeographical hypotheses at the interface of the asian and Australian faunal zones.
The interface of the Asian and Australian faunal zones is defined by a network of deep ocean trenches that separate intervening islands of the Philippines and Wallacea (Sulawesi, the Lesser Sundas, and the Moluccas). Studies of this region by Wallace marked the genesis of the field of biogeography, yet few workers have used molecular methods to investigate the biogeography of taxa whose distribution spans this interface. Some taxa, such as the fanged frogs of the ranid genus Limnonectes, have distributions on either side of the zoogeographical lines of Wallace and Huxley, offering an opportunity to ask how frequently these purported barriers were crossed and by what paths. To examine diversification of Limnonectes in Southeast Asia, the Philippines, and Wallacea, we estimated a phylogeny from mitochondrial DNA sequences obtained from a robust geographic sample. Our analyses suggest that these frogs dispersed from Borneo to the Philippines at least twice, from Borneo to Sulawesi once or twice, from Sulawesi to the Philippines once, and from the Philippines to Sulawesi once. Dispersal to the Moluccas occurred from Sulawesi and to the Lesser Sundas from Java/Bali. Species distributions are generally concordant with Pleistocene aggregate island complexes of the Philippines and with areas of endemism on Sulawesi. We conclude that the recognition of zoogeographic lines, though insightful, may oversimplify the biogeography of widespread taxa in this region.


Active surveillance for prostate cancer: a systematic review of contemporary worldwide practices.
In the last decade, active surveillance (AS) has emerged as an acceptable choice for low-risk prostate cancer (PC), however there is discordance amongst large AS cohort studies with respect to entry and monitoring protocols. We systematically reviewed worldwide AS practices in studies reporting ≥5 years follow-up. We searched PubMed and Medline 2000-now and identified 13 AS cohorts. Three key areas were identified: (I) patient selection; (II) monitoring protocols; (III) triggers for intervention-(I) all studies defined clinically localised PC diagnosis as T2b disease or less and most agreed on prostate-specific antigen (PSA) threshold (<10 µg/L) and Gleason score threshold (3+3). Inconsistency was most notable regarding pathologic factors (e.g., number of positive cores); (II) all agreed on PSA surveillance as crucial for monitoring, and most agreed that confirmatory biopsy was required within 12 months of initiation. No consensus was reached on optimal timing of digital rectal examination (DRE), general health assessment or re-biopsy strategies thereafter; (III) there was no universal agreement for intervention triggers, although Gleason score, number or percentage of positive cancer cores, maximum cancer length (MCL) and PSA doubling time were used by several studies. Some also used imaging or re-biopsy. Despite consistent high progression-free/cancer-free survival and conversion-to-treatment rates, heterogeneity exists amongst these large AS cohorts. Combining existing evidence and gathering more long-term evidence [e.g., the Movember's Global AS database or additional information on use of magnetic resonance imaging (MRI)] is needed to derive a broadly supported guideline to reduce variation in clinical practice.


A tri-component knee plug for the 3rd generation of autologous chondrocyte implantation.
Here, we report a newly designed knee plug to be used in the 3rd generation of Autologous Chondrocyte Implantation (ACI) in order to heal the damaged knee cartilage. It is composed of three components: The first component (Bone Portion) is a 3D printed hard scaffold with large pores (~ 850 µm), made by hydroxyapatite and β-tricalcium phosphate to accommodate the bony parts underneath the knee cartilage. It is a cylinder with a diameter of 20 mm and height of 7.5 mm, with a slight dome shape on top. The plug also comprises a Cartilage Portion (component 2) which is a 3D printed gelatin/elastin/sodium-hyaluronate soft thick porous membrane with large pores to accommodate chondrocytes. Cartilage Portion is secured on top of the Bone Portion using mechanical interlocking by designing specific knobs in the 3D printed construct of the Cartilage Portion. The third component of the plug (Film) is a stitchable permeable membrane consisting of polycaprolactone (PCL) on top of the Cartilage Portion to facilitate sliding of the knee joint and to hold the entire plug in place while allowing nutrients delivery to the Cartilage Portion. The PCL Film is prepared using a combination of film casting and sacrificial material leaching with a pore size of 10 µm. It is surface modified to have specific affinity with the Cartilage Portion. The detailed design criteria and production process of this plug is presented in this report. Full in vitro analyses have been performed, which indicate the compatibility of the different components of the plug relative to their expected functions.


Diagnostic segregation of human brain tumours using Fourier-transform infrared and/or Raman spectroscopy coupled with discriminant analysis.
The most common initial treatment received by patients with a brain tumour is surgical removal of the growth. Precise histopathological diagnosis of brain tumours is to some extent subjective. Furthermore, currently available diagnostic imaging techniques to delineate the excision border during cytoreductive surgery lack the required spatial precision to aid surgeons. We set out to determine whether infrared (IR) and/or Raman spectroscopy combined with multivariate analysis could be applied to discriminate between normal brain tissue and different tumour types (meningioma, glioma and brain metastasis) based on the unique spectral "fingerprints" of their biochemical composition. Formalin-fixed paraffin-embedded tissue blocks of normal brain and different brain tumours were de-waxed, mounted on low-E slides and desiccated before being analyzed using attenuated total reflection Fourier-transform IR (ATR-FTIR) and Raman spectroscopy. ATR-FTIR spectroscopy showed a clear segregation between normal and different tumour subtypes. Discrimination of tumour classes was also apparent with Raman spectroscopy. Further analysis of spectral data revealed changes in brain biochemical structure associated with different tumours. Decreased tentatively-assigned lipid-to-protein ratio was associated with increased tumour progression. Alteration in cholesterol esters-to-phenylalanine ratio was evident in grade IV glioma and metastatic tumours. The current study indicates that IR and/or Raman spectroscopy have the potential to provide a novel diagnostic approach in the accurate diagnosis of brain tumours and have potential for application in intra-operative diagnosis.


Psychological Distress, Cognition, and Functional Disability Trajectory Profiles of Aging in Primary Care Older Adults.
To identify profiles of aging by combining psychological distress, cognition and functional disability, and their associated factors. Data were drawn from the Étude sur la Santé des Aînés-Services study and included 1585 older adults. Sociodemographic, psychosocial, lifestyle and health factors were informed from structured interviews. Group-based multi-trajectory modeling and multinomial logistic regression were used to identify aging profiles and correlates. Sampling weights were applied to account for the sampling plan. The weighted sample size was 1591. Three trajectories were identified: a favorable (79.0%), intermediate (14.5%), and severe scenario (6.5%). Factors associated with the severe scenario were older age, male gender, lower education, the presence of anxiety disorders, low physical activity, and smoking. Membership in the intermediate scenario was associated with daily hassles, physical disorders, anxiety and depression, antidepressant/psychotherapy use, low physical activity, and no alcohol use. High social support was protective against less favorable profiles. Symptoms of anxiety and depression and high burden of physical disorders were associated with less favorable trajectories. Modifiable lifestyle factors have a significant effect on healthy aging. Assessment and management of anxio-depressive symptoms are important in older adults. Clinical interventions including access to psychotherapy and promotion of healthier lifestyles should be considered.


Population genetics of tumours.
The available evidence suggests that cancer is essentially a somatic evolutionary process involving a series of mutations. Each mutation gives some advantage to a selected clone, and expansion then occurs within that selected clone. The advantages are associated with both growth rate and factors leading to independent growth. The aim of this paper is first to give some background information on genetic changes in tumours, using colorectal cancer as an example. We will then introduce a mathematical model that explains many phenomena associated with the development of benign tumours and the long lag periods that are characteristic of the development of human tumours. The model addresses populations of cells and not populations of people.


Ultrasound-guided percutaneous lung biopsy in sheep.
Lung biopsies were taken through the ninth intercostal space on the two sides of the thorax of 10 clinically healthy sheep, using a Tru-Cut needle on one side and the Bard Biopty-System on the other. Nine of the 10 sheep remained clinically healthy, but one coughed transiently and had a mild bloody discharge from the right nostril immediately after the biopsy. The sheep were slaughtered 10 days later and the lungs and pleura were examined macroscopically; there were either no lesions or only small scars visible at the sites of the biopsies. However, well-developed subpleural nodules due to parasites were observed in some of the lungs. There were no adhesions between the costal and pulmonary pleurae. Of the 20 biopsy specimens, 18 were ideal for histological examination, and none of them was histologically normal. Mild interstitial pneumonia was diagnosed in 15 specimens, chronic bronchiolitis in nine specimens, and hyperplasia of smooth muscle or connective tissue in 17.


Dynamics of myeloid cell populations during relapse-preventive immunotherapy in acute myeloid leukemia.
Relapse of leukemia in the postchemotherapy phase contributes to the poor prognosis and survival in patients with acute myeloid leukemia (AML). In an international phase IV trial (ClinicalTrials.gov; NCT01347996), 84 patients with AML in first complete remission who had not undergone transplantation received immunotherapy with histamine dihydrochloride (HDC) and low-dose IL-2 with the aim of preventing relapse. The dynamics of myeloid cell counts and expression of activation markers was assessed before and after cycles of immunotherapy and correlated with clinical outcome in terms of relapse risk and survival. During cycles, a pronounced increase in blood eosinophil counts was observed along with a reduction in monocyte and neutrophil counts. A strong reduction of blood monocyte counts during the first HDC/IL-2 treatment cycle predicted leukemia-free survival. The HDC component of the immunotherapy exerts agonist activity at histamine type 2 receptors (H2Rs) that are expressed by myeloid cells. It was observed that the density of H2 R expression in blood monocytes increased during cycles of immunotherapy and that high monocyte H2R expression implied reduced relapse risk and improved overall survival. Several other activation markers, including HLA-DR, CD86, and CD40, were induced in monocytes and dendritic cells during immunotherapy but did not predict clinical outcome. In addition, expression of HLA-ABC increased in all myeloid populations during therapy. A low expression of HLA-ABC was associated with reduced relapse risk. These results suggest that aspects of myeloid cell biology may impact clinical benefit of relapse-preventive immunotherapy in AML.