Frequency channel-dependent selectivity for temporal call characteristics in gray treefrogs, Hyla versicolor.
Sensory receptors transmit information on multiple stimulus dimensions. Much remains to be understood about how the processing of different signal characteristics is partitioned and integrated in different areas of the nervous system. Amphibian hearing involves two morphologically distinct inner-ear organs that process different components of the frequency spectrum. Many anuran signals contain two frequency peaks, each one matching the sensitivity of one of these two organs. We hypothesized that the processing of temporal characteristics of acoustic signals would differ in these two frequency channels, perhaps because of differences in the response properties of the two inner-ear organs. We tested this hypothesis in the gray treefrog, Hyla versicolor; male advertisement calls of this species contain a bimodal frequency spectrum. We generated synthetic male advertisement calls in which we independently manipulated the pattern of amplitude modulation in the low-frequency peak or the high-frequency peak and measured the attractiveness of these stimuli to females in single-speaker and two-speaker phonotaxis tests. We obtained multiple lines of evidence that females were more selective for fine-temporal characteristics in the high-frequency peak. We discuss the potential implications of frequency channel-dependent temporal processing for signal evolution and suggest that additional neurophysiological investigations of the anuran auditory periphery will give important insights into how the nervous system partitions the encoding of multiple characteristics of complex signals.


Determination of fluid leakages in the different screw-retained implant-abutment connections in a mechanical artificial mouth.
This study shows the potential risk of microfiltration between two different types of implant-abutment connections screwed at 45 Ncm: external and internal. For the first time the use of a mechanical artificial mouth is used with the values (compression and torsion loads with a frequency of 2 Hz) of the human chewing. The mechanical tests were performed with an artificial saliva at 37°C. The microgap in the connection was measured by an Image Analysis software incorporated in a high resolution scanning electron microscopy. Implant connections were filled with methylene blue by using self-adjustable precision pipettes and the quantity of leakage was determined by high sensitivity spectometry. We showed that the internal connection has lower microgaps compared to the external ones and these microgaps increased with the number of mechanical cycles. The leakage of methylene blue was higher when the external connection was performed. Microgaps and the influence of the mechanical loads are very important for the long-term behavior avoiding the bacteria colonization in the dental implants. These aspects should be known by the implantologists.


Prevalence and correlates of HIV and hepatitis B virus coinfection in Northern Alberta.
HIV and hepatitis B virus (HBV) share transmission routes, and coinfection is associated with higher morbidity and mortality. To date, no Canadian studies have examined HIV-HBV coinfection. To examine the prevalence and correlates of HIV and HBV coinfections in Northern Alberta. The present study was a retrospective database review of all HIV-infected (HIV+) individuals in Northern Alberta from 1982 to 2010 and a chart review of HBV surface antigen-positive individuals for whom charts were available (46.2%). Of 2844 HIV+ patients, 2579 (90.7%) had been tested for HBV surface antigen, and 143 (5.5%) of these were HBV coinfected. Coinfected males were primarily Caucasian (70.8%), and coinfected females were primarily black (56.4%) or Aboriginal (31.3%). Coinfected individuals were more likely to be male (88.1% versus 71.3%; P<0.001) and to have died (34.3% versus 17.9%; P<0.001). The prevalence of coinfection with HBV in HIV-infected patients in Northern Alberta is lower than reported in other developed nations. The pattern of coinfections in Northern Alberta likely follows immigration trends. Recognition and management may be improving with time; however, further research and additional strategies are required to enhance the prevention, identification and management of HBV infection in HIV-infected individuals.


Interaction between SLC6A4 promoter variants and childhood trauma on the age at onset of bipolar disorders.
Age at onset (AAO) of bipolar disorders (BD) could be influenced both by a repeat length polymorphism (5HTTLPR) in the promoter region of the serotonin transporter gene (SLC6A4) and exposure to childhood trauma. We assessed 308 euthymic patients with BD for the AAO of their first mood episode and childhood trauma. Patients were genotyped for the 5HTTLPR (long/short variant) and the rs25531. Genotypes were classified on functional significance (LL, LS, SS). A sample of 126 Brazilian euthymic patients with BD was used for replication. In the French sample, the correlation between AAO and trauma score was observed only among 'SS' homozygotes (p = 0.002) but not among 'L' allele carriers. A history of at least one trauma decreased the AAO only in 'SS' homozygotes (p = 0.001). These results remained significant after correction using FDR. Regression models suggested an interaction between emotional neglect and 'SS' genotype on the AAO (p = 0.009) and no further interaction with other trauma subtypes. Partial replication was obtained in the Brazilian sample, showing an interaction between emotional abuse and 'LS' genotype on the AAO (p = 0.02). In conclusion, an effect of childhood trauma on AAO of BD was observed only in patients who carry a specific stress responsiveness-related SLC6A4 promoter genotype.


[New recommendations concerning the fluoride content of toddler toothpaste - consequences for systemic application of fluoride].
A group of experts from 4 European countries who gathered at a convention at Basel in November 1998, arrived at the recommendation to increase the fluoride (= F) content of toddler toothpastes from 250 ppm to 500 ppm. It was recommended to make parents brush the children's teeth with a pea-size piece of this toothpaste once a day, starting when the first deciduous teeth were erupting. Routine application of F-tablets would no longer be routinely prescribed, but restricted to individual indications in special high caries risk cases. This recommendation did not consider previous ones and was based exclusively on new scientific, mainly epidemiological evidence. In April 2000 the recommendation was officially issued by the German scientific dental association DGZMK.A careful case-control study resulted in the analysis of the risk to develop mottling of enamel under the influence of fluoridated water (1 ppm F) and fluoride toothpaste (1000 ppm F) when used in early childhood. It was found that excessive use of the fluoride toothpaste doubled the fluorosis risk, whereas when fluoride supplements (tablets, drops) were given the risk was about 20 times higher than without a fluoride supplement. Experiments in Germany and the Netherlands had shown that remineralisation of enamel under influence of 500 ppm F is achieved much more quickly than under application of 250 ppm F. A panel of WHO experts came to the conclusion that there was no evidence for the effectiveness of toothpastes containing less than 500 ppm. Statistics from the Netherlands have shown that the amount of fluoride tablets sold there is barely sufficient for the use by a quarter of all children 0 to 4 years old. In contrast to this low level of acceptance of fluoride tablets, fluoride toothpastes is widely accepted. It is their extensive use which explains the marked improvement of dentitions among the youth in this country during the last 20 years; the influence of topical fluoride gels, varnishes and other preventive measures was much less, and a reduction of sugar consumption (by the way less than 10 % of what it was in 1970) seems to have been the least important factor. The new recommendations based on topical rather than systemic fluoride application are better for preventive, toxicological, psychological and didactic reasons and should be implemented as soon as possible.


The relationship between gamma-glutamyltransferase and adiponectin in nonalcoholic women.
The relationship between adiponectin and gamma-glutamyltransferase (GGT) has yet to be clearly demonstrated especially in women. Among the parameters of the liver function test (LFT), it has become increasingly evident that GGT is associated with metabolic disease. The objective of this study was to characterize the relationship between adiponectin and GGT in nonalcoholic women without liver disease. The subjects in this study were recruited from participants in routine health examinations during February of 2004. Among the total of 115 subjects considered for recruitment, we ultimately included 86 patients without liver disease in the study after performing LFT and abdominal sonography. After a 12-hour overnight fast, levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, GGT, total cholesterol, high-density lipoprotein cholesterol, triglycerides, fasting plasma glucose, fasting insulin, and adiponectin were measured in all subjects. We found a significant negative correlation between adiponectin and GGT (r=-0.35, P<.001) and a significant positive correlation between GGT and homeostasis model assessment (HOMA) (r=0.29, P<.01) after controlling for the confounding influences of age and fat mass. Although GGT is clearly related to adiponectin and HOMA, we determined aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase were not significantly associated with adiponectin and HOMA. The present study suggests that only GGT among the LFTs is related to adiponectin in nonalcoholic women without liver disease.


Autophagy in the renewal, differentiation and homeostasis of immune cells.
Across all branches of the immune system, the process of autophagy is fundamentally important in cellular development, function and homeostasis. Strikingly, this evolutionarily ancient pathway for intracellular recycling has been adapted to enable a high degree of functional complexity and specialization. However, although the requirement for autophagy in normal immune cell function is clear, the mechanisms involved are much less so and encompass control of metabolism, selective degradation of substrates and organelles and participation in cell survival decisions. We review here the crucial functions of autophagy in controlling the differentiation and homeostasis of multiple immune cell types and discuss the potential mechanisms involved.


Cognitive requirements for vestibular and ocular motor processing in healthy adults and patients with unilateral vestibular lesions.
This study investigated the role of cognition in the vestibulo-ocular reflex (VOR) and ocular pursuit using a dual-task paradigm in patients with unilateral peripheral vestibular loss and healthy adults. We hypothesized that cognitive resources are involved in successful processing and integration of vestibular and ocular motor sensory information, and this requirement would be greater in patients with vestibular dysfunction. Sixteen well-compensated patients with surgically confirmed absent unilateral peripheral vestibular function and 16 healthy age- and sex-matched controls underwent seven combinations of vestibular-only, visual-only, and visual-vestibular stimuli while performing three different information processing tasks. Visual-vestibular stimuli included a semicircular canal and an otolith stimulus provided through seated chair rotations; fixation on a laser target and sinusoidal smooth pursuit while still; and fixation on a head-fixed laser target during chair rotations. The information processing tasks were three different auditory reaction time (RT) tasks: (1) simple RT, (2) disjunctive RT, and (3) choice RT. Our results showed increases in RTs in both patients and controls under all vestibular-only stimulation conditions and during ocular pursuit. Patients showed greater increases in RTs during vestibular stimulation and the more complex disjunctive and choice RT tasks. No differences between the groups were found during the visual-only or visual-vestibular interaction conditions. These results reveal interference between vestibulo-ocular processing and a concurrent RT task, suggesting that the VOR and the ocular motor system are dependent upon cognitive resources to some extent, and thus, are not fully automatic systems. We speculate that this interference with cognition occurs as a result of the sensory integration required for resolving inputs from multiple sensory streams. The particularly large decrement in information processing task performance of the patients compared with controls during vestibular stimulation suggests that compensation for unilateral vestibular loss requires continued cognitive resources.


Cerebral blood flow and oxygenation in infants after birth asphyxia. Clinically useful information?
The term 'luxury perfusion' was coined nearly 50 years ago after observation of bright-red blood in the cerebral veins of adults with various brain pathologies. The bright-red blood represents decreased oxygen extraction and hence the perfusion is 'luxurious' compared to oxygen needs. Gradual loss of cellular energy charge during the hours following severe birth asphyxia was observed twenty years later by sequential cranial magnetic resonance spectroscopy. This led to the concept of delayed energy failure that is linked to mitochondrial dysfunction and apoptotic cell death. Abnormally increased perfusion and lack of normal cerebral blood flow regulation are also typically present, but whether the perfusion abnormalities at this secondary stage are detrimental, beneficial, or a mere epiphenomenon remains elusive. In contrast, incomplete reoxygenation of the brain during and following resuscitation is likely to compromise outcome. The clinical value of cerebral oximetry in this context can only be examined in a randomised clinical trial.


Progesterone use to reverse the effects of mifepristone.
To present a series of cases demonstrating successful reversal of mifepristone effects in women who chose to reverse the medical abortion process. Four of 6 women who took mifepristone were able to carry their pregnancies to term after receiving intramuscular progesterone 200 mg. Mifepristone has been available in the US since 2000. By 2008, approximately 25% of abortions prior to 9 weeks were accomplished with mifepristone. Some women who take mifepristone wish to reverse the medical abortion process. Progesterone competes with mifepristone for the progesterone receptor and may reverse the effects of mifepristone. A PubMed literature search from 1996 to May 2012 did not reveal any trials or case studies evaluating the efficacy of progesterone use to reverse the effects of mifepristone. Health care professionals should be aware of the possible use of progesterone to reverse mifepristone in women who have begun the medical abortion process by taking mifepristone and then change their minds.


Responses of bovine early embryos to S-adenosyl methionine supplementation in culture.
There is a growing concern about the potential adverse effects of high dose folic acid (FA) supplementation before and during pregnancy. FA metabolism generates S-adenosyl methionine (SAM) which is an important cofactor of epigenetic programming. We sought to assess the impact of a large dose of SAM on early embryo development. In vitro cultured bovine embryos were treated with SAM from the eight-cell stage to the blastocyst stage. In addition to the phenotype, the genome-wide epigenetic and transcription profiles were analyzed. Treatment significantly improved embryo hatching and caused a shift in sex ratio in favor of males. SAM caused genome-wide hypermethylation mainly in exonic regions and in CpG islands. Although differentially expressed genes were associated with response to nutrients and developmental processes, no correspondence was found with the differentially methylated regions, suggesting that cellular responses to SAM treatment during early embryo development may not require DNA methylation-driven changes. Since bovine embryos were not indifferent to SAM, effects of large-dose FA supplements on early embryonic development in humans cannot be ruled out.


Adipose-derived stem cells and the stromal vascular fraction in polyglycolic acid-collagen nerve conduits promote rat facial nerve regeneration.
Adipose-derived stem cells (ADSCs) and the stromal vascular fraction (SVF) promote nerve regeneration. Biodegradable nerve conduits are used to treat peripheral nerve injuries, but their efficiencies are lower than those of autologous nerve grafts. This study developed biodegradable nerve conduits containing ADSCs and SVF and evaluated their facial nerve regenerating abilities in a rat model with a 7-mm nerve defect. SVF and ADSCs were individually poured into nerve conduits with polyglycolic acid-type I collagen as a scaffold (ADSCs and SVF groups). The conduits were grafted on to the nerve defects. As the control, the defect was bridged with polyglycolic acid-collagen nerve conduits without cells. At 13 weeks, after transplantation, the regenerated nerves were evaluated physiologically and histologically. The compound muscle action potential of the SVF group was significantly higher in amplitude than that of the control group. Electron microscopy showed that the axon diameter of the SVF group was the largest, followed by the ADSC group and control group with significant differences among them. The SVF group had the largest fiber diameter, followed by the ADSC group and control group with significant differences among them. The ADSC group had the highest myelin thickness, followed by the SVF group and control group with significant differences among them. Identical excellent promoting effects on nerve regeneration were observed in both the ADSC and SVF groups. Using SVF in conduits was more practical than using ADSCs because only the enzymatic process was required to prepare SVF, indicating that SVF could be more suitable to induce nerve regeneration.


CD81-induced behavioural changes during chronic cocaine administration: in vivo gene delivery with regulatable lentivirus.
CD81, a tetraspanin transmembrane protein involved in cell adhesion, is up-regulated in the mesolimbic dopaminergic pathway 24 h following acute administration of high doses of cocaine [Brenz-Verca et al., (2001) Mol. Cell. Neurosci., 17, 303-316]. Further evidence consecutive with this observation and based on microarray analysis are presented here. In addition, a regulatable lentivirus was developed bearing the rat CD81 gene under the control of a tetracycline inducible system. This lentivirus vector was stereotaxically injected into the ventral tegmental area (VTA) of two groups of animals, one fed water (expressing CD81) and the other Doxycycline solution (which down-regulates CD81 expression) and locomotor activity after chronic cocaine administration (10 mg/kg daily) was monitored. After 2 weeks, the groups were inverted, animals receiving water were placed on Doxycycline and the second group was placed on water. In all cases highly a significant increase (3.2-fold) in locomotor activity was observed in animals expressing CD81 in the VTA vs. animals placed on Doxycycline. Similar studies where CD81 was delivered into the nucleus accumbens (NAcc) resulted in significantly higher effects (30%), in accordance with microarray data and our previous reports, yielding a 4.2-fold increase in locomotor activity. No change was observed under similar conditions in control animals, which were injected a regulatable lentivirus expressing GFP. These findings suggest that CD81 expression in the mesolimbic dopaminergic pathway contributes to behavioural changes associated with cocaine sensitization. This study provides a powerful approach for evaluating a gene function in vivo in a single animal under various paradigms, even on gene candidates, which display small changes of expression.


Molecular and biochemical characterization of 3-hydroxybenzoate 6-hydroxylase from Polaromonas naphthalenivorans CJ2.
Prior research revealed that Polaromonas naphthalenivorans CJ2 carries and expresses genes encoding the gentisate metabolic pathway for naphthalene. These metabolic genes are split into two clusters, comprising nagRAaGHAbAcAdBFCQEDJI'-orf1-tnpA and nagR2-orf2I''KL (C. O. Jeon, M. Park, H. Ro, W. Park, and E. L. Madsen, Appl. Environ. Microbiol. 72:1086-1095, 2006). BLAST homology searches of sequences in GenBank indicated that the orf2 gene from the small cluster likely encoded a salicylate 5-hydroxylase, presumed to catalyze the conversion of salicylate into gentisate. Here, we report physiological and genetic evidence that orf2 does not encode salicylate 5-hydroxylase. Instead, we have found that orf2 encodes 3-hydroxybenzoate 6-hydroxylase, the enzyme which catalyzes the NADH-dependent conversion of 3-hydroxybenzoate into gentisate. Accordingly, we have renamed orf2 nagX. After expression in Escherichia coli, the NagX enzyme had an approximate molecular mass of 43 kDa, as estimated by gel filtration, and was probably a monomeric protein. The enzyme was able to convert 3-hydroxybenzoate into gentisate without salicylate 5-hydroxylase activity. Like other 3-hydroxybenzoate 6-hydroxylases, NagX utilized both NADH and NADPH as electron donors and exhibited a yellowish color, indicative of a bound flavin adenine dinucleotide. An engineered mutant of P. naphthalenivorans CJ2 defective in nagX failed to grow on 3-hydroxybenzoate but grew normally on naphthalene. These results indicate that the previously described small catabolic cluster in strain CJ2 may be multifunctional and is essential for the degradation of 3-hydroxybenzoate. Because nagX and an adjacent MarR-type regulatory gene are both closely related to homologues in Azoarcus species, this study raises questions about horizontal gene transfer events that contribute to operon evolution.


Novel class of Bi(iii) hydroxamato complexes: synthesis, urease inhibitory activity and activity against H. pylori.
Reaction of Bi(NO3)3 with benzohydroxamic acid (Bha) and salicylhydroxamic acid (Sha) gives the novel Bi(iii) complexes [Bi2(Bha-1H)2(μ-Bha-1H)2(η(2)-NO3)2] () and [Bi6(CH3OH)2(η(1)-NO3)2(η(2)-NO3)(OH2)2(Sha-1H)12](NO3)2 (). X-ray crystal structure of reveals two hydroxamato coordination modes; bidentate bridging (O, O') and bidentate non-bridging (O, O') and of reveals one coordination mode; bidentate bridging (O, O'). , specifically designed to and demonstrated to inhibit the activity of urease, exhibits excellent antibacterial activity against three strains of Helicobacter pylori with MIC ≥ 16 μg mL(-1).


Polyomavirus-plasmid recombinants capable of replicating have an enhanced transforming potential.
The frequency of transformation of rodent fibroblasts by polyomavirus is enhanced by a viral gene product, large T-antigen. However, this effect of large T-antigen cannot be demonstrated with pBR322-cloned viral DNA. Recently, it was discovered that pBR322 contains cis-acting sequences inhibitory to DNA replication in mammalian cells. Because polyomavirus large T-antigen is required for viral DNA replication, we examined the possibility that our inability to demonstrate a requirement for large T-antigen in transformation with pBR322-cloned viral DNA was due to the failure of the chimeric DNA to replicate in the transfected cells. To this end we constructed polyomavirus recombinant molecules with a plasmid (pML-2) that lacks these "poison" sequences and measured their capacity to transform cells. Here we report that recombinant plasmids capable of replicating in the transfected cells transform these cells at frequencies approximately sixfold greater than their replication-defective counterparts.


Effect of 18F-Fluciclovine Positron Emission Tomography on the Management of Patients With Recurrence of Prostate Cancer: Results From the FALCON Trial.
Early and accurate localization of lesions in patients with biochemical recurrence (BCR) of prostate cancer may guide salvage therapy decisions. The present study, 18F-Fluciclovine PET/CT in biochemicAL reCurrence Of Prostate caNcer (FALCON; NCT02578940), aimed to evaluate the effect of 18F-fluciclovine on management of men with BCR of prostate cancer. Men with a first episode of BCR after curative-intent primary therapy were enrolled at 6 UK sites. Patients underwent 18F-fluciclovine positron emission tomography/computed tomography (PET/CT) according to standardized procedures. Clinicians documented management plans before and after scanning, recording changes to treatment modality as major and changes within a modality as other. The primary outcome measure was record of a revised management plan postscan. Secondary endpoints were evaluation of optimal prostate specific antigen (PSA) threshold for detection, salvage treatment outcome assessment based on 18F-fluciclovine-involvement, and safety. 18F-Fluciclovine was well tolerated in the 104 scanned patients (median PSA = 0.79 ng/mL). Lesions were detected in 58 out of 104 (56%) patients. Detection was broadly proportional to PSA level; ≤1 ng/mL, 1 out of 3 of scans were positive, and 93% scans were positive at PSA >2.0 ng/mL. Sixty-six (64%) patients had a postscan management change (80% after a positive result). Major changes (43 out of 66; 65%) were salvage or systemic therapy to watchful waiting (16 out of 66; 24%); salvage therapy to systemic therapy (16 out of 66; 24%); and alternative changes to treatment modality (11 out of 66, 17%). The remaining 23 out of 66 (35%) management changes were modifications of the prescan plan: most (22 out of 66; 33%) were adjustments to planned brachytherapy/radiation therapy to include a 18F-fluciclovine-guided boost. Where 18F-fluciclovine guided salvage therapy, the PSA response rate was higher than when 18F-fluciclovine was not involved (15 out of 17 [88%] vs 28 out of 39 [72%]). 18F-Fluciclovine PET/CT located recurrence in the majority of men with BCR, frequently resulting in major management plan changes. Incorporating 18F-fluciclovine PET/CT into treatment planning may optimize targeting of recurrence sites and avoid futile salvage therapy.


Design and functional characterization of a novel, arrestin-biased designer G protein-coupled receptor.
Mutational modification of distinct muscarinic receptor subtypes has yielded novel designer G protein-coupled receptors (GPCRs) that are unable to bind acetylcholine (ACh), the endogenous muscarinic receptor ligand, but can be efficiently activated by clozapine-N-oxide (CNO), an otherwise pharmacologically inert compound. These CNO-sensitive designer GPCRs [alternative name: designer receptors exclusively activated by designer drug (DREADDs)] have emerged as powerful new tools to dissect the in vivo roles of distinct G protein signaling pathways in specific cell types or tissues. As is the case with other GPCRs, CNO-activated DREADDs not only couple to heterotrimeric G proteins but can also recruit proteins of the arrestin family (arrestin-2 and -3). Accumulating evidence suggests that arrestins can act as scaffolding proteins to promote signaling through G protein-independent signaling pathways. To explore the physiological relevance of these arrestin-dependent signaling pathways, the availability of an arrestin-biased DREADD would be highly desirable. In this study, we describe the development of an M₃ muscarinic receptor-based DREADD [Rq(R165L)] that is no longer able to couple to G proteins but can recruit arrestins and promote extracellular signal-regulated kinase-1/2 phosphorylation in an arrestin- and CNO-dependent fashion. Moreover, CNO treatment of mouse insulinoma (MIN6) cells expressing the Rq(R165L) construct resulted in a robust, arrestin-dependent stimulation of insulin release, directly implicating arrestin signaling in the regulation of insulin secretion. This newly developed arrestin-biased DREADD represents an excellent novel tool to explore the physiological relevance of arrestin signaling pathways in distinct tissues and cell types.


Neurophysiological effects of mobile phone electromagnetic fields on humans: a comprehensive review.
In recent years a growing number of people have begun to use mobile phone technology. This phenomenon has raised questions and doubts about possible effects on users' brains. This literature review focuses on the human electrophysiological and neuro-metabolic effects of mobile phone (MP)-related electromagnetic fields (EMFs) published in the last 10 years. To this end, all relevant papers have been reported and, subsequently, a literature selection has been carried out by taking several criteria into account, such as: blind techniques, randomization or counter-balancing of conditions and subjects, detail of exposure characteristics and the statistical analyses used. As a result, only the studies meeting the selection criteria have been described, evaluated and discussed further. The main goal of this review is to provide a clear scenario of the most reliable experiments carried out over the last decade and to offer a critical point of view in their evaluation. It is concluded that MP-EMFs may influence normal physiology through changes in cortical excitability and that in future research particular care should be dedicated to both methodological and statistical control, the most relevant criteria in this research field.


The combination of ursodeoxycholic acid and methotrexate for patients with primary biliary cirrhosis: the results of a pilot study.
Ursodeoxycholic acid (UDCA) and methotrexate (MTX) have both been proposed as treatments for patients with primary biliary cirrhosis (PBC). It has been suggested that a combination of the two drugs may offer advantages over either used separately. In this pilot study, we sought to evaluate the safety and efficacy of this combination for patients with PBC. Thirty-two patients with antimitochondrial antibody positive PBC were prospectively entered into a pilot study and received UDCA, 13 to 15 mg/kg/d, in conjunction with MTX, 0.25 mg/kg/wk, for a period of 2 years. The results of this treatment were compared with those obtained from 180 patients with PBC studied in a placebo-controlled trial of UDCA alone conducted during the same period. Patients in the pilot study and randomized study were comparable with regard to age, gender, and liver biochemistries. The UDCA/MTX-treated patients were of earlier histologic stage and had a lower mean Mayo risk score. During this period, seven patients in the UDCA/MTX group were withdrawn, four for pulmonary toxicity (two who required hospitalization), and one each with mouth ulcer, extreme fatigue, and hair loss. The use of UDCA/MTX was not associated with improvement in symptoms. In the patients receiving UDCA/MTX, biochemical changes were comparable to those of patients receiving UDCA alone but superior to those in the placebo group (P < .05). Histological changes were comparable in all groups at 2 years. Cessation of MTX while UDCA was continued led to no deterioration in liver biochemistries.(ABSTRACT TRUNCATED AT 250 WORDS)


The Hermann grid illusion revisited.
The Hermann grid illusion consists of smudges perceived at the intersections of a white grid presented on a black background. In 1960 the effect was first explained by a theory advanced by Baumgartner suggesting the illusory effect is due to differences in the discharge characteristics of retinal ganglion cells when their receptive fields fall along the intersections versus when they fall along non-intersecting regions of the grid. Since then, others have claimed that this theory might not be adequate, suggesting that a model based on cortical mechanisms is necessary [Lingelbach et al, 1985 Perception 14(1) A7; Spillmann, 1994 Perception 23 691 708; Geier et al, 2004 Perception 33 Supplement, 53; Westheimer, 2004 Vision Research 44 2457 2465]. We present in this paper the following evidence to show that the retinal ganglion cell theory is untenable: (i) varying the makeup of the grid in a manner that does not materially affect the putative differential responses of the ganglion cells can reduce or eliminate the illusory effect; (ii) varying the grid such as to affect the putative differential responses of the ganglion cells does not eliminate the illusory effect; and (iii) the actual spatial layout of the retinal ganglion cell receptive fields is other than that assumed by the theory. To account for the Hermann grid illusion we propose an alternative theory according to which the illusory effect is brought about by the manner in which S1 type simple cells (as defined by Schiller et al, 1976 Journal of Neurophysiology 39 1320-1333) in primary visual cortex respond to the grid. This theory adequately handles many of the facts delineated in this paper.


Mechanical regulation of cardiac myofibrillar structure.
The excitation-contraction coupling cycle (ECC) consists of a complex cascade of electrochemical and mechanical events; however, the relative contributions of these different processes in the regulation of cardiac myofibrillar structure are not well understood. There is extensive evidence to suggest that the mechanical aspects of the ECC play a crucial role in controlling the availability of contractile proteins for myofibrillar assembly. To examine if these physical forces might also serve to stabilize the structure of preexisting myofibrils, beating and nonbeating cultures of neonatal cardiac myocytes (NCM) were subjected to a 5% static stretch. Contractile arrest was achieved by treating NCM with 12 microM nifedipine, which resulted in immediate and sustained contractile arrest and initiated the evolution of marked myofibrillar abnormalities within 24 hours. As judged by scanning confocal and transmission electron microscopic examination, an external load appears to partially stabilize myofibrillar structure in nonbeating NCM. These results suggest that the maintenance of myofibrillar structure may be highly dependent upon the mechanical aspects of ECC.


Narrowband photodetection in the near-infrared with a plasmon-induced hot electron device.
In gratings, incident light can couple strongly to plasmons propagating through periodically spaced slits in a metal film, resulting in a strong, resonant absorption whose frequency is determined by the nanostructure periodicity. When a grating is patterned on a silicon substrate, the absorption response can be combined with plasmon-induced hot electron photocurrent generation. This yields a photodetector with a strongly resonant, narrowband photocurrent response in the infrared, limited at low frequencies by the Schottky barrier, not the bandgap of silicon. Here we report a grating-based hot electron device with significantly larger photocurrent responsivity than previously reported antenna-based geometries. The grating geometry also enables more than three times narrower spectral response than observed for nanoantenna-based devices. This approach opens up the possibility of plasmonic sensors with direct electrical readout, such as an on-chip surface plasmon resonance detector driven at a single wavelength.


TeleWound Practice Within the Veterans Health Administration: Protocol for a Mixed Methods Program Evaluation.
Chronic wounds, such as pressure injuries and diabetic foot ulcers, are a significant predictor of mortality. Veterans who reside in rural areas often have difficulty accessing care for their wounds. TeleWound Practice (TWP), a coordinated effort to incorporate telehealth into the provision of specialty care for patients with skin wounds, has the potential to increase access to wound care by allowing veterans to receive this care at nearby outpatient clinics or in their homes. The Veterans Health Administration (VA) is championing the rollout of the TWP, starting with regional implementation. This paper aims to describe the protocol for a mixed-methods program evaluation to assess the implementation and outcomes of TWP in VA. We are conducting a mixed-methods evaluation of 4 VA medical centers and their community-based outpatient clinics that are participating in the initial implementation of the TWP. Data will be collected from veterans, VA health care team members, and other key stakeholders (eg, clinical leadership). We will use qualitative methods (ie, semistructured interviews), site visits, and quantitative methods (ie, surveys, national VA administrative databases) to assess the process and reach of TWP implementation and its impact on veterans' clinical outcomes and travel burdens and costs. This program evaluation was funded in October 2019 as a Partnered Evaluation Initiative by the US Department of Veterans Affairs, Diffusion of Excellence Office, and Office of Research and Development, Health Services Research and Development Service, Quality Enhancement Research Initiative Program (PEC 19-310). Evaluation of the TWP will identify barriers and solutions to TeleWound implementation in a small number of sites that can be used to inform successful rollout of the TWP nationally. Our evaluation work will inform future efforts to scale up the TWP across VA and optimize reach of the program to veterans across the nation. DERR1-10.2196/20139.


Functional genomic profiling of Aspergillus fumigatus biofilm reveals enhanced production of the mycotoxin gliotoxin.
The opportunistic pathogenic mold Aspergillus fumigatus is an increasing cause of morbidity and mortality in immunocompromised and in part immunocompetent patients. A. fumigatus can grow in multicellular communities by the formation of a hyphal network encased in an extracellular matrix. Here, we describe the proteome and transcriptome of planktonic- and biofilm-grown A. fumigatus mycelium after 24 and 48 h. A biofilm- and time-dependent regulation of many proteins and genes of the primary metabolism indicates a developmental stage of the young biofilm at 24 h, which demands energy. At a matured biofilm phase, metabolic activity seems to be reduced. However, genes, which code for hydrophobins, and proteins involved in the biosynthesis of secondary metabolites were significantly upregulated. In particular, proteins of the gliotoxin secondary metabolite gene cluster were induced in biofilm cultures. This was confirmed by real-time PCR and by detection of this immunologically active mycotoxin in culture supernatants using HPLC analysis. The enhanced production of gliotoxin by in vitro formed biofilms reported here may also play a significant role under in vivo conditions. It may confer A. fumigatus protection from the host immune system and also enable its survival and persistence in chronic lung infections such as aspergilloma.


The Risks of Revolution: Ethical Dilemmas in 3D Printing from a US Perspective.
Additive manufacturing has spread widely over the past decade, especially with the availability of home 3D printers. In the future, many items may be manufactured at home, which raises two ethical issues. First, there are questions of safety. Our current safety regulations depend on centralized manufacturing assumptions; they will be difficult to enforce on this new model of manufacturing. Using current US law as an example, I argue that consumers are not capable of fully assessing all relevant risks and thus continue to require protection; any regulation will likely apply to plans, however, not physical objects. Second, there are intellectual property issues. In combination with a 3D scanner, it is now possible to scan items and print copies; many items are not protected from this by current intellectual property laws. I argue that these laws are ethically sufficient. Patent exists to protect what is innovative; the rest is properly not protected. Intellectual property rests on the notion of creativity, but what counts as creative changes with the rise of new technologies.


Erratic flu vaccination emerges from short-sighted behavior in contact networks.
The effectiveness of seasonal influenza vaccination programs depends on individual-level compliance. Perceptions about risks associated with infection and vaccination can strongly influence vaccination decisions and thus the ultimate course of an epidemic. Here we investigate the interplay between contact patterns, influenza-related behavior, and disease dynamics by incorporating game theory into network models. When individuals make decisions based on past epidemics, we find that individuals with many contacts vaccinate, whereas individuals with few contacts do not. However, the threshold number of contacts above which to vaccinate is highly dependent on the overall network structure of the population and has the potential to oscillate more wildly than has been observed empirically. When we increase the number of prior seasons that individuals recall when making vaccination decisions, behavior and thus disease dynamics become less variable. For some networks, we also find that higher flu transmission rates may, counterintuitively, lead to lower (vaccine-mediated) disease prevalence. Our work demonstrates that rich and complex dynamics can result from the interaction between infectious diseases, human contact patterns, and behavior.


Transforming growth factor beta and interleukin-1: a paradigm for opposing regulation of haemopoiesis.
The polypeptide cytokines, IL-1 and TGF-beta affect nearly every tissue and cell type in the body. IL-1 is the prototype of the proinflammatory molecule while TGF-beta is essentially anti-inflammatory. IL-1 is part of the cascade of cytokines that are produced during microbial invasion or bodily injury and enhance a variety of host responses, particularly in the immunological and haemopoietic systems, while TGF-beta acts as an inhibitor of these responses. At several levels, IL-1 and TGF-beta act in opposition to one another. IL-1 stimulates the expression of many genes in lymphoid and marrow stromal cells that stimulate haemopoietic cell growth and differentiation, while TGF-beta inhibits these IL-1 mediated effects. Also, TGF-beta stimulates secretion of the IL-1Ra. In addition, IL-1 induces the cell surface expression of cytokine receptors on lymphoid and haemopoietic cells, while TGF-beta dramatically inhibits the cell surface expression of these receptors, including the IL-1 receptor. Finally, IL-1 augments lymphoid and haemopoietic cell growth and TGF-beta potently inhibits this proliferation. The interactions of these cytokines serve to illustrate that the net balance of stimulatory and inhibitory signals determines the fate of a given cell which may be responsible for regulating homeostatic cell growth (Figure 1). Thus, the regulation of cytokine production and/or antagonism of their effects continues to be a therapeutic goal in the treatment of many diseases.


(-)-Englerin A is a potent and selective activator of TRPC4 and TRPC5 calcium channels.
Current therapies for common types of cancer such as renal cell cancer are often ineffective and unspecific, and novel pharmacological targets and approaches are in high demand. Here we show the unexpected possibility for the rapid and selective killing of renal cancer cells through activation of calcium-permeable nonselective transient receptor potential canonical (TRPC) calcium channels by the sesquiterpene (-)-englerin A. This compound was found to be a highly efficient, fast-acting, potent, selective, and direct stimulator of TRPC4 and TRPC5 channels. TRPC4/5 activation through a high-affinity extracellular (-)-englerin A binding site may open up novel opportunities for drug discovery aimed at renal cancer.


Ultrastructural examination of one-step self-etch adhesive bonded primary sound and caries-affected dentin.
To examine the ultrastructure and silver nanoleakage of the resin-dentin interfaces in sound and caries-affected primary tooth dentin bonded with a 4-META one-step self-etch adhesive. Each of five sound and carious primary molars was bonded with Hybrid Bond. Resin-dentin interfaces were observed with TEM micrographs obtained from silver-impregnated, unstained and undemineralized sections of bonded sound and caries-affected primary dentin, and stained and demineralized sections of bonded sound and caries affected primary dentin without silver impregnation. For sound dentin, silver nanoleakage was observed extensively in the patent dentin tubules, within the dentin beneath the hybrid layer, within the hybrid layer in some specimens, and as water trees that partially protruded into the overlying adhesive layer. The hybrid layer was about 1 microm thick. Smear plugs in the dentin tubules and smear on the ground dentin protruded in the hybrid layer. Remnants of demineralized smear were observed overlying the adhesive layer. For caries-affected dentin, the hybrid layer was obscure. Dentin tubules were occluded with mineral deposits. There were no water trees or nanoleakage in the adhesive layer or hybrid layer. However, smear remnants were observed in the adhesive layer and large silver deposits were observed in the highly porous underlying caries-affected dentin.


Lifestyle health promotion interventions for the nursing workforce: a systematic review.
Study aims were to identify the efficacy of lifestyle health promotion interventions intended to improve behavioural health risk factors and/or behavioural or clinical outcomes of working-age nurses. Nurses constitute around half the health workforce but global shortages and an ageing profile challenge future supply. The occupational hazards and stresses of nursing are well known. Health promotion, possibly workplace-based, presents opportunities to safeguard the health of nurses. This was a systematic review undertaken in line with guidance for reviews in health care. Seven electronic databases were searched from 2000-2011 and references of relevant papers. Two reviewers independently reviewed and critiqued retrieved papers and extracted data. Methodological features were described using the CONSORT checklists; risk of bias was assessed using the Cochrane Handbook classification. With design inclusion criteria relaxed to include an uncontrolled trial, only three intervention studies were retrieved, from the United States, Canada and Taiwan. All had limitations and high risk of bias, but benefits were reported. Outcomes included fewer cigarettes smoked during the intervention period, down from mean (SD) 20 (8)-12 (9) per day (p < 0·001); significantly reduced fat mass (0·68 vs. 0·07 kg; p = 0·028); and significant gains across a battery of fitness assessments. The paucity of work focused on nurses' health behaviours was the important finding. The workplace is a potentially fruitful location for health promotion intervention but nurses have seldom been recognised as a target participant group. Given the international priority ascribed to nursing workforce retention, this is a missed opportunity for occupational health planning. Potential benefits to nurses' welfare and well-being may accrue from well-designed intervention studies. Nurse leaders have a key role in driving recognition, spearheading commitment and development of targeted, whole-organisation programmes to promote health profile improvement for the nursing workforce.


Maternal stress and the white coat syndrome: a case study.
Coping with the birth of a preterm infant is a stressful event for mothers for many reasons, including parental role alterations, perception of the infant's illness severity, and uncertainty about the infant's prognosis. Stressors evoke many physiological responses. Fingertip temperature, an electrophysiologic indicator, is a good measure of stress. This case study describes the experience of one mother's negative response to a surgeon's visit to her baby's bedside. Researchers refer to the "white coat" syndrome, in which personnel can significantly affect autonomic function. Increasing our understanding of maternal stress could help ameliorate some of the deleterious effects persistently elevated levels of stress can produce. Further research might focus on the effects of communication with other familiar health care professionals, as well as the influence of Kangaroo Care (skin-to-skin, chest-to-chest contact between caregiver and infant) on maternal stress.


Contribution of Adipose Tissue Inflammation to the Development of Type 2 Diabetes Mellitus.
The objective of this comprehensive review is to summarize and discuss the available evidence of how adipose tissue inflammation affects insulin sensitivity and glucose tolerance. Low-grade, chronic adipose tissue inflammation is characterized by infiltration of macrophages and other immune cell populations into adipose tissue, and a shift toward more proinflammatory subtypes of leukocytes. The infiltration of proinflammatory cells in adipose tissue is associated with an increased production of key chemokines such as C-C motif chemokine ligand 2, proinflammatory cytokines including tumor necrosis factor α and interleukins 1β and 6 as well as reduced expression of the key insulin-sensitizing adipokine, adiponectin. In both rodent models and humans, adipose tissue inflammation is consistently associated with excess fat mass and insulin resistance. In humans, associations with insulin resistance are stronger and more consistent for inflammation in visceral as opposed to subcutaneous fat. Further, genetic alterations in mouse models of obesity that reduce adipose tissue inflammation are-almost without exception-associated with improved insulin sensitivity. However, a dissociation between adipose tissue inflammation and insulin resistance can be observed in very few rodent models of obesity as well as in humans following bariatric surgery- or low-calorie-diet-induced weight loss, illustrating that the etiology of insulin resistance is multifactorial. Taken together, adipose tissue inflammation is a key factor in the development of insulin resistance and type 2 diabetes in obesity, along with other factors that likely include inflammation and fat accumulation in other metabolically active tissues. © 2019 American Physiological Society. Compr Physiol 9:1-58, 2019.


Influence of the composition of natural organic matter on Pb bioavailability to microalgae.
The current work examines the effects of model allochtonous (humic substances) and autochtonous (microbial polysaccharides) natural organic matter (NOM) on Pb speciation and bioaccumulation. The results demonstrated that polysaccharides, in particular alginic acid, had complexing properties and effects on Pb bioaccumulation by the green alga Chlorella kesslerii that were similar to equivalent complexing capacity of humic substances. Pb uptake decreased in the presence of humic, alginic, and polygalacturonic acids with respect to noncomplexed Pb, but accumulated Pb was higher than predicted from measured Pb2+ concentrations or from previous results obtained in the presence of simple synthetic ligands. An improved fit between experimental observations and Pb speciation was obtained by taking into account the formation of a ternary complex at the algal surface. The contribution of the ternary complexes to Pb bioaccumulation was dependent on the relative binding constants of the Pb to the NOM and to the binding sites on the biological surface. In the presence of the humic acid, a decreased surface charge and increased membrane permeability were considered to be of secondary importance to explain the observation of increased Pb uptake with respect to that predicted on the basis of [Pb2+]. The environmental implications of the results are discussed with respect to the development of site-specific water quality criteria.


Antiarrhythmic effect of lithium in rats after myocardial infarction by activation of Nrf2/HO-1 signaling.
Glycogen synthase kinase-3 (GSK-3) signaling has been shown to play a role in the regulation of nuclear factor erythroid-2-related factor 2 (Nrf2), a master regulator of antioxidant genes, including heme oxygenase-1 (HO-1). We assessed whether lithium, a GSK-3 inhibitor, attenuates cardiac sympathetic reinnervation after myocardial infarction, a status of high reactive oxygen species (ROS), by attenuating nerve growth factor (NGF) expression and whether Nrf2/HO-1 signaling is involved in the protection. Twenty-four hours after ligation of the left anterior descending artery, male Wistar rats were treated for 4 weeks. The postinfarction period was associated with increased oxidative-nitrosative stress, as measured by myocardial superoxide, nitrotyrosine, and dihydroethidium fluorescent staining. In concert, myocardial norepinephrine levels and immunohistochemical analysis of sympathetic nerve revealed a significant increase in innervation in vehicle-treated rats compared with sham-operated rats. Arrhythmic scores during programmed stimulation in the vehicle-treated rats were significantly higher than those in sham. This was paralleled by a significant upregulation of NGF protein and mRNA in the vehicle-treated rats, which was reduced after administration of LiCl. LiCl stimulated the nuclear translocation of Nrf2 and the transactivation of the Nrf2 target gene HO-1. Inhibition of phosphoinositide 3-kinase by wortmannin reduced the increase in Nrf2 nucleus translocation and HO-1 expression compared with lithium alone. In addition, the lithium-attenuated NGF levels were reversed in the presence of the Nrf2 inhibitor trigonelline, HO-1 inhibitor SnPP, and peroxynitrite generator SIN-1, indicating the role of Nrf2/HO-1/ROS. In conclusion, lithium protects against ventricular arrhythmias by attenuating NGF-induced sympathetic innervation via antioxidant activation of the Nrf2/HO-1 axis.


Tumor lymphangiogenesis: a novel prognostic indicator for cutaneous melanoma metastasis and survival.
Malignant melanomas of the skin are distinguished by their propensity for early metastatic spread via lymphatic vessels to regional lymph nodes, and lymph node metastasis is a major determinant for the staging and clinical management of melanoma. However, the importance of tumor-induced lymphangiogenesis for lymphatic melanoma spread has remained unclear. We investigated whether tumor lymphangiogenesis occurs in human malignant melanomas of the skin and whether the extent of tumor lymphangiogenesis may be related to the risk for lymph node metastasis and to patient survival, using double immunostains for the novel lymphatic endothelial marker LYVE-1 and for the panvascular marker CD31. Tumor samples were obtained from clinically and histologically closely matched cases of primary melanomas with early lymph node metastasis (n = 18) and from nonmetastatic melanomas (n = 19). Hot spots of proliferating intratumoral and peritumoral lymphatic vessels were detected in a large number of melanomas. The incidence of intratumoral lymphatics was significantly higher in metastatic melanomas and correlated with poor disease-free survival. Metastatic melanomas had significantly more and larger tumor-associated lymphatic vessels, and a relative lymphatic vessel area of >1.5% was significantly associated with poor disease-free and overall survival. In contrast, no differences in the density of tumor-associated blood vessels were found. Vascular endothelial growth factor and vascular endothelial growth factor-C expression was equally detected in a minority of cases in both groups. Our results reveal tumor lymphangiogenesis as a novel prognostic indicator for the risk of lymph node metastasis in cutaneous melanoma.


Development of a Rapid and Precise Reversed-phase High-performance Liquid Chromatography Method for Analysis of Docetaxel in Rat Plasma: Application in Single-dose Pharmacokinetic Studies of Folate-targeted Micelles Containing Docetaxel.
A simple and sensitive reversed-phase high-performance liquid chromatography (HPLC) method based on liquid-liquid extraction was established and validated for determination of docetaxel (DTX) in plasma of rat. Samples were spiked with paclitaxel as the internal standard and the chromatographic separation was carried out using C18 HPLC column. The mobile phase consisted of a mixture of acetonitrile/water with the ratio of 60/40 v/v. The ultraviolet detector was operated at 230 nm, and the flow rate of mobile phase was 1 ml/min. The method was validated for linearity, precision, accuracy, recovery, and limit of quantification (LOQ). Then the method was applied to quantify DTX in the rat plasma after intravenous (IV) administration of the self-assembled micelles of folate-targeted Synpronic F127/cholesterol (FA-PF127-Chol) loaded with DTX and Taxotere® as the reference marketed solution of DTX. The blood samples were taken from the ophthalmic vein at predetermined time intervals after treatment. Calibration curve was linear between the concentration ranges of 0.1-7.5 μg/ml with the relative standard deviation % and evaluating error % ranged from 2.263 to 15.53 and -12.75 to 12.7 for intra- and inter-day validity, respectively. The mean recovery of the drug after plasma extraction was 95.67 ± 0.99% for the concentration of 1 μg/ml. The LOQ and the limit of detection for DTX in serum were 100 ng/ml and 30 ng/ml, respectively. The results indicated that the developed method could be adopted for pharmacokinetic studies of DTX-loaded FA-PF127-Chol micelles and Taxotere® in rat.


Will the global HIV response fail gay and bisexual men and other men who have sex with men?
Gay and bisexual men and other men who have sex with men are among the small number of groups for whom HIV remains uncontrolled worldwide. Although there have been recent and notable decreases in HIV incidence across several countries, prevalence and incidence is consistently higher or rising among men who have sex with men when compared with other groups. In 2014, MSMGF (the Global Forum on MSM & HIV) conducted its third biennial Global Men's Health and Rights Study, an international, multilingual, web-based cross-sectional survey of men who have sex with men recruited through online convenience sampling. We tested hypothesized correlates (selected a priori) of successfully achieving each step along the HIV prevention and treatment continuum by fitting separate generalized estimating equation models adjusted for clustering by country in multivariate analyses. All models controlled for ability to meet basic financial needs, age, healthcare coverage, having a regular provider, region and country-level income. Higher provider discrimination and sexual stigma were associated with lower odds of perceived access to services, service utilization and virologic suppression. Conversely, accessing services from community-based organizations focused on lesbian, gay, bisexual and transgender people; greater engagement in gay community; and comfort with healthcare providers were associated with higher odds of achieving steps along the prevention and treatment continuum. To meet accelerated global HIV targets, global leaders must adopt a differentiated and bolder response, in keeping with current epidemiologic trends and community-based research. The HIV-related needs of gay and bisexual men and other men who have sex with men must be addressed openly, quickly and with sufficient resources to support evidence-based, community-led and human rights-affirming interventions at scale.


Distinctive personality profiles of fibromyalgia and chronic fatigue syndrome patients.
The current study is an innovative exploratory investigation, aiming at identifying differences in personality profiles within Fibromyalgia Syndrome (FMS) and Chronic Fatigue Syndrome (CFS) patients. In total, 344 participants (309 female, 35 male) reported suffering from FMS and/or CFS and consented to participate in the study. Participants were recruited at an Israeli FM/CFS patient meeting held in May 2013, and through an announcement posted on several social networks. Participants were asked to complete a research questionnaire, which included FMS criteria and severity scales, and measures of personality, emotional functioning, positivity, social support and subjective assessment of general health. In total, 204 participants completed the research questionnaire (40.7% attrition rate). A cluster analysis produced two distinct clusters, which differed significantly on psychological variables, but did not differ on demographic variables or illness severity. As compared to cluster number 2 (N = 107), participants classified into cluster number 1 (N = 97) showed a less adaptive pattern, with higher levels of Harm Avoidance and Alexithymia; higher prevalence of Type D personality; and lower levels of Persistence (PS), Reward dependence (RD), Cooperation, Self-directedness (SD), social support and positivity. The significant pattern of results indicates at least two distinct personality profiles of FM and CFS patients. Findings from this research may help improve the evaluation and treatment of FM and CFS patients, based on each patient's unique needs, psychological resources and weaknesses, as proposed by the current trend of personalized medicine.


Shiga toxin-producing Escherichia coli strains from bovines: association of adhesion with carriage of eae and other genes.
Out of 174 bovine Shiga toxin-producing Escherichia coli (STEC) strains isolated from diarrheic calves in Germany and Belgium, 122 strains (70.1%) were selected because of their reactivity with the eae (E. coli attaching and effacing gene) probe ECW1-ECW2. One hundred seven of these eae-positive strains (87.7%) harbored stx1 genes, 13 strains (10.7%) had stx2 genes, and 2 strains (1.6%) had both stx genes. The strains displayed 17 different O types, the majority (97 strains) [79.5%]) belonging to O5 (5 strains), O26 (21 strains), O111 (13 strains) O118 (36 strains), O145 (9 strains), and O157 (13 strains). In the HEp-2 cell adhesion assay, 99 strains (81.1%) showed a localized adhesion, and 80 strains (65.6%) stimulated actin accumulation, as determined in the fluorescence actin staining test. None of the strains harbored genes coding for bundle-forming pili (bfpA), clearly differentiating them from enteropathogenic. E. cole. espB gene sequences were only detectable in 23 (18.9%) of the eae-positive bovine STEC strains. Three different PCRs were established, differentiating between eae sequences of enteropathogenic E. coli strain E2348/69 (O127:H6) and STEC strain EDL933 (O157: H7). Primers matching in the more heterologous downstream eae sequences gave amplicons in only 8 of the 17 O types (O84:H-, O103:H2, O111:H-, O111:H2, O119:H25, O128:H-, O145:H28, and O157:H-). Only 15 STEC strains, belonging to serotypes O111H:-, O111H:2, O145:H28, and O157:H-, gave amplicons in all three eae-specific PCRs. These data demonstrate that bovine STEC strains are a heterogeneous group of pathogenic bacteria, a lot of which share virulence markers with STEC strains causing infections in humans. However, in contrast to human STEC strains, bovine eae-positive STEC strains are mainly restricted to the stx1 genotype. The observation that espB sequences are not highly conserved might have consequences for the serological recognition of the ESPB protein in patients. Like in human STEC strains, eae-related sequences are closely associated with certain E. coli O groups; however, they are not serotype specific.


Slowing the Kinetics of Alumina Sol-Gel Chemistry for Controlled Catalyst Overcoating and Improved Catalyst Stability and Selectivity.
Catalyst overcoating is an emerging approach to engineer surface functionalities on supported metal catalyst and improve catalyst selectivity and durability. Alumina deposition on high surface area material by sol-gel chemistry is traditionally difficult to control due to the fast hydrolysis kinetics of aluminum-alkoxide precursors. Here, sol-gel chemistry methods are adapted to slow down these kinetics and deposit nanometer-scale alumina overcoats. The alumina overcoats are comparable in conformality and thickness control to overcoats prepared by atomic layer deposition even on high surface area substrates. The strategy relies on regulating the hydrolysis/condensation kinetics of Al(s BuO)3 by either adding a chelating agent or using nonhydrolytic sol-gel chemistry. These two approaches produce overcoats with similar chemical properties but distinct physical textures. With chelation chemistry, a mild method compatible with supported base metal catalysts, a conformal yet porous overcoat leads to a highly sintering-resistant Cu catalyst for liquid-phase furfural hydrogenation. With the nonhydrolytic sol-gel route, a denser Al2 O3 overcoat can be deposited to create a high density of Lewis acid-metal interface sites over Pt on mesoporous silica. The resulting material has a substantially increased hydrodeoxygenation activity for the conversion of lignin-derived 4-propylguaiacol into propylcyclohexane with up to 87% selectivity.


Learning nonsparse kernels by self-organizing maps for structured data.
The development of neural network (NN) models able to encode structured input, and the more recent definition of kernels for structures, makes it possible to directly apply machine learning approaches to generic structured data. However, the effectiveness of a kernel can depend on its sparsity with respect to a specific data set. In fact, the accuracy of a kernel method typically reduces as the kernel sparsity increases. The sparsity problem is particularly common in structured domains involving discrete variables which may take on many different values. In this paper, we explore this issue on two well-known kernels for trees, and propose to face it by recurring to self-organizing maps (SOMs) for structures. Specifically, we show that a suitable combination of the two approaches, obtained by defining a new class of kernels based on the activation map of a SOM for structures, can be effective in avoiding the sparsity problem and results in a system that can be significantly more accurate for categorization tasks on structured data. The effectiveness of the proposed approach is demonstrated experimentally on two relatively large corpora of XML formatted data and a data set of user sessions extracted from website logs.


Explorative Analysis of Wuhan Intra-Urban Human Mobility Using Social Media Check-In Data.
Social media check-in data as a geo-tagged information source have been used for revealing spatio-temporal patterns in the field of social and urban study, such as human behavior or public issues. This paper investigates a case study and presents a new method of representing the mobility of people within a city from check-in data. By dividing the data in a temporal sequence, this study examines the overall mobility in the case study city through the gradient/difference of population density with a series of time after computing the population density from the check-in data using an incorporated Thiessen polygon method. By classifying check-in data with their geo-tags into several groups according to travel purposes, and partitioning the data according to administrative district boundaries, various moving patterns for those travel purposes in those administrative districts are identified by scrutinizing a series of spatial geometries of a weighted standard deviational ellipse (WSDE). Through deep analyses of those data by the adopted approaches, the general pattern of mobility in the case city, such as people moving to the central urban area from the suburb from 4 am to 8 am, is ascertained, and different characteristics of movement in those districts are also depicted. Furthermore, it can tell that in which district less movement is likely for a certain purpose (e.g., for dinner or entertainment). This study has demonstrated the availability of the proposed methodology and check-in data for investigating intra-urban human mobility.


The extended AT-hook is a novel RNA binding motif.
The AT-hook has been defined as a DNA binding peptide motif that contains a glycine-arginine-proline (G-R-P) tripeptide core flanked by basic amino acids. Recent reports documented variations in the sequence of AT-hooks and revealed RNA binding activity of some canonical AT-hooks, suggesting a higher structural and functional variability of this protein domain than previously anticipated. Here we describe the discovery and characterization of the extended AT-hook peptide motif (eAT-hook), in which basic amino acids appear symmetrical mainly at a distance of 12-15 amino acids from the G-R-P core. We identified 80 human and 60 mouse eAT-hook proteins and biochemically characterized the eAT-hooks of Tip5/BAZ2A, PTOV1 and GPBP1. Microscale thermophoresis and electrophoretic mobility shift assays reveal the nucleic acid binding features of this peptide motif, and show that eAT-hooks bind RNA with one order of magnitude higher affinity than DNA. In addition, cellular localization studies suggest a role for the N-terminal eAT-hook of PTOV1 in nucleocytoplasmic shuttling. In summary, our findings classify the eAT-hook as a novel nucleic acid binding motif, which potentially mediates various RNA-dependent cellular processes.


Platelet thromboxane A2 receptors in type I diabetes.
1. Human platelet thromboxane A2 receptor expression on the membrane surface is possibly dynamically regulated by changes either in the ligand concentration or in membrane fluidity. An increased thromboxane A2 production and a decreased membrane fluidity has been reported in diabetic patients. 2. In the present study the binding characteristics of platelet thromboxane A2 receptors have been investigated in nine diabetic patients (type I) and in 15 healthy control subjects by a radioligand-binding method using 9,11-dimethylmethane-11,12-methane-16-[3-125I-4-hydroxyphenyl]-13, 14- dihydro-13-aza-15-tetranor-thromboxane A2 as the radiolabelled ligand. 3. The maximum concentration of binding sites was 163 (SD 35) fmol/10(8) platelets (n = 15) with 987 (SD 209) receptors/platelet in controls, whereas in diabetic patients the maximum concentration of binding sites was 74.2 (SD 28) fmol/10(8) (n = 9) with 447 (SD 172) receptors/platelet (P less than 0.001). The dissociation constants were 18 (SD 4) nmol/l and 21 (SD 6) nmol/l (not significant) in control subjects and in diabetic patients, respectively. Glycated haemoglobin, which is reported to reduce membrane fluidity, was found to be negatively correlated (r = 0.60, P less than 0.05) with thromboxane A2 receptor number in the diabetic patient group. On the contrary, a positive linear correlation between the equilibrium dissociation constant and glycated haemoglobin was found in diabetic patients (r = 0.75, P less than 0.01).


The influence of antigen presentation on the generation of T-helper cells with different functions.
Thymocytes adoptively transferred into syngeneic irradiated recipients can be primed with antigen (KLH) to generate two types of helper function termed 'specific' and 'non-specific'. Low doses of KLH given without adjuvant generate high levels of non-specific compared to 'specific' helper T cells. Large doses of KLH given in adjuvant (FCA) generate high levels of both types of helper T cell. Explantations for this observation are discussed.


Factors affecting routine immunization coverage among children aged 12-59 months in Lao PDR after regional polio eradication in western Pacific region.
The global poliomyelitis eradication programme had a great impact on routine immunization coverage in Lao PDR: DPT3 increased 23% in 1992 to 56% in 1999; OPV3 27% to 64%. However, after the achievement of regional eradication, coverage became stagnant in accordance with the withdrawal of various sources of financial supports. In place of the former funds, a public-private global partnership began to support EPI. We aim to explore factors affecting routine immunization coverage. From February to March of 2005, a cross-sectional questionnaire survey was conducted, targeting 341 mothers living in two districts where immunization coverage was the lowest and the middle in Oudomxay province. DPT3 coverage was 72%, higher than the national target of 65%; however, the drop-out rate was 21%. Influential factors on fully immunized child was distance, literacy, possession of livestock; mothers knowledge of immunization target diseases, measles immunization schedule; and mother's willingness to pay for immunization. In total, 98% of all mothers lived within a 30-minute walk of the immunization site. Household visits increased the immunization status among mothers who were illiterate, utilizing an outreach site for immunization, not willing to pay for immunization, receiving home delivery, and without health education attendance. The much higher routine immunization coverage especially in a district of poor EPI activities suggests a well-designed primary health care approach under the district strategy, the zone-zero social mobilization strategy and good lines of communications; it also points to the benefits of the polio eradication initiative. Household visits were found to be effective for people living with difficulties in such as education, living location, and finance. An equally shared funding system for the basic health as well as international policy for respecting the existing system in poor country is important.


Microbial metabolism and community structure in response to bioelectrochemically enhanced remediation of petroleum hydrocarbon-contaminated soil.
This study demonstrates that electrodes in a bioelectrochemical system (BES) can potentially serve as a nonexhaustible electron acceptor for in situ bioremediation of hydrocarbon contaminated soil. The deployment of BES not only eliminates aeration or supplement of electron acceptors as in contemporary bioremediation but also significantly shortens the remediation period and produces sustainable electricity. More interestingly, the study reveals that microbial metabolism and community structure distinctively respond to the bioelectrochemically enhanced remediation. Tubular BESs with carbon cloth anode (CCA) or biochar anode (BCA) were inserted into raw water saturated soils containing petroleum hydrocarbons for enhancing in situ remediation. Results show that total petroleum hydrocarbon (TPH) removal rate almost doubled in soils close to the anode (63.5-78.7%) than that in the open circuit positive controls (37.6-43.4%) during a period of 64 days. The maximum current density from the BESs ranged from 73 to 86 mA/m(2). Comprehensive microbial and chemical characterizations and statistical analyses show that the residual TPH has a strongly positive correlation with hydrocarbon-degrading microorganisms (HDM) numbers, dehydrogenase activity, and lipase activity and a negative correlation with soil pH, conductivity, and catalase activity. Distinctive microbial communities were identified at the anode, in soil with electrodes, and soil without electrodes. Uncommon electrochemically active bacteria capable of hydrocarbon degradation such as Comamonas testosteroni, Pseudomonas putida, and Ochrobactrum anthropi were selectively enriched on the anode, while hydrocarbon oxidizing bacteria were dominant in soil samples. Results from genus or phylum level characterizations well agree with the data from cluster analysis. Data from this study suggests that a unique constitution of microbial communities may play a key role in BES enhancement of petroleum hydrocarbons biodegradation in soils.


Transcutaneous electrical stimulation of acupoints changes body composition and heart rate variability in postmenopausal women with obesity.
This study aimed to evaluate the effect of transcutaneous electric acupoint stimulations (TEAS) on body composition and heart rate variability (HRV) in postmenopausal women with obesity. In this prospective study, 49 postmenopausal women were recruited in Taiwan. Body composition was used as a screening test for obesity (percentage body fat > 30%, waist circumference > 80 cm). The experimental group (n = 24) received TEAS treatment 30 min twice per week for 12 weeks at the Zusanli (ST 36) and Sanyinjiao (SP 6) acupoints. The control group (n = 25) did not receive any intervention. The study of HRV was analyzed by time (standard deviation of the normal-to-normal (NN) intervals (SDNN) and square root of the mean squared differences of successive NN intervals (RMSSD) indices) and frequency domain methods. Power spectral components were obtained at low (LF) and high (HF) frequencies. Body composition and HRV values were measured at the 4th, 8th, and 12th weeks. A total of 40 subjects completed this study. Waist circumference and percentage body fat in the experimental group (n = 20) were significantly less than those of the control group (n = 20) at the 8th and 12th weeks (all P < .05). Additionally, at the same time points, percentage lean body mass in the experimental group was significantly greater than that in the control group (P < .05). SDNN values increased significantly at the 4th and 8th weeks when compared with the control group (all P < .05). At 12 weeks, SDNN value was not significantly different from that of the control group (P = .105). TEAS treatment improves body composition, and has a transient effect on the HRV in postmenopausal women with obesity.


Dislocation avalanches from strain-controlled loading: A discrete dislocation dynamics study.
We study strain-controlled plastic deformation of crystalline solids via two-dimensional discrete dislocation dynamics simulations. To this end, we characterize the average stress-strain curves as well as the statistical properties of strain bursts and the related stress drops as a function of the imposed strain rate and the stiffness of the specimen-machine system. The dislocation system exhibits strain-rate sensitivity such that a larger imposed strain rate results in a higher average stress at a given strain. In the limit of small strain rate and driving spring stiffness, the sizes and durations of the dislocation avalanches are power law distributed up to a cutoff scale, and exhibit temporally asymmetric average shapes. We discuss the dependence of the results on the driving parameters and compare our results to those from previous simulations where quasistatic stress-controlled loading was used.


Deep subwavelength waveguiding and focusing based on designer surface plasmons.
We experimentally demonstrate focusing and guiding electromagnetic (EM) waves in a designer surface plasmonic waveguide with deep subwavelength mode cross section. Our experiments show that a metal grating with suitable parameters, functioning as a designer surface plasmonic waveguide, can support deep subwavelength surface modes and the width of the modes can be squeezed also into deep subwavelength by tapering the width of the waveguide. The results provide a new insight into deep subwavelength waveguiding and focusing.


A regulatory cascade of three homeobox genes, ceh-10, ttx-3 and ceh-23, controls cell fate specification of a defined interneuron class in C. elegans.
The development of the nervous system requires the coordinated activity of a variety of regulatory factors that define the individual properties of specific neuronal subtypes. We report a regulatory cascade composed of three homeodomain proteins that act to define the properties of a specific interneuron class in the nematode C. elegans. We describe a set of differentiation markers characteristic for the AIY interneuron class and show that the ceh-10 paired-type and ttx-3 LIM-type homeobox genes function to regulate all known subtype-specific features of the AIY interneurons. In contrast, the acquisition of several pan-neuronal features is unaffected in ceh-10 and ttx-3 mutants, suggesting that the activity of these homeobox genes separates pan-neuronal from subtype-specific differentiation programs. The LIM homeobox gene ttx-3 appears to play a central role in regulation of AIY differentiation. Not only are all AIY subtype characteristics lost in ttx-3 mutants, but ectopic misexpression of ttx-3 is also sufficient to induce AIY-like features in a restricted set of neurons. One of the targets of ceh-10 and ttx-3 is a novel type of homeobox gene, ceh-23. We show that ceh-23 is not required for the initial adoption of AIY differentiation characteristics, but instead is required to maintain the expression of one defined AIY differentiation feature. Finally, we demonstrate that the regulatory relationship between ceh-10, ttx-3 and ceh-23 is only partially conserved in other neurons in the nervous system. Our findings illustrate the complexity of transcriptional regulation in the nervous system and provide an example for the intricate interdependence of transcription factor action.


Machine learning methods in sport injury prediction and prevention: a systematic review.
Injuries are common in sports and can have significant physical, psychological and financial consequences. Machine learning (ML) methods could be used to improve injury prediction and allow proper approaches to injury prevention. The aim of our study was therefore to perform a systematic review of ML methods in sport injury prediction and prevention. A search of the PubMed database was performed on March 24th 2020. Eligible articles included original studies investigating the role of ML for sport injury prediction and prevention. Two independent reviewers screened articles, assessed eligibility, risk of bias and extracted data. Methodological quality and risk of bias were determined by the Newcastle-Ottawa Scale. Study quality was evaluated using the GRADE working group methodology. Eleven out of 249 studies met inclusion/exclusion criteria. Different ML methods were used (tree-based ensemble methods (n = 9), Support Vector Machines (n = 4), Artificial Neural Networks (n = 2)). The classification methods were facilitated by preprocessing steps (n = 5) and optimized using over- and undersampling methods (n = 6), hyperparameter tuning (n = 4), feature selection (n = 3) and dimensionality reduction (n = 1). Injury predictive performance ranged from poor (Accuracy = 52%, AUC = 0.52) to strong (AUC = 0.87, f1-score = 85%). Current ML methods can be used to identify athletes at high injury risk and be helpful to detect the most important injury risk factors. Methodological quality of the analyses was sufficient in general, but could be further improved. More effort should be put in the interpretation of the ML models.


Zika virus NS1 structure reveals diversity of electrostatic surfaces among flaviviruses.
The association of Zika virus (ZIKV) infections with microcephaly has resulted in an ongoing public-health emergency. Here we report the crystal structure of a C-terminal fragment of ZIKV nonstructural protein 1 (NS1), a major host-interaction molecule that functions in flaviviral replication, pathogenesis and immune evasion. Comparison with West Nile and dengue virus NS1 structures reveals conserved features but diverse electrostatic characteristics at host-interaction interfaces, thus possibly implying different modes of flavivirus pathogenesis.


'I felt pain. Deep pain…': Experiences of primary caregivers of stroke survivors with aphasia in a South African township.
Aphasia is an acquired impairment in language and in the cognitive processes that underlie language. Aphasia affects the quality of life of the person with aphasia (PWA) and his or her families in various ways in diverse contexts and cultures. It is therefore important that speech language therapists understand how different contextual and cultural factors may mediate experiences. The aim of the study was to describe the caregiving experience of female caregivers of PWA residing in Tembisa, a township situated in the east of Johannesburg. Qualitative, semi-structured interviews were conducted with primary caregivers of PWA. Purposive sampling was used to recruit 14 primary caregivers of PWA who were daughters, daughters-in-law or wives of the PWA. The interviews were conducted in participants' first language and analysed by the researcher, who is proficient in isiZulu. Data were analysed according to the principles of thematic analysis. Findings indicated that caregivers are unfamiliar with aphasia and the support available to them. Participants experienced frustration and found communication to be challenging owing to their lack of communication strategies. The participants' experiences reflected their context-specific experiences, such as feminisation of caregiving, barriers to healthcare, the influence of low health literacy and contextual perspectives on stroke and aphasia. Contextual factors of caregivers in Tembisa have an influence on the experiences between caregivers and PWA, the feelings of individuals and families and health-seeking behaviours of individuals and families.


Who attends antenatal care and expanded programme on immunization services in Chad, Mali and Niger? The implications for insecticide-treated net delivery.
Malaria remains one of the largest public health problems facing the developing world. Insecticide-treated nets (ITNs) are an effective intervention against malaria. ITN delivery through routine health services, such as antenatal care (ANC) and childhood vaccination (EPI), is a promising channel of delivery to reach individuals with the highest risk (pregnant women and children under five years old). Decisions on whether to deliver ITNs through both channels depends upon the reach of each of these systems, whether these are independent and the effectiveness and cost effectiveness of each. Predictors of women attending ANC and EPI separately have been studied, but the predictors of those who attend neither service have not been identified. Data from Chad, Mali and Niger demographic and health surveys (DHS) were analyzed to determine risk factors for attending neither service. A conceptual framework for preventative health care-seeking behaviour was created to illustrate the hierarchical relationships between the potential risk factors. The independence of attending both ANC and EPI was investigated. A multivariate model of predictors for non-attendance was developed using logistic regression. ANC and EPI attendance were found to be strongly associated in all three countries. However, 47% of mothers in Chad, 12% in Mali and 36% in Niger did not attend either ANC or EPI. Region, mother's education and partner's education were predictors of non-attendance in all three countries. Wealth index, ethnicity, and occupation were associated with non-attendance in Mali and Niger. Other predictors included religion, healthcare autonomy, household size and number of children under five. Attendance of ANC and EPI are not independent and therefore the majority of pregnant women in these countries will have the opportunity to receive ITNs through both services. Although attendance at ANC and EPI are not independent, delivery through both systems may still add incrementally to delivery through one alone. Therefore, there is potential to increase the proportion of women and children receiving ITNs by delivering through both of these channels. However, modelling is required to determine the level of attendance and incremental potential at which it's cost effective to deliver through both services.


The gene for incontinentia pigmenti: failure of linkage studies using DNA probes to confirm cytogenetic localization.
Probes for restriction fragment length polymorphisms mapping between Xp21 and Xq22.3 have been used in a linkage study of incontinentia pigmenti (IP). Six independent sporadic cases of disorders resembling IP with X-autosome translocations involving the same X chromosome breakpoint (Xp11) have been reported. These observations suggest that the IP gene may be located in the Xp11 chromosomal region. However, the linkage study with DNA probes has failed to confirm this localisation.


De novo gastric adenocarcinoma 1 year after sleeve gastrectomy in a transplant patient.
It has been reported in the literature that upper gastrointestinal malignancies after bariatric surgery are mostly gastro-esophageal, although it is not clear whether bariatric surgery represents a risk factor for the development of esophageal and/or gastric cancer. We report a case of a de novo gastric adenocarcinoma occurring in a transplant patient 1 year after a laparoscopic sleeve gastrectomy. A 44 year-old woman with a BMI of 38kg/m(2), hypertension, type 1 diabetes mellitus, multiple malignancies and a pancreas transplant underwent laparoscopic sleeve gastrectomy. The patient presented with intense dysphagias during the follow up. Studies were performed and the diagnoses of grade 2/3 adenocarcinoma were made. The patient underwent a robotic assisted total gastrectomy with a roux-en-y intracorporeal esophagojejunostomy. The procedure resulted in multiple metastasic lymph nodes, focal and transmural invasions to multiple organs with a tumor free margin resection. The patient presented with a postoperative pleural effusion, with no further complications. The diagnosis of gastroesophageal cancer after bariatric surgery is usually late since these patients have common upper gastrointestinal symptoms related to the procedure that could delay the diagnosis. De novo gastric cancer after sleeve gastrectomy has only been reported in one instance, in contrast with other bariatric surgery procedures. No direct relation has been established between sleeve gastrectomy and the development of gastric cancer. Robotic procedures allow for complex multiorgan resections, while preserving the benefits of minimally invasive surgery.


Instrumental variable analysis in the presence of unmeasured confounding.
Observational studies are prone to bias due to confounding either measured or unmeasured. While measured confounding can be controlled for with a variety of sophisticated methods such as propensity score-based matching, stratification and multivariable regression model, the unmeasured confounding is usually cumbersome, leading to biased estimates. In econometrics, instrumental variable (IV) is widely used to control for unmeasured confounding. However, its use in clinical researches is generally less employed. In some subspecialties of clinical medicine such as pharmacoepidemiological research, IV analysis is increasingly used in recent years. With the development of electronic healthcare records, more and more healthcare data are available to clinical investigators. Such kind of data are observational in nature, thus estimates based on these data are subject to confounding. This article aims to review several methods for implementing IV analysis for binary and continuous outcomes. R code for these analyses are provided and explained in the main text.


Antibiotic-free production of a herpes simplex virus 2 DNA vaccine in a high yield cGMP process.
Two DNA vaccine plasmids encoding Herpes simplex virus type 2 (HSV-2) glycoprotein D, NTC8485-O2-gD2 and NTC8485-O2-UgD2tr, were produced at large scale under current good manufacturing practice (cGMP) for use in a Phase I human clinical trial. These DNA vaccines incorporate the regulatory agency compliant, minimal, antibiotic-free (AF) NTC8485 mammalian expression vector. Plasmid yields of>1 g/L were achieved using the HyperGRO™ fed-batch fermentation process, with successful scale up from 10 L process development scale to 320 L culture volume for cGMP production. The DNA vaccines were purified using a low residence time, high shear lysis process and AIRMIX(TM) technology, followed by chromatographic purification. This combination of optimized plasmid vector, high yield upstream production, and efficient downstream purification resulted in purified HSV-2 DNA vaccines with>99% total supercoiled plasmid, ≤ 0.2% RNA, ≤ 0.1% host cell genomic DNA, and ≤ 0.1 endotoxin units per mg.


Strong population structure characterizes weediness gene evolution in the invasive grass species Brachypodium distachyon.
Mediterranean annual grasses have invaded California and have replaced vast areas of native grassland. One of these invasive grasses is Brachypodium distachyon, a new model species for the grasses with extensive genomic resources and a nearly completed genome sequence. This study shows that the level of genetic variation in invaded California grasslands is lower compared to the native range in Eurasia. The invaded regions are characterized by highly differentiated populations of B. distachyon isolated by distance, most likely as a result of founder effects and a dearth of outcrossing events. EXP6 and EXP10 encoding alpha-expansins responsible for rapid growth, and AGL11 and AGL13 encoding proteins involved in vegetative phase regulation, appear to be under purifying selection with no evidence for local adaptation. Our data show that B. distachyon has diverged only recently from related Brachypodium species and that tetraploidization might have been as recent as a few thousand years ago. Observed low genetic variation in EXP10 and AGL13 appears to have been present in Eurasia before tetraploidization, potentially as a result of strong selective pressures on advantageous mutations, which are most likely responsible for its fast growth and rapid completion of its life cycle.


Obesity, metabolic factors and risk of different histological types of lung cancer: A Mendelian randomization study.
Assessing the relationship between lung cancer and metabolic conditions is challenging because of the confounding effect of tobacco. Mendelian randomization (MR), or the use of genetic instrumental variables to assess causality, may help to identify the metabolic drivers of lung cancer. We identified genetic instruments for potential metabolic risk factors and evaluated these in relation to risk using 29,266 lung cancer cases (including 11,273 adenocarcinomas, 7,426 squamous cell and 2,664 small cell cases) and 56,450 controls. The MR risk analysis suggested a causal effect of body mass index (BMI) on lung cancer risk for two of the three major histological subtypes, with evidence of a risk increase for squamous cell carcinoma (odds ratio (OR) [95% confidence interval (CI)] = 1.20 [1.01-1.43] and for small cell lung cancer (OR [95%CI] = 1.52 [1.15-2.00]) for each standard deviation (SD) increase in BMI [4.6 kg/m2]), but not for adenocarcinoma (OR [95%CI] = 0.93 [0.79-1.08]) (Pheterogeneity = 4.3x10-3). Additional analysis using a genetic instrument for BMI showed that each SD increase in BMI increased cigarette consumption by 1.27 cigarettes per day (P = 2.1x10-3), providing novel evidence that a genetic susceptibility to obesity influences smoking patterns. There was also evidence that low-density lipoprotein cholesterol was inversely associated with lung cancer overall risk (OR [95%CI] = 0.90 [0.84-0.97] per SD of 38 mg/dl), while fasting insulin was positively associated (OR [95%CI] = 1.63 [1.25-2.13] per SD of 44.4 pmol/l). Sensitivity analyses including a weighted-median approach and MR-Egger test did not detect other pleiotropic effects biasing the main results. Our results are consistent with a causal role of fasting insulin and low-density lipoprotein cholesterol in lung cancer etiology, as well as for BMI in squamous cell and small cell carcinoma. The latter relation may be mediated by a previously unrecognized effect of obesity on smoking behavior.


Development and characterization of a fusion protein between thermally responsive elastin-like polypeptide and interleukin-1 receptor antagonist: sustained release of a local antiinflammatory therapeutic.
Interleukin-1 receptor antagonist (IL-1Ra) has been evaluated for the intraarticular treatment of osteoarthritis. Such administration of proteins may have limited utility because of their rapid clearance and short half-life in the joint. The fusion of a drug to elastin-like polypeptides (ELPs) promotes the formation of aggregating particles that form a "drug depot" at physiologic temperatures, a phenomenon intended to prolong the presence of the drug. The purpose of this study was to develop an injectable drug depot composed of IL-1Ra and ELP domains and to evaluate the properties and bioactivity of the recombinant ELP-IL-1Ra fusion protein. Fusion proteins between IL-1Ra and 2 distinct sequences and molecular weights of ELP were overexpressed in Escherichia coli. Environmental sensitivity was demonstrated by turbidity and dynamic light scattering as a function of temperature. IL-1Ra domain activity was evaluated by surface plasmon resonance, and in vitro antagonism of IL-1-mediated lymphocyte and thymocyte proliferation, as well as IL-1-induced tumor necrosis factor alpha (TNFalpha) expression and matrix metalloproteinase 3 (MMP-3) and ADAMTS-4 messenger RNA expression in human intervertebral disc fibrochondrocytes. IL-1Ra immunoreactivity was assessed before and after proteolytic degradation of the ELP partner. Both fusion proteins underwent supramolecular aggregation at subphysiologic temperatures and slowly resolubilized at 37 degrees C. Interaction with IL-1 receptor was slower in association but equivalent in dissociation as compared with the commercial antagonist. Anti-IL-1 activity was demonstrated by inhibition of lymphocyte and thymocyte proliferation and by decreased TNFalpha expression and ADAMTS-4 and MMP-3 transcription by fibrochondrocytes. ELP domain proteolysis liberated a peptide of comparable size and immunoreactivity as the commercial IL-1Ra. This peptide was more bioactive against lymphocyte proliferation, nearly equivalent to the commercial antagonist. The ELP-IL-1Ra fusion protein proved to retain the characteristic ELP inverse phase-transitioning behavior as well as the bioactivity of the IL-1Ra domain. This technology represents a novel drug carrier designed to prolong the presence of bioactive peptides following intraarticular delivery.