The impact of Aboriginal landscape burning on the Australian biota.
One of the most complex and contentious issues in Australian ecology concerns the environmental impact of Aboriginal landscape burning. This issue is not only important for the development of a comprehensive understanding of the dynamics and evolution of the Australian biota, but is central to the formulation of appropriate strategies for the conservation of the nation's biodiversity. Ethnographic evidence leaves little doubt that Aboriginal burning played a central role in the maintenance of the landscapes subsequently colonized by Europeans. Both 19th century European colonists and anthropologists in the 20th century documented the indispensability of fire as a tool in traditional Aboriginal economies, which have aptly been described as 'fire-stick farming'. Aborigines used fire to achieve short-term outcomes such as providing favourable habitats for herbivores or increasing the local abundance of food plants, but it is not clear whether or not Aborigines had a predictive ecological knowledge of the long-term consequences of their use of fire. A large body of ecological evidence suggests that Aboriginal burning resulted in substantial changes in the geographic range and demographic structure of many vegetation types. Aboriginal burning was important in creating habitat mosaics that favoured the abundance of some mammal species and in the maintenance of infrequently burnt habitats upon which the survival of specialized fauna depends. Aboriginal fire regimes were probably critical for the maintenance of at least one species of tree (Callitris intratropica) in the monsoon tropics. The question of the original impact of humans on the Australian environment is fundamentally speculative because of vague, disputed time frames proposed for the waves of colonization and shifting settlement patterns of Aborigines in the late Quaternary period. There is an inherent circular argument concerning the cause and effect of climate change, vegetation change, and burning through the late Quaternary. Charcoal and pollen evidence from long sedimentary cores is ambiguous and cannot be used to demonstrate unequivocally the initial impact of Aboriginal people on the landscapes of Pleistocene Australia. The sparse available evidence does not support the hypotheses that Aboriginal burning was primarily responsible for the extinction of Pleistocene megafauna; was critical for the maintenance of habitats of small mammals that have become extinct following European colonization; initiated widespread accelerated soil erosion rates in either the Pleistocene or Holocene; or forced the evolutionary diversification of the Australian biota. Burning may have caused the extinction of some fire-sensitive species of plants and animals dependent upon infrequently burnt habitats, and it must have maintained structurally open vegetation such as grasslands and also extended the range of fire-adapted species, such as Eucalyptus, into environments climatically suitable for rain forest. Palaeoecological research concerning prior impacts of Aborigines must give way to focused studies of the role of different anthropogenic fire regimes in contemporary ecosystems that have not been destroyed by European colonization. Such research is crucial for comprehending the role of Aboriginal burning in the maintenance of Australia's unique, rich biodiversity. CONTENTS Summary 385 I. Introduction 386 II. Aborigines and fire 386 III. Ecological perspectives 390 IV. Palaeoecological perspectives 394 V. General conclusions 404 Acknowledgements 405 References 405.


Animal Models of Maternal Immune Activation in Depression Research.
Depression and schizophrenia are debilitating mental illnesses with significant socio-economic impact. The high degree of comorbidity between the two disorders, and shared symptoms and risk factors, suggest partly common pathogenic mechanisms. Supported by human and animal studies, maternal immune activation (MIA) has been intimately associated with the development of schizophrenia. However, the link between MIA and depression has remained less clear, in part due to the lack of appropriate animal models. Here we aim to summarize findings obtained from studies using MIA animal models and discuss their relevance for preclinical depression research. Results on molecular, cellular and behavioral phenotypes in MIA animal models were collected by literature search (PubMed) and evaluated for their significance for depression. Several reports on offspring depression-related behavioral alterations indicate an involvement of MIA in the development of depression later in life. Depression-related behavioral phenotypes were frequently paralleled by neurogenic and neurotrophic deficits and modulated by several genetic and environmental factors. Literature evidence analyzed in this review supports a relevance of MIA as animal model for a specific early life adversity, which may prime an individual for the development of distinct psychopathologies later life. MIA animal models may present a unique tool for the identification of additional exogenous and endogenous factors, which are required for the manifestation of a specific neuropsychiatric disorder, such as depression, later in life. Hereby, novel insights into the molecular mechanisms involved in the pathophysiology of depression may be obtained, supporting the identification of alternative therapeutic strategies.


Prospective comparison of breast pain in patients participating in a randomized trial of breast-conserving surgery and tamoxifen with or without radiotherapy.
To determine whether breast pain affects quality of life (QOL) after breast-conserving surgery and tamoxifen (TAM) with or without adjuvant breast radiotherapy (RT). A randomized clinical trial was carried out at the Princess Margaret Hospital between 1992 and 2000 to evaluate the need for breast RT in addition to TAM in women >or=50 years treated with breast-conserving surgery for T1-T2N0 breast cancer. A companion study to assess breast pain was carried out during the last 2 years of the randomized clinical trial. The short-form McGill Pain Questionnaire (SF-MPQ), the European Organization for Research and Treatment of Cancer (EORTC) QOL (QLQ-C30) and EORTC breast cancer module (QLQ-BR23) questionnaires were completed by patients within 1 week of randomization in the randomized clinical trial (baseline) and at 3, 6, and 12 months. Eighty-six patients participated in the breast pain study; 41 received RT plus TAM and 45 received TAM alone. The median age was 70 years (range 51-80). The baseline pain and QOL scores were similar for the two groups. No significant difference was found between the two groups for each scale of the QLQ-C30 and QLQ-BR23 questionnaires at 3, 6, or 12 months (p >0.100), except that at 12 months, the score for role function (QLQ-C30) was higher in the RT plus TAM group than in the RT-only group (p = 0.02). At 3 months, the difference between the mean scores for the SF-MPQ was 0.553 (p = 0.47). At 12 months, the pain scores had decreased in both groups; the difference was 0.199 (p = 0.71). The number of breast operations or surgical complications did not correlate with breast pain in either group. Acute RT toxicity scores did not correlate with breast pain or QOL scores at 12 months. These results suggest that breast RT does not significantly contribute to breast pain or adversely impact the QOL up to 12 months after treatment in postmenopausal patients with node-negative breast cancer who take TAM.


Synthesis of magnetic molecularly imprinted nanoparticles with multiple recognition sites for the simultaneous and selective capture of two glycoproteins.
Many diseases ordinarily have several kinds of biomarkers (most of them belong to the glycoprotein family). Simultaneous capturing of multiple target glycoprotein biomarkers can obtain better specificity compared with single marker detection in early screening of many diseases. In this work, we attempted to prepare magnetic molecularly imprinted nanoparticles with multiple recognition sites. Horseradish peroxidase and ovalbumin as the frequently used glycoproteins for simulation of glycoprotein biomarkers were employed as dual-templates. The simultaneous preorganization of two templates was achieved by introducing 4-formylphenylboronic acid as a functional monomer. To gain a high adsorption capacity for the two templates, the mass ratio of the two templates was precisely investigated. Controllable polymerization of dopamine was applied to synthesize thin imprinted layers that reduce mass transfer resistance and improve the eluted efficiency of the two templates. The resultant magnetic molecularly imprinted nanoparticles exhibited high adsorption capacities of 34.09 and 51.18 mg g-1, respectively. The simultaneous recognition of two templates in spiked fetal bovine serum was also achieved with satisfactory selectivity. The results indicate that this proposed strategy is potentially applicable for simultaneously and selectively capturing several glycoprotein biomarkers used in detecting many diseases.


Inventing conflicts of interest: a history of tobacco industry tactics.
Confronted by compelling peer-reviewed scientific evidence of the harms of smoking, the tobacco industry, beginning in the 1950s, used sophisticated public relations approaches to undermine and distort the emerging science. The industry campaign worked to create a scientific controversy through a program that depended on the creation of industry-academic conflicts of interest. This strategy of producing scientific uncertainty undercut public health efforts and regulatory interventions designed to reduce the harms of smoking. A number of industries have subsequently followed this approach to disrupting normative science. Claims of scientific uncertainty and lack of proof also lead to the assertion of individual responsibility for industrially produced health risks.


Effect of leukocyte-reduced platelet-rich plasma on osteoarthritis caused by cranial cruciate ligament rupture: A canine gait analysis model.
The goal of this study was to objectively assess the effect of a platelet-rich plasma (PRP) derivate in English bulldogs with stifle degenerative joint disease secondary to cranial cruciate ligament rupture (CCLR). We used a force platform and affixed electrogoniometers to measure peak vertical force (PVF), vertical impulse (VI), stance time (ST), and angular range of motion (AROM), from 12 lame client-owned English bulldogs with post-CCLR stifle joint abnormalities. The 12 affected subjects were treated with 4 intra-articular injections of PRP, at 30-day intervals. Ten untreated, sound English bulldogs were used as a reference group. Clinical outcomes were evaluated using a linear mixed effects model. Mean values of PVF, VI, ST, and AROM were improved within the first 3 months post-treatment in the CCLR group, with mean measured changes increasing to maximum 4.56% body weight gain, 1.5% body weight/second, 0.07 seconds, and 6.18 degrees, respectively. The effects declined progressively after the treatment interval, ending at nearly initial levels after 6 months. This study demonstrates that dogs with CCLR treated with intra-articular PRP had improved PVF, VI, ST, and AROM over time; the duration of effect was waning by the end of the post-treatment period.


Superficial amygdala and hippocampal activity during affective music listening observed at 3 T but not 1.5 T fMRI.
The purpose of this study was to compare 3 T and 1.5 T fMRI results during emotional music listening. Stimuli comprised of psychoacoustically balanced instrumental musical pieces, with three different affective expressions (fear, neutral, joy). Participants (N=32) were split into two groups, one subjected to fMRI scanning using 3 T and another group scanned using 1.5 T. Whole brain t-tests (corrected for multiple comparisons) compared joy and fear in each of the two groups. The 3 T group showed significant activity differences between joy and fear localized in bilateral superficial amygdala, bilateral hippocampus and bilateral auditory cortex. The 1.5 T group showed significant activity differences between joy and fear localized in bilateral auditory cortex and cuneus. This is the first study to compare results obtained under different field strengths with regard to affective processes elicited by means of auditory/musical stimulation. The findings raise concern over false negatives in the superficial amygdala and hippocampus in affective studies conducted under 1.5 T and caution that imaging improvements due to increasing magnetic field strength can be influenced by region-specific characteristics.


Using a Bayesian modelling approach (INLA-SPDE) to predict the occurrence of the Spinetail Devil Ray (Mobular mobular).
To protect the most vulnerable marine species it is essential to have an understanding of their spatiotemporal distributions. In recent decades, Bayesian statistics have been successfully used to quantify uncertainty surrounding identified areas of interest for bycatch species. However, conventional simulation-based approaches are often computationally intensive. To address this issue, in this study, an alternative Bayesian approach (Integrated Nested Laplace Approximation with Stochastic Partial Differential Equation, INLA-SPDE) is used to predict the occurrence of Mobula mobular species in the eastern Pacific Ocean (EPO). Specifically, a Generalized Additive Model is implemented to analyze data from the Inter-American Tropical Tuna Commission's (IATTC) tropical tuna purse-seine fishery observer bycatch database (2005-2015). The INLA-SPDE approach had the potential to predict both the areas of importance in the EPO, that are already known for this species, and the more marginal hotspots, such as the Gulf of California and the Equatorial area which are not identified using other habitat models. Some drawbacks were identified with the INLA-SPDE database, including the difficulties of dealing with categorical variables and triangulating effectively to analyze spatial data. Despite these challenges, we conclude that INLA approach method is an useful complementary and/or alternative approach to traditional ones when modeling bycatch data to inform accurately management decisions.


Simian virus 40 (SV40) DNA replication: SV40 large T antigen unwinds DNA containing the SV40 origin of replication.
The simian virus 40 (SV40) large T antigen (large tumor antigen), in conjunction with a topoisomerase, a DNA binding protein, and ATP, catalyzed the conversion of a circular duplex DNA molecule containing the SV40 origin of replication to a form with unusual electrophoretic mobility that we have named form U. Analysis of this molecule revealed it to be a highly underwound covalently closed circle. DNA unwinding was not detected with DNA containing a SV40 T-antigen binding site II mutation that renders the DNA inactive in replication. The unwinding reaction requires the action of a helicase, and SV40 T-antigen preparations contain such an activity. The T-antigen-associated ability to unwind DNA copurified with other activities intrinsic to T antigen [ability to support replication of SV40 DNA containing the SV40 origin, poly(dT)-stimulated ATPase activity, and DNA helicase]. However, in contrast to the unwinding activity, the SV40 T-antigen-associated helicase activity was not sequence-specific. A variety of labeled oligonucleotides hybridized with circular single-stranded DNA were displaced by T antigen in the presence of ATP.


Deep-Learned Regularization and Proximal Operator for Image Compressive Sensing.
Deep learning has recently been intensively studied in the context of image compressive sensing (CS) to discover and represent complicated image structures. These approaches, however, either suffer from nonflexibility for an arbitrary sampling ratio or lack an explicit deep-learned regularization term. This paper aims to solve the CS reconstruction problem by combining the deep-learned regularization term and proximal operator. We first introduce a regularization term using a carefully designed residual-regressive net, which can measure the distance between a corrupted image and a clean image set and accurately identify to which subspace the corrupted image belongs. We then address a proximal operator with a tailored dilated residual channel attention net, which enables the learned proximal operator to map the distorted image into the clean image set. We adopt an adaptive proximal selection strategy to embed the network into the loop of the CS image reconstruction algorithm. Moreover, a self-ensemble strategy is presented to improve CS recovery performance. We further utilize state evolution to analyze the effectiveness of the designed networks. Extensive experiments also demonstrate that our method can yield superior accurate reconstruction (PSNR gain over 1 dB) compared to other competing approaches while achieving the current state-of-the-art image CS reconstruction performance. The test code is available at https://github.com/zjut-gwl/CSDRCANet.


Approaches to studying the transmission of Mycobacterium leprae.
A collaborative study has been undertaken to establish the relationship between infection by Mycobacterium leprae and the development of immunity in a community in which multidrug therapy (MDT) has been used for more than 10 years, to elucidate the pathogenesis of infection in leprosy, and to develop and test an intervention strategy based on chemotherapy for interruption of transmission of the organism in the community. The first phase of the study included the establishment of laboratory facilities and pilot work in India. In the course of the second phase, the entire populations of three villages in India and one in Ethiopia have been surveyed, nasal swabs were obtained for detection of M. leprae DNA by means of the polymerase chain reaction (PCR), specimens of saliva were obtained for measurement of levels of anti-M. leprae IgA antibodies, and follow-up surveys have been carried out. A double-blind trial of chemotherapy among subjects whose PCR was positive is proposed, to determine if the course of the infection can be influenced by treatment. The performance of large numbers of PCR tests in endemic countries has required the development of rigorous internal and external quality control procedures. These have shown that many batches (as many as 50%) fail to meet quality control criteria, and must be retested. Despite this, development of these methods, and their application to field studies should provide tools for studying the transmission of M. leprae, and direct methods of testing innovative interventions.


Symmetries of hydrogen bonds in solution.
The nuclear magnetic resonance method of isotopic perturbation can distinguish between single- and double-well potentials in intramolecularly hydrogen-bonded monoanions of dicarboxylic acids. These are classic cases of a "strong," symmetric hydrogen bond in the crystal. The observed carbon-13 isotope shifts induced by the substitution of oxygen-18 demonstrate that these monoanions exist as a single symmetric structure in a nonpolar solvent but as two equilibrating tautomers in aqueous solution. The change is attributed to the disorder of the aqueous environment. These are simple counterexamples to the hope that the crystal structure reveals the actual molecular structure in aqueous solution.


Web-based, participant-driven studies yield novel genetic associations for common traits.
Despite the recent rapid growth in genome-wide data, much of human variation remains entirely unexplained. A significant challenge in the pursuit of the genetic basis for variation in common human traits is the efficient, coordinated collection of genotype and phenotype data. We have developed a novel research framework that facilitates the parallel study of a wide assortment of traits within a single cohort. The approach takes advantage of the interactivity of the Web both to gather data and to present genetic information to research participants, while taking care to correct for the population structure inherent to this study design. Here we report initial results from a participant-driven study of 22 traits. Replications of associations (in the genes OCA2, HERC2, SLC45A2, SLC24A4, IRF4, TYR, TYRP1, ASIP, and MC1R) for hair color, eye color, and freckling validate the Web-based, self-reporting paradigm. The identification of novel associations for hair morphology (rs17646946, near TCHH; rs7349332, near WNT10A; and rs1556547, near OFCC1), freckling (rs2153271, in BNC2), the ability to smell the methanethiol produced after eating asparagus (rs4481887, near OR2M7), and photic sneeze reflex (rs10427255, near ZEB2, and rs11856995, near NR2F2) illustrates the power of the approach.


A trial of the effect of a standardized psychiatric consultation on health outcomes and costs in somatizing patients.
Patients who somatize but who do not meet criteria for somatization disorder are common in the community. Virtually no research has been conducted to determine how to treat these patients. We conducted a randomized controlled clinical trial of a psychiatric consultation intervention we had previously shown to improve the management of somatization disorder. The study population included 51 physicians treating 56 somatizing patients who had a history of seeking help for six to 12 lifetime unexplained physical symptoms. At the onset of the experiment, physicians randomized to the treatment condition received a consultation letter recommending a specific management approach; physicians randomized to the control/crossover condition received the consultation letter after 12 months. Data on health outcomes and charges were collected every 4 months for 2 years after randomization for 96% of subjects who entered the study. Patients of physicians who received the intervention reported significantly increased physical functioning, an improvement that remained stable during the year after the intervention. The intervention reduced annual medical care charges by $289 (95% confidence interval, $40 to $464) in 1990 constant dollars, which equates to a 32.9% reduction in the annual median cost of their medical care. Somatizing patients with a lifetime history of six to 12 unexplained physical symptoms reported better physical functioning after their primary care physician was provided appropriate treatment recommendations via a psychiatric consultation. Such a consultation is cost-effective because it reduces subsequent charges for medical care, while improving health outcomes in a chronically impaired population.


Endoscopic features and prognoses of mantle cell lymphoma with gastrointestinal involvement.
To evaluate the endoscopic manifestations and prognoses of gastrointestinal (GI) mantle cell lymphoma (MCL). A database search at the Department of Pathology of Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences revealed 57 MCL patients with GI involvement. Clinical records were available for 35 of the 57 patients from 21 institutions, and those 35 patients were enrolled in this study. We summarized the gross types of endoscopic features, event-free survival (EFS), and overall survival (OS) of those patients. Of the 35 patients, GI involvement in the esophagus, stomach, and duodenum was found in 2 (5.7%), 26 (74.3%), and 12 (34.3%) patients, respectively. Twenty-one of the 35 patients underwent colonoscopy; among them, GI involvement in the ileum, cecum, colon, and rectum was found in 10 (47.6%), 3 (14.3%), 12 (57.1%), and 10 (47.6%), respectively. Various lesions, such as superficial, protruded, fold thickening, or ulcerative, were found in the stomach, whereas multiple lymphomatous polyposis (MLP) was dominant from the duodenum to the rectum. Twelve patients were treated with a hyper-CVAD/MA regimen, and they had better OS (3-year rate, 88.3% vs 46.4%, P < 0.01) and better EFS (3-year rate, 66.7% vs 33.8%, P < 0.05) than the remaining 23 patients who were not treated with this regimen. MLP was a representative form of intestinal involvement, whereas a variety of lesions were found in the stomach. The hyper-CVAD/MA regimen may improve survival in these patients.


Inherited disorders of the neuromuscular junction: an update.
Congenital myasthenic syndromes (CMSs) are a group of heterogeneous inherited disorders caused by mutations in genes affecting the function and structure of the neuromuscular junction. This review updates the reader on established and novel subtypes of congenital myasthenia, and the treatment strategies for these increasingly heterogeneous disorders. The discovery of mutations associated with the N-glycosylation pathway and in the family of serine peptidases has shown that causative genes encoding ubiquitously expressed molecules can produce defects at the human neuromuscular junction. By contrast, mutations in lipoprotein-like receptor 4 (LRP4), a long-time candidate gene for congenital myasthenia, and a novel phenotype of myasthenia with distal weakness and atrophy due to mutations in AGRN have now been described. In addition, a pathogenic splicing mutation in a nonfunctional exon of CHRNA1 has been reported emphasizing the importance of analysing nonfunctional exons in genetic analysis. The benefit of salbutamol and ephedrine alone or combined with pyridostigmine or 3,4-DAP is increasingly being reported for particular subtypes of CMS.


The gonads of human true hermaphrodites.
Gonadal distribution in 409 cases of human true hermaphroditism is reviewed. An ovary was found on the left side of the body in 62.8% of the cases and the testis on the right side in 59.5%. The ovotestis is the most common gonad of the true hermaphrodite; amongst 806 gonads in 406 cases it was found in 44.3%. In this paper we give a detailed description of the morphology of ovotestis, testis and ovary in the true hermaphrodite. In addition we discuss the effects of fetal androgens and Müllerian inhibiting factor on the Wolffian and Müllerian ducts. Correlations between chromosomal complement and gonadal distribution are presented. True hermaphrodites with a 46,XX karyotype most commonly have an ovary on one side and an ovotestis on the other side; those with a Y-chromosome have a testis in 61% of cases. An analysis of the ratio of ovarian and testicular tissue within ovotestes showed a continuum from very little ovarian tissue to a small portion of testicular tissue. Each type of tissue was clearly demarcated. Hypotheses for gonadal induction in the true hermaphrodite should take cognizance of these facts. True hermaphrodites with a 46,XX chromosomal complement were characterized by a male phenotype in 54% of cases. This group may suggest a greater testicular induction ability in the genome as compared to the 46% with a female phenotype.


Phosphorylation and activation of 13S condensin by Cdc2 in vitro.
13S condensin is a multisubunit protein complex essential for mitotic chromosome condensation in Xenopus egg extracts. Purified 13S condensin introduces positive supercoils into DNA in the presence of topoisomerase I and adenosine triphosphate in vitro. The supercoiling activity of 13Scondensin was regulated by mitosis-specific phosphorylation. Immunodepletion, in vitro phosphorylation, and peptide-mapping experiments indicated that Cdc2 is likely to be the kinase that phosphorylates and activates 13S condensin. Multiple Cdc2 phosphorylation sites are clustered in the carboxyl-terminal domain of the XCAP-D2 (Xenopus chromosome-associated polypeptide D2) subunit. These results suggest that phosphorylation of 13Scondensin by Cdc2 may trigger mitotic chromosome condensation in vitro.


Poly(Ethylene glycol)-based backbones with high peptide loading capacities.
Polymer-peptide conjugates are a promising class of compounds, where polymers can be used to overcome some of the limitations associated with peptides intended for therapeutic and/or diagnostic applications. Linear polymers such as poly(ethylene glycol) can be conjugated through terminal moieties and have therefore limited loading capacities. In this research, functionalised linear poly(ethylene glycol)s are utilised for peptide conjugation, to increase their potential loading capacities. These poly(ethylene glycol) derivatives are conjugated to peptide sequences containing representative side-chain functionalised amino acids, using different conjugation chemistries, including copper-catalysed azide-alkyne cycloaddition, amide coupling and thiol-ene reactions. Conjugation of a sequence containing the RGD motif to poly(allyl glycidyl ether) by the thiol-ene reaction, provided a conjugate which could be used in platelet adhesion studies.


Subacute spinal subdural hematoma after spontaneous resolution of cranial subdural hematoma: causal relationship or coincidence? Case report.
The etiopathogenesis of traumatic spinal subdural hematoma (SSH) is uncertain. Unlike the supratentorial subdural space, no bridging veins traverse the spinal subdural space. The authors describe a case of subacute SSH that occurred after spontaneous resolution of traumatic intracranial SDH and suggest a causal relationship between the two. A 23-year-old woman suffered an acute intracranial SDH after a snowboarding accident. There was no clinical or radiological evidence of spine injury. Conservative management of the supratentorial SDH resulted in spontaneous radiologically documented resolution with redistribution of blood in the subdural space. Four days after the injury, the patient started noticing new onset of mild low-back pain. The pain progressively worsened. Magnetic resonance imaging of the lumbosacral spine 10 days after the original injury revealed a large L4-S2 SDH. Ten days after the original injury, bilateral L5-S1 laminotomy and drainage of the subacute spinal SDH were performed. The patient experienced immediate pain relief. The authors hypothesize that in some cases spinal SDH may be related to redistribution of blood from the supratentorial subdural space.


Functional tissue engineering: the role of biomechanics in articular cartilage repair.
Articular cartilage shows little or no intrinsic capacity for repair in response to injury or disease, and even minor lesions or injuries may lead to progressive damage and joint degeneration. Tissue engineering is a relatively new but rapidly growing field that has sought to use combinations of implanted cells, biomaterials, and biologically active molecules to repair or regenerate injured or diseased tissues. Despite many advances, tissue engineers have faced significant challenges in repairing or replacing tissues that serve a predominantly biomechanical function, such as articular cartilage. An evolving discipline termed functional tissue engineering seeks to address these challenges by emphasizing and evaluating the role of biomechanical factors in the intrinsic and engineered repair of tissues and organs. In the current study, the authors describe some of the fundamental issues involving the interaction of biomechanical stresses in vivo and in vitro with native and repair articular cartilage and with other biomechanically functional tissues. A more thorough and formal investigation of these issues may provide a basis for developing rational design principles for tissue engineered replacement or repair of load-bearing structures in the body.


Redistribution of flexibility in stabilizing antibody fragment mutants follows Le Châtelier's principle.
Le Châtelier's principle is the cornerstone of our understanding of chemical equilibria. When a system at equilibrium undergoes a change in concentration or thermodynamic state (i.e., temperature, pressure, etc.), La Châtelier's principle states that an equilibrium shift will occur to offset the perturbation and a new equilibrium is established. We demonstrate that the effects of stabilizing mutations on the rigidity ⇔ flexibility equilibrium within the native state ensemble manifest themselves through enthalpy-entropy compensation as the protein structure adjusts to restore the global balance between the two. Specifically, we characterize the effects of mutation to single chain fragments of the anti-lymphotoxin-β receptor antibody using a computational Distance Constraint Model. Statistically significant changes in the distribution of both rigidity and flexibility within the molecular structure is typically observed, where the local perturbations often lead to distal shifts in flexibility and rigidity profiles. Nevertheless, the net gain or loss in flexibility of individual mutants can be skewed. Despite all mutants being exclusively stabilizing in this dataset, increased flexibility is slightly more common than increased rigidity. Mechanistically the redistribution of flexibility is largely controlled by changes in the H-bond network. For example, a stabilizing mutation can induce an increase in rigidity locally due to the formation of new H-bonds, and simultaneously break H-bonds elsewhere leading to increased flexibility distant from the mutation site via Le Châtelier. Increased flexibility within the VH β4/β5 loop is a noteworthy illustration of this long-range effect.


Risk factors and biomarkers of life-threatening cancers.
There is growing evidence that risk factors for cancer occurrence and for cancer death are not necessarily the same. Knowledge of cancer aggressiveness risk factors (CARF) may help in identifying subjects at high risk of developing a potentially deadly cancer (and not just any cancer). The availability of CARFs may have positive consequences for health policies, medical practice, and the search for biomarkers. For instance, cancer chemoprevention and cancer screening of subjects with CARFs would probably be more ethical and cost-effective than recommending chemoprevention and screening to entire segments of the population. Also, the harmful consequences of chemoprevention and of screening would be reduced while effectiveness would be optimised. We present examples of CARF already in use (e.g. mutations of the breast cancer (BRCA) gene), of promising avenues for the discovery of biomarkers thanks to the investigation of CARFs (e.g. breast radiological density and systemic inflammation), and of biomarkers commonly used that are not real CARFs (e.g. certain mammography images, prostate-specific antigen (PSA) concentration, nevus number).


Relationship of Smokefree Laws and Alcohol Use with Light and Intermittent Smoking and Quit Attempts among US Adults and Alcohol Users.
Light and intermittent smoking (LITS) has become increasingly common. Alcohol drinkers are more likely to smoke. We examined the association of smokefree law and bar law coverage and alcohol use with current smoking, LITS, and smoking quit attempts among US adults and alcohol drinkers. Cross-sectional analyses among a population-based sample of US adults (n = 27,731) using restricted data from 2009 National Health Interview Survey and 2009 American Nonsmokers' Rights Foundation United States Tobacco Control Database. Multivariate logistic regression models examined the relationship of smokefree law coverage and drinking frequency (1) with current smoking among all adults; (2) with 4 LITS patterns among current smokers; and (3) with smoking quit attempts among 6 smoking subgroups. Same multivariate analyses were conducted but substituted smokefree bar law coverage for smokefree law coverage to investigate the association between smokefree bar laws and the outcomes. Finally we ran the above analyses among alcohol drinkers (n = 16,961) to examine the relationship of smokefree law (and bar law) coverage and binge drinking with the outcomes. All models controlled for demographics and average cigarette price per pack. The interactions of smokefree law (and bar law) coverage and drinking status was examined. Stronger smokefree law (and bar law) coverage was associated with lower odds of current smoking among all adults and among drinkers, and had the same effect across all drinking and binge drinking subgroups. Increased drinking frequency and binge drinking were related to higher odds of current smoking. Smokefree law (and bar law) coverage and drinking status were not associated with any LITS measures or smoking quit attempts. Stronger smokefree laws and bar laws are associated with lower smoking rates across all drinking subgroups, which provides further support for these policies. More strict tobacco control measures might help reduce cigarette consumption and increase quit attempts.


Communicating uncertainties about prescription drugs to the public: a national randomized trial.
Many new drugs are aggressively promoted. The public may not realize that even with US Food and Drug Administration (FDA) approval, important uncertainties about the benefits and harms of these drugs remain. We assessed the US public's understanding of the meaning of FDA drug approval and tested how brief explanations communicating drug uncertainties affect consumer choices. We conducted an Internet-based randomized controlled trial using a national sample of US adults from a research panel of approximately 30,000 households. A total of 2944 participants were randomized to receive 1 of 3 explanations about a pair of cholesterol drugs (1 approved based only on a surrogate outcome [lower cholesterol] and 1 based on a patient outcome [reduced myocardial infarctions]). Participants were randomized a second time to receive 1 of 3 explanations about a pair of heartburn drugs (1 newly approved and 1 approved 8 years earlier). Controls received no explanation; the nondirective group received explanations (for the cholesterol drugs, surrogates do not always translate into patient outcomes; for the heartburn drugs, it takes time to establish the safety of new drugs); the directive group received explanations plus advice to "Ask for a drug shown to reduce heart attacks or ask for one with a longer track record." The primary outcomes were choice: the cholesterol drug reducing myocardial infarctions, and the older heartburn drug. Thirty-nine percent mistakenly believed that the FDA approves only "extremely effective" drugs; 25% mistakenly believed that the FDA approves only drugs without serious side effects. Explanations affected choices: 71% of those in the directive group, 71% in the nondirective group, and 59% of controls chose the cholesterol drug that reduced myocardial infarctions (absolute difference, 12% [95% confidence interval, 7%-18%] for each explanation vs control). For the heartburn drugs, 53% of the directive group, 53% of the nondirective group, and 34% of controls chose the older drug (absolute difference, 19% [95% confidence interval, 13%-24%] for each explanation vs control). A substantial proportion of the public mistakenly believes that the FDA approves only extremely effective drugs and drugs lacking serious side effects. Brief explanations highlighting uncertainties about the benefit of drugs approved based on surrogate outcomes and the safety of new prescription drugs improved choices. Nondirective explanations worked as well as directive ones. clinicaltrials.gov Identifiers: NCT00950157, NCT00950131.


Frailty status and altered dynamics of circulating energy metabolism hormones after oral glucose in older women.
Frailty status is associated with altered glucose-insulin dynamics. Here, we sought to investigate whether alteration in the dynamics of other circulating energy metabolism hormones after oral glucose is associated with frailty status. Substudy of older women in a prospective cohort. Baltimore, Maryland. Seventy-three community-dwelling women aged 84-95 years without a diagnosis of diabetes who were enrolled in the Women's Health and Aging Study II. We examined stimulus-response dynamics of free fatty acids (FFA), gut- (ghrelin,GLP-1) and adipocyte-derived hormones (leptin, adiponectin, resistin), growth hormone (GH), insulin-like growth factor 1 (IGF-1), and interleukin-6 (IL-6) at 0, 30, 60, 120, and 180-minutes after a 75-g glucose challenge according to frailty status (non-frail, pre-frail, or frail). On average, frail women had higher fasting levels of glucose-raising hormones (FFA, resistin, GH, and IL-6) and lower fasting levels of glucose-lowering hormones (ghrelin, adiponectin, GLP-1 and IGF-1) versus non-frail women but these results were not statistically significant. Frail women also had higher fasting levels of leptin with relative adiposity compared to their counterparts, suggestive of leptin-resistance. Integrated area under the curve (AUC) values for each hormone followed similar trends by frailty status. After age and BMI adjustment, frail versus non-frail women were more likely to be in the lowest tertile of fasting ghrelin levels and 120-min ghrelin levels (both p<0.05) in logistic regression analyses. No large differences were found for other hormones in adjusted models. Our findings suggest dysregulation of the orexigenic hormone ghrelin in the frailty syndrome. Further studies are needed to explore the role of ghrelin dysregulation in the clinical manifestation of frailty.


Hepatitis C virus infection and biological false-positive syphilis tests.
The diagnosis of syphilis requires two-step serological testing. Not infrequently, sensitive screening tests are reactive but are not confirmed by more specific confirmatory tests yielding a biological false positive (BFP). This study sought to describe the prevalence of BFP in a large population of hepatitis C virus (HCV)-infected and uninfected women. A cross-sectional serosurvey of HIV-seropositive and HIV-seronegative women enrolled in the Women's Interagency HIV Study, a multicentre collaborative study of the natural history of HIV in women. Among HCV-infected women 4% had a BFP compared with 1% among those who were HCV uninfected (odds ratio (OR) 3.3, 95% CI 2.1 to 5.1). Controlling for both HIV infection and a history of intravenous drug use among all tests for syphilis a BFP also occurred more commonly in HCV-infected women compared with HCV-uninfected women (6% vs 1%, OR 7.62, 95% CI 1.9 to 12.5). HCV infection is associated with various effects on immune function including alterations in serological test results. Women with HCV are more likely to have a BFP syphilis test than women without HCV.


Integral Membrane Protein 2A Is a Negative Regulator of Canonical and Non-Canonical Hedgehog Signalling.
The Hedgehog (Hh) receptor PTCH1 and the integral membrane protein 2A (ITM2A) inhibit autophagy by reducing autolysosome formation. In this study, we demonstrate that ITM2A physically interacts with PTCH1; however, the two proteins inhibit autophagic flux independently, since silencing of ITM2A did not prevent the accumulation of LC3BII and p62 in PTCH1-overexpressing cells, suggesting that they provide alternative modes to limit autophagy. Knockdown of ITM2A potentiated PTCH1-induced autophagic flux blockade and increased PTCH1 expression, while ITM2A overexpression reduced PTCH1 protein levels, indicating that it is a negative regulator of PTCH1 non-canonical signalling. Our study also revealed that endogenous ITM2A is necessary for timely induction of myogenic differentiation markers in C2C12 cells since partial knockdown delays the timing of differentiation. We also found that basal autophagic flux decreases during myogenic differentiation at the same time that ITM2A expression increases. Given that canonical Hh signalling prevents myogenic differentiation, we investigated the effect of ITM2A on canonical Hh signalling using GLI-luciferase assays. Our findings demonstrate that ITM2A is a strong negative regulator of GLI transcriptional activity and of GLI1 stability. In summary, ITM2A negatively regulates canonical and non-canonical Hh signalling.


Autologous mixed lymphocyte reaction in the peripheral blood and pleural effusions of cancer patients.
T cells proliferate in response to autologous non-T cells in the autologous mixed lymphocyte reaction (AMLR). AMLR was impaired in the peripheral blood of patients with advanced lung cancer (4,159 +/- 3,878 delta cpm vs. 11,221 +/- 4,156 delta cpm for normal donors) but normal or even higher in their malignant pleural effusions (13,257 +/- 7,075 delta cpm vs. 10, 870 +/- 5,013 delta cpm for nonmalignant control effusions). Blood T cells also failed to respond to autologous effusion non-T cells, while effusion T cells strongly responded to autologous erythrocytes blood non-T cells. The presence of blood T cells did not inhibit effusion AMLR of the same patients. A subset of T cells that form rosettes with autologous erythrocytes if found to proliferate in AMLR. The number of autorosette-forming cells was lower in blood T cells of cancer patients than in blood T cells of normal donors and in effusion T cells of the patients. After enrichment of autorosette-forming cells, there was no difference in AMLR of normal blood and cancer blood and effusions. These results indicate that the loss of AMLR in the blood of cancer patients is due to a reduction of number of autoreactive T cells and not to a defect of autologous stimulator non-T cells.


Higher early mortality with simultaneous rather than staged bilateral TKAs: results from the Swedish Knee Arthroplasty Register.
Patients with knee osteoarthritis (OA) often present with symptoms that warrant bilateral TKAs. There are potential benefits to operating on both knees on the same day, but the safety of simultaneous bilateral TKAs has been questioned. To evaluate whether there were any differences in 30-day mortality between patients having simultaneous bilateral TKAs and those having staged bilateral TKAs, we analyzed data from the Swedish Knee Arthroplasty Register and the Swedish Cause of Death Register. We included 48,931 patients with OA having 60,062 primary TKAs during 1985 to 2004; 1139 had surgery on both knees on the same day (simultaneous bilateral) and 3432 had surgery on both knees on two different occasions with less than 1 year between operations (staged bilateral). The 30-day mortality after simultaneous bilateral TKAs was 7.53 (confidence interval, 2.62-21.69) times higher than after the second of staged TKA and 3.77 (confidence interval, 2.04-6.98) times higher than after a primary unilateral TKA. Assuming the total risk for a staged procedure is twice that of a unilateral procedure, the risk of mortality within 30 days is 1.94 (confidence interval, 1.05-3.59) times higher with simultaneous than staged TKA. It is safer to operate on one knee at a time. Level II, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.


Sporadic Fatal Insomnia in Europe: Phenotypic Features and Diagnostic Challenges.
Comprehensively describe the phenotypic spectrum of sporadic fatal insomnia (sFI) to facilitate diagnosis and management of this rare and peculiar prion disorder. A survey among major prion disease reference centers in Europe identified 13 patients diagnosed with sFI in the past 20 years. We undertook a detailed analysis of clinical and histopathological features and the results of diagnostic investigations. Mean age at onset was 43 years, and mean disease duration 30 months. Early clinical findings included psychiatric, sleep, and oculomotor disturbances, followed by cognitive decline and postural instability. In all tested patients, video-polysomnography demonstrated a severe reduction of total sleep time and/or a disorganized sleep. Cerebrospinal fluid (CSF) levels of proteins 14-3-3 and t-tau were unrevealing, the concentration of neurofilament light protein (NfL) was more consistently increased, and the real-time quaking-induced conversion assay (RT-QuIC) revealed a positive prion seeding activity in 60% of cases. Electroencephalography and magnetic resonance imaging showed nonspecific findings, whereas fluorodeoxyglucose positron emission tomography (FDG-PET) demonstrated a profound bilateral thalamic hypometabolism in 71% of cases. Molecular analyses revealed PrPSc type 2 and methionine homozygosity at PRNP codon 129 in all cases. sFI is a disease of young or middle-aged adults, which is difficult to reconcile with the hypothesis of a spontaneous etiology related to stochastic, age-related PrP misfolding. The combination of psychiatric and/or sleep-related symptoms with oculomotor abnormalities represents an early peculiar clinical feature of sFI to be valued in the differential diagnosis. Video-polysomnography, FDG-PET, and especially CSF prion RT-QuIC and NfL constitute the most promising supportive diagnostic tests in vivo. Ann Neurol 2018;84:347-360.


Robot-assisted resection of gastric duplication cysts in a child: a detailed case report.
Gastric duplication cysts (GDCs) represent 4-9% of alimentary tract duplications. Early diagnosis and surgical excision are essential to avoid morbidity or neoplastic degeneration. Roboticassisted excision of GDCs has never been described in childhood. We report an asymptomatic male patient with 2 gastric cystic masses at ultrasonography (US)-study (diameter 25mm and 8mm), increasing in size at follow-up. At 20 months of age, magnetic- resonance-imaging-scan confirmed 2 round gastric masses (44×35mm and 16×12mm, respectively). Two months later, an elective robotic-assisted excision of GDCs was completed without complications. The patient was discharged at day 6 after procedure. Histology confirmed the diagnosis of GDCs. At a 2-year follow- up, US-study did not evidence any issue. In this first reported case of robotic-assisted cystectomy for CGD in childhood, the procedure seems safe, effective, and feasible. This approach improves the movements of the surgical instruments with better 3- D visualization in comparison with the laparoscopic approach.


Identification of a Kdn biosynthesis pathway in the haptophyte Prymnesium parvum suggests widespread sialic acid biosynthesis among microalgae.
Sialic acids are a family of more than 50 structurally distinct acidic sugars on the surface of all vertebrate cells where they terminate glycan chains and are exposed to many interactions with the surrounding environment. In particular, sialic acids play important roles in cell-cell and host-pathogen interactions. The sialic acids or related nonulosonic acids have been observed in Deuterostome lineages, Eubacteria, and Archaea but are notably absent from plants. However, the structurally related C8 acidic sugar 3-deoxy-d-manno-2-octulosonic acid (Kdo) is present in Gram-negative bacteria and plants as a component of bacterial lipopolysaccharide and pectic rhamnogalacturonan II in the plant cell wall. Until recently, sialic acids were not thought to occur in algae, but as in plants, Kdo has been observed in algae. Here, we report the de novo biosynthesis of the deaminated sialic acid, 3-deoxy-d-glycero-d-galacto-2-nonulosonic acid (Kdn), in the toxin-producing microalga Prymnesium parvum Using biochemical methods, we show that this alga contains CMP-Kdn and identified and recombinantly expressed the P. parvum genes encoding Kdn-9-P synthetase and CMP-Kdn synthetase enzymes that convert mannose-6-P to CMP-Kdn. Bioinformatics analysis revealed sequences related to those of the two P. parvum enzymes, suggesting that sialic acid biosynthesis is likely more widespread among microalgae than previously thought and that this acidic sugar may play a role in host-pathogen interactions involving microalgae. Our findings provide evidence that P. parvum has the biosynthetic machinery for de novo production of the deaminated sialic acid Kdn and that sialic acid biosynthesis may be common among microalgae.


Genetic determinants of autism in individuals with deletions of 18q.
Previous research has suggested that individuals with constitutional hemizygosity of 18q have a higher risk of autistic-like behaviors. We sought to identify genomic factors located on chromosome 18 as well as other loci that correlate with autistic behaviors. One hundred and five individuals with 18q- were assessed by high-resolution oligo aCGH and by parental ratings of behavior on the Gilliam Autism Rating Scale. Forty-five individuals (43%) had scores within the "possibly" or "very likely" categories of risk for an autism diagnosis. We searched for genetic determinants of autism by (1) identifying additional chromosome copy number changes (2) Identifying common regions of hemizygosity on 18q, and (3) evaluating four regions containing candidate genes located on 18q (MBD1, TCF4, NETO1, FBXO15). Three individuals with a "very likely" probability of autism had a captured 17p telomere in addition to the 18q deletion suggesting a possible synergy between hemizygosity of 18q and trigosity of 17p. In addition, two of the individuals with an 18q deletion and a "very likely" probability of autism rating had a duplication of the entire short arm of chromosome 18. Although no common region of hemizygosity on 18q was identified, analysis of four regions containing candidate genes suggested that individuals were significantly more likely to exhibit autistic-like behaviors if their region of hemizygosity included TCF4, NETO1, and FBXO15 than if they had any other combination of hemizygosity of the candidate genes. Taken together, these findings identify several new potential candidate genes or regions for autistic behaviors.


Facile synthesis of cellulose microfibers supported palladium nanospindles on graphene oxide for selective detection of dopamine in pharmaceutical and biological samples.
The cost-effective synthesis of novel functional nanomaterials has received significant attention in the physical and chemical sciences due to their improved surface area, high catalytic activity along with unique morphological features. This paper reports a facile and eco-friendly synthesis of spindle-like palladium nanostructures (PdSPs) on graphene oxide-cellulose microfiber (GO-CMF) composite for the first time. The GO-CMF/PdSPs composite was synthesized by an electrochemical method without the use of additional surfactants and capping agents. The synthesized materials were characterized and confirmed by using transmission electron microscopy, high-resolution scanning electron microscopy, X-ray diffraction spectroscopy, Raman spectroscopy and Fourier-transform infrared spectroscopy. As-synthesized GO-CMF/PdSPs composite modified electrode was used as a selective electrocatalyst for the oxidation of dopamine (DA). The electrochemical redox behaviors of DA were investigated using cyclic voltammetry (CV). The CV results revealed that the GO-CMF/PdSPs composite modified electrode has 10 folds enhanced oxidation current response to DA than GO, PdSPs and GO-CMF modified GCEs. Under optimized conditions, the GO-CMF/PdSPs composite sensor exhibits a linear response to DA in the concentration range from 0.3 to 196.3 μM with the lower detection limit of 23 nM. The nanocomposite electrode also shows promising features towards the reliable and selective detection of DA, which includes high stability, reproducibility and high selectivity towards the commonly interfering species such as ascorbic acid, uric acid, and dihydroxybenzene isomers. The sensor was successfully tested for the real-time detection of DA in the commercial DA injections and human serum samples.


Celastrol inhibits colon cancer cell proliferation by downregulating miR-21 and PI3K/AKT/GSK-3β pathway.
Celastrol is a traditional Chinese medicine, that is derived from Tripterygium wilfordii. It has been proposed to offer anti-tumor potential. MicroRNAs also play important roles in tumorigenesis. However, the anti-tumor mechanism of Celastrol and whether miRNAs are involved in the process are still unknown. In the present study, MTT assay was used to test the IC50 of Celastrol and cell viability. PCNA, PI3K, Akt, GSK3β, phosphorylated Akt and GSK3β were measured by western blotting. Flow cytometry was introduced to detect the apoptosis. We found Celastrol inhibited colon cancer cell viability in a dose-dependent manner companied with PCNA downregulation. Apoptosis was induced by Celastrol. After Celastrol treatment, BCL-2 expression decreased while BAX increased and the Caspase-3 activity was induced. Simultaneously, miR-21 expression was reduced in Celastrol-treated colon cancer cells. miR-21 mimic overexpression could enhance the cell viability, inhibit the apoptosis, decrease BCL-2 expression, increase BAX and induce Caspase-3 activity to some extent which were reversed by Celastrol. In addition, the PI3K/AKT/GSK-3β pathway was activated by miR-21 mimic but partially arrested by extra-adding Celastrol. Thus, Celastrol may inhibit colon cancer cell proliferation by negatively regulating miR-21 and the PI3K/AKT/GSK-3β pathway.


TgMORN1 is a key organizer for the basal complex of Toxoplasma gondii.
Toxoplasma gondii is a leading cause of congenital birth defects, as well as a cause for ocular and neurological diseases in humans. Its cytoskeleton is essential for parasite replication and invasion and contains many unique structures that are potential drug targets. Therefore, the biogenesis of the cytoskeletal structure of T. gondii is not only important for its pathogenesis, but also of interest to cell biology in general. Previously, we and others identified a new T. gondii cytoskeletal protein, TgMORN1, which is recruited to the basal complex at the very beginning of daughter formation. However, its function remained largely unknown. In this study, we generated a knock-out mutant of TgMORN1 (DeltaTgMORN1) using a Cre-LoxP based approach. We found that the structure of the basal complex was grossly affected in DeltaTgMORN1 parasites, which also displayed defects in cytokinesis. Moreover, DeltaTgMORN1 parasites showed significant growth impairment in vitro, and this translated into greatly attenuated virulence in mice. Therefore, our results demonstrate that TgMORN1 is required for maintaining the structural integrity of the parasite posterior end, and provide direct evidence that cytoskeleton integrity is essential for parasite virulence and pathogenesis.


[Effectiveness of antimicrobial formulations for acne based on orange (Citrus sinensis) and sweet basil (Ocimum basilicum L) essential oils].
Currently, the antimicrobial resistance has developed in bacterial strains involved in the development of acne. Therefore, alternatives to antibiotic treatment have become necessary. Gel formulations were designed based on essential oils and acetic acid, and their effectiveness was evaluated in patients affected by acne. Masked simple experimental study of three gel formulations on 28 volunteer patients, separated in four groups of seven patients. Treatments were applied daily for eight weeks and consisted of (1) antibacterial (essential oils), (2) keratolytic medication (3) essential oils mixed with acetic acetic, and (4) kerolytic medication with acetic acid. Weekly checks were conducted to evaluate patient improvement. All groups reported an improvement of the acne condition, which ranged between 43% and 75% clearance of lesions. Evidence of treatment disappeared within minutes, showing little discomfort or side effects after application. The essential oil formulations were chemically and physically stable during application of treatments. This was demonstrated by gas chromatography, where no evidence no change neither the composition profiles of essential oils nor in acetic acid. The results were ranked good to excellent, particularly for the acetic acid mixture, which achieved improvements of 75%. This appeared to be a result of their joint antiseptic and keratolytic activity. Side effects (burning and redness) disappeared within a few minutes of completing the application, therefore, did not interfere with adherence to treatment.


Immune response to recombinant Escherichia coli Iss protein in poultry.
Colibacillosis accounts for significant losses to the poultry industry, and control efforts are hampered by limited understanding of the mechanisms used by avian pathogenic Escherichia coli (APEC) to cause disease. We have found that the presence of the increased serum survival gene (iss) is strongly associated with APEC but not with commensal E. coli, making iss, and the protein it encodes (Iss), candidate targets of colibacillosis control procedures. To assess the potential of Iss to elicit a protective response in chickens against APEC challenge, Iss fusion proteins were produced and administered subcutaneously to four groups of 2-wk-old specific-pathogen-free leghorn chickens. At 4 wk postimmunization, birds were challenged with APEC from serogroups 02 and 078 via intramuscular injection. At 2 wk postchallenge, birds were necropsied, and lesions consistent with colibacillosis were scored. Also, sera were collected from the birds pre- and postimmunization, and antibody titers to Iss were determined. Immunized birds produced a humoral response to Iss, and they had significantly lower lesion scores than the unimmunized control birds following challenge with both APEC strains. Birds that received the smallest amount of immunogen had the lowest lesion scores. Although further study will be needed to confirm the value of Iss as an immunoprotective antigen, these preliminary data suggest that Iss may have the potential to elicit significant protection in birds against heterologous E. coli challenge.


A comparison of the antimicrobial activity and toxicity of six combretum and two terminalia species from southern Africa.
Plants of the family Combretaceae are amongst the most widely used plants for traditional medicinal purposes in southern Africa. In particular, many species of Combretum and Terminalia are used for their antibacterial, antifungal, antiprotozoal, antiviral, antidiarrhoeal, analgesic, antimalarial, antioxidant, anti-inflammatory and anticancer activities, yet their antimicrobial potential has not been rigorously studied and compared. A survey of antimicrobial activity was undertaken on selected South African Combretum and Terminalia species. Sixteen extracts from 6 Combretum and 2 Terminalia plant species with a history of medicinal usage were investigated by disc diffusion assay against a panel of bacteria and fungi and their MIC values were determined. Toxicity was determined using the Artemia franciscana nauplii bioassay. All extracts tested displayed broad spectrum antibacterial activity, inhibiting the growth of 12-16 (75-100%) of the bacteria tested, with Gram-positive and Gram-negative bacteria being approximately equally susceptible. Potent antibacterial activities (generally in the range 200-5000 μg/ml) were evident for all Combretaceae extracts against both Gram-positive and Gram-negative bacteria. Similarly, the extracts also displayed good antifungal activity, inhibiting the growth of 2-3 (66.7-100%) of the fungal species tested, with fungal growth inhibition activities generally in the range 200-4000 μg/ml. In general, the Terminalia extracts had better efficacies than the Combretum extracts. Furthermore, the methanol extracts were generally better antimicrobial agents than the water extracts. All extracts were also shown to be non-toxic in the Artemia nauplii bioassay. The lack of toxicity of these extracts and their inhibitory bioactivity against a panel of bacteria and fungi indicate their potential as medicinal agents and partially validate their usage in multiple South African traditional medicinal systems.


Measuring time utilization of pharmacists in the Birmingham Free Clinic dispensary.
Free and charitable clinics are a critical part of America's healthcare safety net. Although informatics tools have the potential to mitigate many of the organizational and service-related challenges facing these clinics, little research attention has been paid to the workflows and potential impact of electronic systems in these settings. In previous work, we performed a qualitative investigation at a free clinic dispensary to identify workflow challenges that may be alleviated through introduction of informatics interventions. However, this earlier study did not quantify the magnitude of these challenges. Time-motion studies offer a precise standard in quantifying healthcare workers' time expenditures on clinical activities, and can provide valuable insight into system specifications. These data, informed by a lean healthcare perspective, provide a quality improvement framework intended to maximize value and eliminate waste in inefficient workflow processes. We performed a continuous observation time-motion study in the Birmingham Free Clinic dispensary. Two researchers followed pharmacists over the course of three general clinic sessions and recorded the duration of specific tasks. Pharmacists were then asked to identify tasks as value-added or non-value-added to facilitate calculation of the value quotient, a metric used to determine a workflow's level of efficiency. Four high-level workflow categories occupied almost 95 % of pharmacist time: prescription (Rx) preparation (39.8 %), clinician interaction (21.5 %), EMR operations (14.8 %), and patient interaction (18.7 %). Pharmacists invested the largest portion of time in prescription preparation, with 21.8 % of pharmacist time spent handwriting medication labels. Based on value categorizations made by the pharmacists, the average value quotient was found to be 40.3 %, indicating that pharmacists spend more than half of their time completing tasks they consider to be non-value-added. Our results show that pharmacists spend a large portion of their time preparing prescriptions, primarily the handwritten labeling of medication bottles and documentation tasks, which is not an optimal utilization of pharmacist expertise. The value quotient further supports that there are many wasteful tasks that may benefit from workflow redesign and health information technology, which could result in efficiency improvements for pharmacists.


Mycobacterium tuberculosis infection in psoriatic patients treated with biologics: Real-world data from 18 Japanese facilities.
Although rare, tuberculosis has been reported with biologic treatment against psoriasis in Japan, a tuberculosis medium-burden country. Mycobacterial infection often develops after a long incubation period and might not have been adequately identified in clinical trials or post-marketing surveillance. To determine the real-world incidence of tuberculosis in psoriatic patients treated with biologics, we conducted a retrospective, multicenter, observational study in 18 facilities in Western Japan. Psoriatic patients who visited a participating facility between 2010 and March 2017 and received biologic reagents were enrolled. Information on sex, age at first biologic treatment, results of interferon-γ release assay (IGRA) for Mycobacterium tuberculosis, treatment history with isoniazid, and onset of active and/or latent tuberculosis was collected. A total of 1117 patients (830 men and 287 women) were enrolled. The mean duration of biologic treatment was 3.54 years. Sixty-five patients (5.8%) showed positive IGRA results at screening. Active tuberculosis developed in two patients after the administration of tumor necrosis factor inhibitors (both involved miliary tuberculosis). Latent tuberculosis was observed in two patients treated with anti-interleukin-12/23p40 antibody. The incidence rate of tuberculosis, including latent tuberculosis, in this survey was 0.36%. Although the incidence rate of tuberculosis was low considering the observation period of biologic treatment, active tuberculosis was found in both the screening-negative group and a screening-positive subject after isoniazid prophylaxis (both miliary tuberculosis), concluding that negative screening or isoniazid treatment does not always assure that an individual has no tuberculosis. Hence, dermatologists still need to pay careful attention to tuberculosis at every patient visit.


Can training bar staff in responsible serving practices reduce alcohol-related harm?
A responsible service training programme aimed at reducing alcohol-related harm was implemented in a popular entertainment area over several months in 1992-93. Another popular entertainment area provided a control site. A number of evaluation measures were used: breath tests on 872 patrons from selected venues; drink driving data; risk assessments; the use of 'pseudo patrons'; and knowledge and attitude changes among trained bar staff (n = 88). Compared to control sites the intervention sites showed an immediate pre- to post-test reduction in patrons rated by researchers as extremely drunk and an eventual reduction from pre-test to follow-up in patrons with blood alcohol levels > = 0.08. There was also a small but significant increase in knowledge among bar staff. There was no significant reduction in patrons with blood alcohol levels > = 0.15 or in the number of drink driving offences from intervention sites during the study period. Pseudo drunk patrons were rarely refused service, identification was rarely checked and non-photographic identification was accepted on most occasions. The less than satisfactory outcome is attributed to poor implementation of the training and a lack of support among managers. The positive results from one venue, whose manager embraced the programme, served to highlight the importance of management support. It is suggested that mandatory training and routine enforcement of licensing laws are essential if the goals of responsible serving are to be met.


An overview of the effects of anti-IgE therapies.
Omalizumab, a humanized mAb that binds to the CH3 domain near the binding site for the high-affinity type-I IgE Fc receptors of human IgE, can neutralize free IgE and inhibit the IgE allergic pathway without sensitizing mast cells and basophils. We found that omalizumab in patients with severe persistent asthma (SPA) was an effective therapy for asthma and the following co-morbid conditions: chronic urticaria (CU), bee venom allergy, latex allergy, atopic dermatitis, food allergy and Samter's syndrome. Information on the use of omalizumab in treatment of asthma and other allergic diseases has improved our understanding that treatment acts on many levels, including regulating levels of inflammatory proteins, including cytokines (copper-containing alpha- 2-glycoprotein, total antioxidant capacity, MDA, NO, H2O2, CXCL8, IL-10, TGF-β, GMCSF, IL-17, IL-1β), MPV, Hs-CRP, eosinophil cationic peptide, vitamin-D (25(OH)D), homocysteine (Hcy), OX-2, d- dimer, albumin, and sApo-2L. The decrease in Hcy concentrations and increase in 25(OH)D also support the existence of a vascular endothelial protection mechanism. Mediators and cells classically involved in pro-coagulant and anticoagulant pathways together play a role in SPA and CU pathophysiology and omalizumab effect. The mechanism of action of omalizumab in the treatment of asthma is believed to be multifactorial, and includes effects mediated through altered production of redox metabolites, extrinsic coagulation pathway, oxidative markers-related mi RNA, TRAIL-related mi RNA, and regulation of production of known inflammatory proteins.


Hypocholesterolemic effect of hot-water extract from mycelia of Cordyceps sinensis.
This study was conducted to investigate the hypocholesterolemic effect of the hot-water fraction (HW) from cultured mycelia of Cordyceps sinensis in a 5 l fermenter. The composition of HW was mainly carbohydrate (83.9%) and protein (11.8%) on a dry basis, and the carbohydrate of HW consisted of glucose, mannose, galactose, and arabinose in the molecular ratio of 1.0 : 0.8 : 0.5 : 0.1, respectively. In mice fed a cholesterol-free diet and those fed a cholesterol-enriched diet, body and liver weights were not significantly different from those of the controls. The serum total cholesterol (TC) of all mice groups administered HW (150 and 300 mg/kg/d, respectively) with the cholesterol-enriched diet decreased more than in the control group. Among the mice fed the cholesterol-enriched diet, HW also increased the high-density lipoprotein (HDL) cholesterol level, but decreased the very low-density lipoprotein plus low-density lipoprotein (VLDL+LDL) cholesterol level. The changes in HDL- and VLDL+LDL-cholesterol levels consequently decreased the atherogenic value. The results indicate that HW in rats administered a cholesterol-enriched diet decreased the plasma cholesterol level. The 300 mg/kg dose had a significant effect on the serum TC level.


Association between higher plasma lutein, zeaxanthin, and vitamin C concentrations and longer telomere length: results of the Austrian Stroke Prevention Study.
To examine the association between plasma concentrations of antioxidative micronutrients and leukocyte telomere length (LTL) in elderly adults. Cross-sectional cohort study. Austrian Stroke Prevention Study, a population-based cohort study on brain aging. Individuals with a mean age of 66 ± 7 (n = 786; 58% female). Concentrations of vitamin C, lutein, zeaxanthin, β-cryptoxanthin, canthaxanthin, lycopene, α- and γ-tocopherol, α- and β-carotene, and retinol in plasma, advanced oxidation protein products as a measure of oxidative stress in serum, and LTL were measured. Vitamins and carotenoids were measured using high-performance liquid chromatography, advanced oxidation protein products using spectrophotometry, and telomere length using quantitative real-time polymerase chain reaction. Multiple linear regression analyses with adjustment for age and sex demonstrated that higher lutein, zeaxanthin, and vitamin C concentrations were strongly associated with longer telomere length. The associations were independent of body mass index, maximum oxygen uptake, and vascular risk factors and were not mediated by advanced oxidation protein products content. This study provides first evidence that higher lutein, zeaxanthin, and vitamin C concentrations in plasma are associated with longer LTL in normal elderly persons and suggest a protective role of these vitamins in telomere maintenance.


Coding of position by simultaneously recorded sensory neurones in the cat dorsal root ganglion.
Muscle, cutaneous and joint afferents continuously signal information about the position and movement of individual joints. How does the nervous system extract more global information, for example about the position of the foot in space? To study this question we used microelectrode arrays to record impulses simultaneously from up to 100 discriminable nerve cells in the L6 and L7 dorsal root ganglia (DRG) of the anaesthetized cat. When the hindlimb was displaced passively with a random trajectory, the firing rate of the neurones could be predicted from a linear sum of positions and velocities in Cartesian (x, y), polar or joint angular coordinates. The process could also be reversed to predict the kinematics of the limb from the firing rates of the neurones with an accuracy of 1-2 cm. Predictions of position and velocity could be combined to give an improved fit to limb position. Decoders trained using random movements successfully predicted cyclic movements and movements in which the limb was displaced from a central point to various positions in the periphery. A small number of highly informative neurones (6-8) could account for over 80% of the variance in position and a similar result was obtained in a realistic limb model. In conclusion, this work illustrates how populations of sensory receptors may encode a sense of limb position and how the firing of even a small number of neurones can be used to decode the position of the limb in space.


A simple protocol for platelet-mediated clumping of Plasmodium falciparum-infected erythrocytes in a resource poor setting.
P. falciparum causes the majority of severe malarial infections. The pathophysiological mechanisms underlying cerebral malaria (CM) are not fully understood and several hypotheses have been put forward, including mechanical obstruction of microvessels by P. falciparum-parasitized red blood cells (pRBC). Indeed, during the intra-erythrocytic stage of its life cycle, P. falciparum has the unique ability to modify the surface of the infected erythrocyte by exporting surface antigens with varying adhesive properties onto the RBC membrane. This allows the sequestration of pRBC in multiple tissues and organs by adhesion to endothelial cells lining the microvasculature of post-capillary venules (1). By doing so, the mature forms of the parasite avoid splenic clearance of the deformed infected erythrocytes (2) and restrict their environment to a more favorable low oxygen pressure (3). As a consequence of this sequestration, it is only immature asexual parasites and gametocytes that can be detected in peripheral blood. Cytoadherence and sequestration of mature pRBC to the numerous host receptors expressed on microvascular beds occurs in severe and uncomplicated disease. However, several lines of evidence suggest that only specific adhesive phenotypes are likely to be associated with severe pathological outcomes of malaria. One example of such specific host-parasite interactions has been demonstrated in vitro, where the ability of intercellular adhesion molecule-1 to support binding of pRBC with particular adhesive properties has been linked to development of cerebral malaria (4,5). The placenta has also been recognized as a site of preferential pRBC accumulation in malaria-infected pregnant women, with chondrotin sulphate A expressed on syncytiotrophoblasts that line the placental intervillous space as the main receptor (6). Rosetting of pRBC to uninfected erythrocytes via the complement receptor 1 (CD35)(7,8) has also been associated with severe disease (9). One of the most recently described P. falciparum cytoadherence phenotypes is the ability of the pRBC to form platelet-mediated clumps in vitro. The formation of such pRBC clumps requires CD36, a glycoprotein expressed on the surface of platelets. Another human receptor, gC1qR/HABP1/p32, expressed on diverse cell types including endothelial cells and platelets, has also been shown to facilitate pRBC adhesion on platelets to form clumps (10). Whether clumping occurs in vivo remains unclear, but it may account for the significant accumulation of platelets described in brain microvasculature of Malawian children who died from CM (11). In addition, the ability of clinical isolate cultures to clump in vitro was directly linked to the severity of disease in Malawian (12) and Mozambican patients (13), (although not in Malian (14)). With several aspects of the pRBC clumping phenotype poorly characterized, current studies on this subject have not followed a standardized procedure. This is an important issue because of the known high variability inherent in the assay (15). Here, we present a method for in vitro platelet-mediated clumping of P. falciparum with hopes that it will provide a platform for a consistent method for other groups and raise awareness of the limitations in investigating this phenotype in future studies. Being based in Malawi, we provide a protocol specifically designed for a limited resource setting, with the advantage that freshly collected clinical isolates can be examined for phenotype without need for cryopreservation.


Increased immunogenicity of a DNA-launched Venezuelan equine encephalitis virus-based replicon DNA vaccine.
A novel genetic vaccine that is based on a Venezuelan equine encephalitis virus (VEE) replicon launched from plasmid DNA is described. The plasmid encodes a VEE replicon under the transcriptional control of the cytomegalovirus immediate-early promoter (VEE DNA). The VEE DNA consistently expressed 3- to 15-fold more green fluorescent protein in vitro than did a conventional DNA vaccine. Furthermore, transfection with the DNA-launched VEE replicon induced apoptosis and type I interferon production. Inoculation of mice with VEE DNA encoding human immunodeficiency virus type 1 gp160 significantly increased humoral responses by several orders of magnitude compared to an equal dose of a conventional DNA vaccine. These increases were also observed at 10- and 100-fold-lower doses of the VEE DNA. Cellular immune responses measured by gamma interferon and interleukin 2 enzyme-linked immunospot assay were significantly higher in mice immunized with the VEE DNA at decreased doses. The immune responses induced by the VEE DNA-encoded antigen, however, were independent of an intact type I interferon signaling pathway. Moreover, the DNA-launched VEE replicon induced an efficient prime to a VEE replicon particle (VRP) boost, increasing humoral and cellular immunity by at least 1 order of magnitude compared to VEE DNA only. Importantly, immunization with VEE DNA, as opposed to VRP, did not induce any anti-VRP neutralizing antibodies. Increased potency of DNA vaccines and reduced vector immunity may ultimately have an impact on the design of vaccination strategies in humans.


Bone-implant contact at calcium phosphate-coated and porous titanium oxide (TiUnite)-modified oral implants.
Calcium phosphate (CP)-coated implants are usually referred to as having osteoconductive properties, whereas titanium implants with a native oxide layer are considered less osteoconductive. Often smooth titanium oxides (TOs) are compared to relatively rough CP structures. The objective of this study was to evaluate osteoconduction by comparing bone-implant contact at a relatively smooth, highly crystalline CP coating with a structured, porous TO (TiUnite)-modified surface. Ten adult Hound Labrador mongrel dogs were used. Four titanium implants (Nobel Biocare) with CP-coated (2) or TO-modified (2) surfaces were installed 12 weeks following mandibular premolar and molar teeth extraction. The implants were alternated within and between jaw quadrants in consecutive animals. Mucosal flaps were advanced and sutured leaving the implants in a submerged position. The animals were injected with fluorescent bone labels at 3 and 4 weeks postsurgery, and pre-euthanasia to monitor progress of bone formation. The animals were euthanized at 8 weeks postsurgery and block biopsies were prepared for histologic and histometric analysis. There were no remarkable differences in bone formation and apparent bone-implant contact comparing the TO-modified and CP-coated surfaces. However, the measured average bone-implant contact was 71% and 57% (P=0.027) for TO-modified and CP-coated implants, respectively. We conclude that the TO surface exhibits osteoconductive properties exceeding that of the CP surface. One or several of the chemical and physical properties of the TO surface may result in the remarkable bone formation along its surface. This study indicated that crystallinity and/or chemistry may be important.


Ross, macdonald, and a theory for the dynamics and control of mosquito-transmitted pathogens.
Ronald Ross and George Macdonald are credited with developing a mathematical model of mosquito-borne pathogen transmission. A systematic historical review suggests that several mathematicians and scientists contributed to development of the Ross-Macdonald model over a period of 70 years. Ross developed two different mathematical models, Macdonald a third, and various "Ross-Macdonald" mathematical models exist. Ross-Macdonald models are best defined by a consensus set of assumptions. The mathematical model is just one part of a theory for the dynamics and control of mosquito-transmitted pathogens that also includes epidemiological and entomological concepts and metrics for measuring transmission. All the basic elements of the theory had fallen into place by the end of the Global Malaria Eradication Programme (GMEP, 1955-1969) with the concept of vectorial capacity, methods for measuring key components of transmission by mosquitoes, and a quantitative theory of vector control. The Ross-Macdonald theory has since played a central role in development of research on mosquito-borne pathogen transmission and the development of strategies for mosquito-borne disease prevention.


Host factors and failure of interferon-alpha treatment in hepatitis C virus.
Failure of interferon-alpha (IFN-alpha) treatment in patients with chronic hepatitis C virus (HCV) infection is a challenging obstacle for clinical and experimental hepatology. Both viral and host factors have been implicated in reducing responsiveness to IFN-alpha therapy. The role of viral factors has been studied extensively and has been summarized in several review articles; however, much less attention has been paid to host factors. In this paper, we review evidence of host factor involvement in IFN-alpha treatment failure. We discuss possible underlying mechanisms responsible for these effects. Potential therapeutic strategies to enhance the effectiveness of IFN-alpha therapy for HCV are also proposed.


Rational considerations about development of live attenuated Yersinia pestis vaccines.
The risk of plague as a bioweapon has prompted increasing research efforts to develop plague vaccines due to its extreme virulence and the ease of its transmission. Subunit vaccines that contain F1 and LcrV antigens of Y. pestis have been tested for safety and immunogenicity, but doubts have been raised about whether subunit vaccines that engender antibody responses will protect against pneumonic plague, which requires both humeral and cellular immune responses for protection. The live, attenuated vaccine EV76, a pgm locus deficient Y. pestis strain, has been used for a long time in the Former Soviet Union and some Asian countries, but is not commercially available in the US and Europe due to safety concerns. However, the live attenuated Y. pestis vaccines are still considered to be the most effective way to prevent plague. In this review, we present our opinions about rationally creating live, safe and immunogenic Y. pestis vaccines with potential use for human based on established researches.


A CCaMK-CYCLOPS-DELLA Complex Activates Transcription of RAM1 to Regulate Arbuscule Branching.
Intracellular arbuscular mycorrhiza symbiosis between plants and glomeromycotan fungi leads to formation of highly branched fungal arbuscules that release mineral nutrients to the plant host. Their development is regulated in plants by a mechanistically unresolved interplay between symbiosis, nutrient, and hormone (gibberellin) signaling. Using a positional cloning strategy and a retrotransposon insertion line, we identify two novel alleles of Lotus japonicus REDUCED ARBUSCULAR MYCORRHIZA1 (RAM1) encoding a GRAS protein. We confirm that RAM1 is a central regulator of arbuscule development: arbuscule branching is arrested in L. japonicus ram1 mutants, and ectopic expression of RAM1 activates genes critical for arbuscule development in the absence of fungal symbionts. Epistasis analysis places RAM1 downstream of CCaMK, CYCLOPS, and DELLA because ectopic expression of RAM1 restores arbuscule formation in cyclops mutants and in the presence of suppressive gibberellin. The corresponding proteins form a complex that activates RAM1 expression via binding of CYCLOPS to a cis element in the RAM1 promoter. We thus reveal a transcriptional cascade in arbuscule development that employs the promoter of RAM1 as integrator of symbiotic (transmitted via CCaMK and CYCLOPS) and hormonal (gibberellin) signals.


Antidepressant stimulation of CDP-diacylglycerol synthesis does not require monoamine reuptake inhibition.
Recent studies demonstrate that diverse antidepressant agents increase the cellular production of the nucleolipid CDP-diacylglycerol and its synthetic derivative, phosphatidylinositol, in depression-relevant brain regions. Pharmacological blockade of downstream phosphatidylinositide signaling disrupted the behavioral antidepressant effects in rats. However, the nucleolipid responses were resistant to inhibition by serotonin receptor antagonists, even though antidepressant-facilitated inositol phosphate accumulation was blocked. Could the neurochemical effects be additional to the known effects of the drugs on monoamine transmitter transporters? To examine this question, we tested selected agents in serotonin-depleted brain tissues, in PC12 cells devoid of serotonin transporters, and on the enzymatic activity of brain CDP-diacylglycerol synthase - the enzyme that catalyzes the physiological synthesis of CDP-diacylglycerol. Imipramine, paroxetine, and maprotiline concentration-dependently increased the levels of CDP-diacylglycerol and phosphatidylinositides in PC12 cells. Rat forebrain tissues depleted of serotonin by pretreatment with p-chlorophenylalanine showed responses to imipramine or maprotiline that were comparable to respective responses from saline-injected controls. With fluoxetine, nucleolipid responses in the serotonin-depleted cortex or hippocampus were significantly reduced, but not abolished. Each drug significantly increased the enzymatic activity of CDP-diacylglycerol synthase following incubations with cortical or hippocampal brain tissues. Antidepressants probably induce the activity of CDP-diacylglycerol synthase leading to increased production of CDP-diacylglycerol and facilitation of downstream phosphatidylinositol synthesis. Phosphatidylinositol-dependent signaling cascades exert diverse salutary effects in neural cells, including facilitation of BDNF signaling and neurogenesis. Hence, the present findings should strengthen the notion that modulation of brain phosphatidylinositide signaling probably contributes to the molecular mechanism of diverse antidepressant medications.


The effect of a novel viscous polysaccharide along with lifestyle changes on short-term weight loss and associated risk factors in overweight and obese adults: an observational retrospective clinical program analysis.
Viscous soluble dietary fiber has been demonstrated to reduce postprandial glycemia and may promote satiety. PolyGlycopleX (PGX) is a highly viscous polysaccharide manufactured by reacting glucomannan with other soluble polysaccharides using a proprietary process (EnviroSimplex). The resulting polysaccharide (alpha-D-glucurono-alpha-D-manno-beta-D-manno-beta-D-glucan, alpha-L-gulurono-beta-D-mannuronan, beta-D-gluco-beta-D-mannan, alpha-D-glucurono-alpha-D-manno-beta-D-manno-beta-D-gluco, alpha-L-gulurono-beta-D-mannurono, beta-D-gluco-beta-D-mannan) is a novel entity with the highest viscosity and water-holding capacity of currently known fibers. A total of 29 sedentary overweight or obese adults (23 women; six men), ages 20-65 with a body mass index (BMI) range of 25 kg/m(2) to 36 kg/m(2) participated in a clinical weight-loss program. PGX (5 g) was consumed with 500 mL water, 5-10 minutes before each meal, 2-3 times daily for 14 weeks. Significant reductions were observed (p less than 0.05) in weight (-5.79 +/- 3.55 kg), waist circumference (-12.07 +/- 5.56 cm), and percentage body fat (-2.43 +/- 2.39 percent) compared to baseline values. In addition, subjects employing PGX had a significant reduction of 19.26 percent (n=17; p less than 0.05) and 25.51 percent (n=16; p less than 0.05) in total and LDL plasma cholesterol values, respectively, at the end of the study period. The consumption of PGX in concert with lifestyle modifications may be a useful strategy for weight loss in overweight and obese individuals.


Cyclosporin A induces cardiac differentiation but inhibits hemato-endothelial differentiation of P19 cells.
Little is known about the mechanisms underlying the effects of Cyclosporin A (CsA) on the fate of stem cells, including cardiomyogenic differentiation. Therefore, we investigated the effects and the molecular mechanisms behind the actions of CsA on cell lineage determination of P19 cells. CsA induced cardiomyocyte-specific differentiation of P19 cells, with the highest efficiency at a concentration of 0.32 μM during embryoid body (EB) formation via activation of the Wnt signaling pathway molecules, Wnt3a, Wnt5a, and Wnt8a, and the cardiac mesoderm markers, Mixl1, Mesp1, and Mesp2. Interestingly, cotreatment of P19 cells with CsA plus dimethyl sulfoxide (DMSO) during EB formation significantly increases cardiac differentiation. In contrast, mRNA expression levels of hematopoietic and endothelial lineage markers, including Flk1 and Er71, were severely reduced in CsA-treated P19 cells. Furthermore, expression of Flk1 protein and the percentage of Flk1+ cells were severely reduced in 0.32 μM CsA-treated P19 cells compared to control cells. CsA significantly modulated mRNA expression levels of the cell cycle molecules, p53 and Cyclins D1, D2, and E2 in P19 cells during EB formation. Moreover, CsA significantly increased cell death and reduced cell number in P19 cells during EB formation. These results demonstrate that CsA induces cardiac differentiation but inhibits hemato-endothelial differentiation via activation of the Wnt signaling pathway, followed by modulation of cell lineage-determining genes in P19 cells during EB formation.


Beta-amyloid (beta/A4) deposition in the medial temporal lobe in Down's syndrome: effects of brain region and patient age.
The density of diffuse, primitive, classic and compact beta-amyloid (beta/A4) deposits was studied in the medial temporal lobe in 12 cases of Down's syndrome (DS) from 38 to 67 years of age. Total beta/A4 deposit density was greater in the adjacent cortex compared with regions of the hippocampus, and these differences were similar within each age group of patients. The ratio of the primitive to diffuse deposits was greater in the hippocampus than in the adjacent cortex. Total beta/A4 density did not vary significantly with patient age. However, the density of the diffuse deposits exhibited a parabolic, and the primitive, classic and compact deposits an inverted parabolic, response with age. Hence, in DS, (1) beta/A4 density remains relatively constant with age, (2) differences in beta/A4 density between the hippocampus and adjacent cortex are established at an early age, and (3) mature beta/A4 subtype formation depends on brain region and patient age.


Protecting Sentient Artificial Intelligence: A Survey of Lay Intuitions on Standing, Personhood, and General Legal Protection.
To what extent, if any, should the law protect sentient artificial intelligence (that is, AI that can feel pleasure or pain)? Here we surveyed United States adults (n = 1,061) on their views regarding granting 1) general legal protection, 2) legal personhood, and 3) standing to bring forth a lawsuit, with respect to sentient AI and eight other groups: humans in the jurisdiction, humans outside the jurisdiction, corporations, unions, non-human animals, the environment, humans living in the near future, and humans living in the far future. Roughly one-third of participants endorsed granting personhood and standing to sentient AI (assuming its existence) in at least some cases, the lowest of any group surveyed on, and rated the desired level of protection for sentient AI as lower than all groups other than corporations. We further investigated and observed political differences in responses; liberals were more likely to endorse legal protection and personhood for sentient AI than conservatives. Taken together, these results suggest that laypeople are not by-and-large in favor of granting legal protection to AI, and that the ordinary conception of legal status, similar to codified legal doctrine, is not based on a mere capacity to feel pleasure and pain. At the same time, the observed political differences suggest that previous literature regarding political differences in empathy and moral circle expansion apply to artificially intelligent systems and extend partially, though not entirely, to legal consideration, as well.


Aptabody-aptatope interactions in aptablotting assays.
We demonstrate an aptablotting assay method that involves direct and indirect aptabody recognition. Nanoscale single-stranded DNA aptamers against GST and DIG-tags are utilized as aptabodies (GST-2 and DIG-1, respectively), and the GST-2 aptabody binding site, or aptatope, as predicted by a MOE-docking simulation of the protein-aptamer complex, shows the interaction of the GST-2 aptabody at the catalytically active region. The aptabody-aptatope interaction was evaluated by an in vitro enzyme inhibitory analysis. The binding capacity of the GST-2 aptabody was assessed by dot-blot, EMSA and SDS-PAGE/electroblot analyses, and the results showed that the aptabodies interact with both the native mono-/dimeric form and the denatured GST form on a membrane. The use of aptabodies can overcome the obstacles of current immunoblot assays, and these molecules are easily assessable via ELISA systems. Moreover, the hybridization of aptabodies and antibodies (hybrid-aptablotting) may have considerable impacts on the design of bioassay platforms.


Comparative Analysis of Regions with Distorted Segregation in Three Diploid Populations of Potato.
Genes associated with gametic and zygotic selection could underlie segregation distortion, observed as alterations of expected Mendelian genotypic frequencies in mapping populations. We studied highly dense genetic maps based on single nucleotide polymorphisms to elucidate the genetic nature of distorted segregation in potato. Three intra- and interspecific diploid segregating populations were used. DRH and D84 are crosses between the sequenced doubled monoploid DM 1-3 516 R44 Solanum tuberosum Group Phureja and either RH89-039-16 S. tuberosum or 84SD22, a S. tuberosum × S. chacoense hybrid. MSX902 is an interspecific cross between 84SD22 and Ber83 S. berthaultii × 2 × species mosaic. At the 0.05 significance level, 21%, 57%, and 51% of the total markers mapped in DRH, D84, and MSX902 exhibited distorted segregation, respectively. Segregation distortion regions for DRH were located on chromosomes 9 and 12; for D84 on chromosomes 2, 3, 4, 6, 7, and 8; and on chromosomes 1, 2, 7, 9, and 12 for MSX902. In general, each population had unique segregation distortion regions and directions of distortion. Interspecific crosses showed greater levels of distorted segregation and lower recombination rates as determined from the male parents. The different genomic regions where the segregation distortion regions occurred in the three populations likely reflect unique genetic combinations producing distorted segregation.