Physical disability and social interaction: factors associated with low social contact and home confinement in disabled older women (The Women's Health and Aging Study).
This study examines the association of disability and social interaction, measured as in-person contact with non-household members and home confinement, and identifies sociodemographic, socioeconomic, and health-related factors that modify this relationship. Participants were 1,002 moderately to severely disabled community-dwelling women aged 65 and older from the Women's Health and Aging Study, identified by screening an age-stratified random sample of Medicare beneficiaries in Baltimore, Maryland. Logistic regression models were used to estimate the odds of low social interaction associated with disability and each independent modifier. In a typical week, 23% did not visit with anyone residing outside their households and 17% did not leave their homes. In addition to and independent of disability level, older age, not completing high school, having a driver in the home, hearing difficulties and incontinence were associated with low social contact; older age and African American race were related to home confinement. African American women living alone are especially vulnerable to home confinement. Physical disability is not necessarily socially disabling, as many of the most severely disabled in our study had at least daily social interaction. Improvements in social interaction appear possible through more effective management of certain health conditions and attention to potential sociocultural barriers.


[The structure of the centromeric region and chromosome behavior in the anaphase after incubating murine cells with the fluorochrome Hoechst 33258].
Mouse chromosome behaviour has been studied in the anaphase, after the cells (culture L) were incubated with fluorochrome Hoechst 33258, provoking the stretching of the chromosome centromeric regions. It was shown that in spite of marked structural changes in the centromere, the cells entered the anaphase and chromosomes preserved their ability to separate towards the spindle poles. At the same time, in the presence of Hoechst 33258, a reliable increase in the anaphase duration could be observed from 4.0 +/- 0.2 min in control to 6.0 +/- 0.5 min after a prolonged incubation of cells with the fluorochrome. Also, the increase in the frequency of chromosomal bridges in anaphase and the appearance of bridges in interphase cells were observed.


Prognostic implication of activated partial thromboplastin time after reteplase or half-dose reteplase plus abciximab: results from the GUSTO-V trial.
To evaluate the relationship between activated partial thromboplastin time (aPTT) and clinical outcomes in the Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO-V) trial comparing standard-dose reteplase to half-dose reteplase and abciximab. We analysed data on 11,420 patients receiving unfractionated heparin. Peak aPTT levels recorded during the hospitalization were correlated with clinical outcomes. Multivariable logistic regression models examined the relationship between peak aPTT levels and (i) moderate-to-severe bleeding, (ii) intracerebral haemorrhage, (iii) reinfarction, and (iv) 30-day mortality. Non-linear relationships were explored in the models using cubic spline functions. Higher rates of significant complications were seen in both groups when aPTT levels were <50 s or when levels were >70 s. In the combination therapy group, the relationship between aPTT levels and bleeding appeared accentuated. Reinfarction rates increased gradually as aPTT levels were >70 s in both groups, but the relationships were not statistically significant. Peak aPTT levels <50 s were associated with increased 30-day mortality even after multivariable adjustment. Peak aPTT levels <50 s and >70 s are associated with worse clinical outcomes in the modern era of fibrinolytic therapy; these relationships are different in patients receiving standard reteplase vs. combination therapy.


[Infectious spondylodiscitis: the surgical approach].
Introduction of antibiotics in the last century and recent progress in surgical techniques have significantly reduced mortality and morbidity rates of spondylodiscitis. However, when unrecognized, it can have catastrophic consequences for the patient. Antibiotic drugs remain the cornerstone of treatment, and surgery is to be discussed in general as a last resource in case of acute neurologic deficit or severe sagittal or frontal spinal imbalance. Metallic implants are not absolutely contraindicated in surgical treatment of spondylodiscitis and relapse of infection has been reported to be rare. In case of severe mechanical instability and neurologic deficit, they even are indispensable and complications can be avoided by accurately identifying the germ and by treating infection with the adequate antibiotic beforehand.


To integrate or not to integrate: Temporal dynamics of hierarchical Bayesian causal inference.
To form a percept of the environment, the brain needs to solve the binding problem-inferring whether signals come from a common cause and are integrated or come from independent causes and are segregated. Behaviourally, humans solve this problem near-optimally as predicted by Bayesian causal inference; but the neural mechanisms remain unclear. Combining Bayesian modelling, electroencephalography (EEG), and multivariate decoding in an audiovisual spatial localisation task, we show that the brain accomplishes Bayesian causal inference by dynamically encoding multiple spatial estimates. Initially, auditory and visual signal locations are estimated independently; next, an estimate is formed that combines information from vision and audition. Yet, it is only from 200 ms onwards that the brain integrates audiovisual signals weighted by their bottom-up sensory reliabilities and top-down task relevance into spatial priority maps that guide behavioural responses. As predicted by Bayesian causal inference, these spatial priority maps take into account the brain's uncertainty about the world's causal structure and flexibly arbitrate between sensory integration and segregation. The dynamic evolution of perceptual estimates thus reflects the hierarchical nature of Bayesian causal inference, a statistical computation, which is crucial for effective interactions with the environment.


Year-round screening of noncommercial and commercial oysters for the presence of human pathogenic viruses.
Consumption of virus-contaminated shellfish has caused numerous outbreaks of gastroenteritis and hepatitis worldwide. In The Netherlands, oysters are cultured and imported both for consumption and export; therefore, the presence of noroviruses, rotaviruses, astroviruses, hepatitis A viruses, and enteroviruses was determined in 64 commercial and noncommercial oyster samples. Oysters were collected monthly for 13 months from four different harvesting areas in the Oosterschelde Delta. Oyster samples were classified by determining Escherichia coli levels according to the standards set by the Councils Directive (91/ 492/EEC). Two of 36 commercial and 2 of 28 noncommercial oyster samples were B-classified and therefore not ready for consumption. All other oyster samples were A-classified. For the detection of viral RNA, 150 mg of hepatopancreatic tissue was subjected to the Qiagen RNeasy Mini Kit, followed by reverse transcriptase (RT)-PCR and Southern blot hybridization. Enterovirus RNA was detected in 14 of 64 oyster samples, of which 4 were from noncommercial oyster harvesting areas and 10 were from commercial harvesting areas. None of the other human pathogenic viruses were detected. The levels of somatic coliphages and F-specific phages were also determined in all 64 oyster samples, with some samples containing high phage levels (>50 PFU/g of hepatopancreatic tissue), but with most samples containing low phage levels (<50 PFU/g of hepatopancreatic tissue). However, independent of these high or low phage levels, enterovirus RNA could be detected. Thus, commercial oysters can be contaminated with pathogenic viruses, and monitoring only fecal indicators might not sufficiently protect human health.


Implementing the CanMEDSTM physician roles in rural specialist education: the Multi-Speciality Community Training Network.
Changing medical education to realign it with societal needs has become a renewed priority in many countries. Advanced training in rural settings to prepare physicians to better serve rural areas has received particular attention around the world. Such initiatives are usually targeted at primary care practitioners. Few initiatives have been designed to enhance specialist training in a rural setting, let alone adapt specialist competency frameworks such as the CanMEDSTM roles of the Royal College of Physicians and Surgeons of Canada to non-urban medical education. We describe an innovation in medical training for rural competence for specialist physicians using the CanMEDS framework near London, Ontario, Canada. Since 1997, the University of Western Ontario has established its Multi-Specialty Community Training Network (MSCTN) to provide rural and regional training opportunities for specialty residents in anaesthesia, general surgery, internal medicine, paediatrics, obstetrics and psychiatry. It became the first program in Canada to fully adapt the new CanMEDS roles into learning objectives and evaluations. Competency-based frameworks like CanMEDS are important because they provide a comprehensive tool to organize outcome-based curricula. The CanMEDS roles framework has been very useful in developing educational goals for rural/regional specialty resident rotations as well as forming a constructive basis for resident, preceptor, and program evaluations. Our experiences with this program may provide lessons for others planning training for specialists in rural settings, and those adopting the CanMEDS competency framework.


Development of a theoretical model for the inhibition of nsP3 protease of Chikungunya virus using pyranooxazoles.
Chikungunya virus (CHIKV) causes Chikungunya fever (CHIKF) and till date no effective medicine for its cure is available in market. Different research groups find various possible interactions between small molecules and non-structural proteins, viz. nsP3, one of the most important viral elements in CHIKV. In this work, authors have studied the interactions of nsP3 protease of CHIKV with pyranooxazoles. Initially, a one-pot three-component reaction was designed using oxazolidine-2,4-dione, benzaldehyde and cyanoethylacetate to get a proposed biological active molecule, i.e. based on pyranooxazoles. The mechanism for the synthesis of the product based on pyranooxazole was studied through density functional theory (DFT) using Gaussian. Then, a library of the obtained pyranooxazole was created through computational tools by varying the substituents. Further, virtual screening of the designed library of pyranooxazoles (200 compounds) against nsP3 protease of CHIKV was performed. Herein, CMPD 104 showed strongest binding affinity toward the targeted nsP3 protease of CHIKV, based on the least binding energy obtained from docking. Based on docking results, the pharmacological, toxicity, biological score and Lipinski's filters were studied. Further, DFT studies of top five compounds were done using Gaussian. Molecular dynamics (MD) simulation of nsP3 protease of CHIKV with and without 104 was performed using AMBER18 utilizing ff14SB force field in three steps (minimization, equilibration and production). This work is emphasized to designing of one-pot three-component synthesis and to develop a theoretical model to inhibit the nsP3 protease of CHIKV. AbbreviationsCHIKFChikungunya feverCHIKVChikungunya virusDFTdensity functional theoryDSDiscovery StudioMDmolecular dynamicsMM-GBSAmolecular mechanics-generalized born surface areaMMVMolegro molecular viewerCommunicated by Ramaswamy H. Sarma.


Acetylation of FoxO1 activates Bim expression to induce apoptosis in response to histone deacetylase inhibitor depsipeptide treatment.
Histone deacetylase (HDAC) inhibitors have been shown to induce cell cycle arrest and apoptosis in cancer cells. However, the mechanisms of HDAC inhibitor induced apoptosis are incompletely understood. In this study, depsipeptide, a novel HDAC inhibitor, was shown to be able to induce significant apoptotic cell death in human lung cancer cells. Further study showed that Bim, a BH3-only proapoptotic protein, was significantly upregulated by depsipeptide in cancer cells, and Bim's function in depsipeptide-induced apoptosis was confirmed by knockdown of Bim with RNAi. In addition, we found that depsipeptide-induced expression of Bim was directly dependent on acetylation of forkhead box class O1 (FoxO1) that is catalyzed by cyclic adenosine monophosphate-responsive element-binding protein-binding protein, and indirectly induced by a decreased four-and-a-half LIM-domain protein 2. Moreover, our results demonstrated that FoxO1 acetylation is required for the depsipeptide-induced activation of Bim and apoptosis, using transfection with a plasmid containing FoxO1 mutated at lysine sites and a luciferase reporter assay. These data show for the first time that an HDAC inhibitor induces apoptosis through the FoxO1 acetylation-Bim pathway.


Selective therapeutic control of C5a and the terminal complement complex by anti-C5 single-chain Fv in an experimental model of antigen-induced arthritis in rats.
To determine the role of the terminal complement complex (TCC) in the development of experimental antigen-induced arthritis (AIA) and the therapeutic effects of human anti-C5 single-chain Fv (scFv). Two different anti-C5 scFv, one that inhibits both release of C5a and assembly of the TCC (TS-A 12/22) and another that selectively blocks formation of the TCC (TS-A 8), were injected at the onset of AIA. The effects of these scFv on disease severity were evaluated for up to 21 days and compared with the effects of injection of an unrelated scFv. AIA was also established in C6-deficient and C6-sufficient PVG rats to obtain further information on the role of the TCC in this model. TS-A 12/22 and TS-A 8 proved to be equally effective in reducing joint swelling, cell counts and tumor necrosis factor alpha levels in synovial lavage fluids, and the degree of histomorphologic changes compared with the effects of the unrelated scFv. TS-A 12/22 and TS-A 8 prevented the deposition of C9 but not that of C3, confirming the ability of the 2 scFv to neutralize C5. Administration of the 2 anti-C5 scFv after AIA onset also reduced disease severity. In C6-deficient rats with AIA, disease activity was reduced markedly compared with that in C6-sufficient rats. These 2 human anti-C5 scFv could represent potential therapeutic reagents to be used in patients with rheumatoid arthritis. In addition, the finding that TS-A 8 was as effective as TS-A 12/22 in reducing disease severity suggests that the TCC is mainly responsible for the joint inflammation and damage observed in AIA.


The process of curriculum development and implementation for an adolescent health project in middle schools.
Childhood obesity has reached epidemic levels in developed countries and is showing no signs of abating. The causes of obesity in adolescence are extremely complex, and therefore approaches to prevention and treatments must be multifaceted. Early adolescence is a developmental period when youth are becoming more independent, are influenced by peers, and are making more decisions related to their own health and lifestyles. The purpose of this article is to describe the development, implementation, and evaluation of a multilevel school-based intervention program for middle school students in the Teen Eating and Activity Mentoring in Schools (TEAMS) project. Critical components of success included an interdisciplinary approach, applying available curricula to meet contextual characteristics, and involvement of all stakeholders in planning, evaluating, and refining the program.


Multi-scale X-ray computed tomography to detect and localize metal-based nanomaterials in lung tissues of in vivo exposed mice.
In this methodological study, we demonstrated the relevance of 3D imaging performed at various scales for the ex vivo detection and location of cerium oxide nanomaterials (CeO2-NMs) in mouse lung. X-ray micro-computed tomography (micro-CT) with a voxel size from 14 µm to 1 µm (micro-CT) was combined with X-ray nano-computed tomography with a voxel size of 63 nm (nano-CT). An optimized protocol was proposed to facilitate the sample preparation, to minimize the experimental artifacts and to optimize the contrast of soft tissues exposed to metal-based nanomaterials (NMs). 3D imaging of the NMs biodistribution in lung tissues was consolidated by combining a vast variety of techniques in a correlative approach: histological observations, 2D chemical mapping and speciation analysis were performed for an unambiguous detection of NMs. This original methodological approach was developed following a worst-case scenario of exposure, i.e. high dose of exposure with administration via intra-tracheal instillation. Results highlighted both (i) the non-uniform distribution of CeO2-NMs within the entire lung lobe (using large field-of-view micro-CT) and (ii) the detection of CeO2-NMs down to the individual cell scale, e.g. macrophage scale (using nano-CT with a voxel size of 63 nm).


Motion-Binding Property Contributes to Accurate Temporal-Order Perception in Audiovisual Synchrony.
Temporal perception in multisensory processing is important for an accurate and efficient understanding of the physical world. In general, it is executed in a dynamic environment in our daily lives. In particular, the motion-binding property is important for correctly identifying moving objects in the external environment. However, how this property affects multisensory temporal perception remains unclear. We investigate whether the motion-binding property influences audiovisual temporal integration. The study subjects performed four types of temporal-order judgment (TOJ) task experiments using three types of perception. In Experiment 1, the subjects conducted audiovisual TOJ tasks in the motion-binding condition, between two flashes, and in the simultaneous condition, in which the two flashes are perceived as simultaneous stimuli without motion. In Experiment 2, subjects conducted audiovisual TOJ tasks in the motion-binding condition and the short and long successive interval condition, in which the two stimuli are perceived as successive with no motion. The results revealed that the point of subjective simultaneity (PSS) and the just-noticeable difference (JND) in the motion-binding condition differed significantly from those in the simultaneous and short and long successive interval conditions. Specifically, the PSS in the motion-binding condition was shifted toward a sound-lead stimulus in which the PSS became closer to zero (i.e., physical simultaneity) and the JND became narrower compared to other conditions. This suggests that the motion-binding property contributes to accurate temporal integration in multisensory processing by precisely encoding the temporal order of the physical stimuli.


Testing the stand-alone microbeam at Columbia University.
The stand-alone microbeam at Columbia University presents a novel approach to biological microbeam irradiation studies. Foregoing a conventional accelerator as a source of energetic ions, a small, high-specific-activity, alpha emitter is used. Alpha particles emitted from this source are focused using a compound magnetic lens consisting of 24 permanent magnets arranged in two quadrupole triplets. Using a 'home made' 6.5 mCi polonium source, a 1 alpha particle s(-1), 10 microm diameter microbeam can, in principle, be realised. As the alpha source energy is constant, once the microbeam has been set up, no further adjustments are necessary apart from a periodic replacement of the source. The use of permanent magnets eliminates the need for bulky power supplies and cooling systems required by other types of ion lenses and greatly simplifies operation. It also makes the microbeam simple and cheap enough to be realised in any large lab. The Microbeam design as well as first tests of its performance, using an accelerator-based beam are presented here.


Analysis and Adjustment of Vertebral Subluxation as a Separate and Distinct Identity for the Chiropractic Profession: A Commentary.
The purpose of this paper is to discuss various statements related to chiropractic identity from1 D. D. Palmer2; selected chiropractic organizations, associations, and colleges; and3 attitudes and perceptions of chiropractic from chiropractic students, practitioners, and patients. For comparison purposes, identity statements and perceptions from the various chiropractic associations and colleges, as well as from students and patients, were explored. Identity statements for chiropractic were searched in various sources such as Palmer's 1910 textbook, recent literature on viewpoints from chiropractic students and practitioners, and websites for chiropractic colleges and organizations. Palmer taught that the chiropractor's focus was on vertebral subluxation. Today, a number of chiropractic colleges and organizations continue to include the vertebral subluxation model in their instruction, with a majority of students and practitioners subscribing to the model. Conversely, a number of other colleges and organizations portray chiropractic as being essentially about the treatment of back and neck pain, which is what patients associate with chiropractic. However, settling on any particular identity for the chiropractic profession will likely be met with resistance by some, given the plethora of opinions among chiropractic professionals as to what the identity of the chiropractic profession should be. Common ground between the different factions within the chiropractic profession might be found in a unifying expression such as "functional neurology." When a profession's identity is not clear with respect to its area of interest and mission, then the public may be less inclined to seek its services. Identifying the chiropractic profession with a focus on vertebral subluxation would give the profession uniqueness not duplicated by other health care professions and, therefore, might legitimatize the existence of chiropractic as a health care profession. An identity having a focus on vertebral subluxation would also be consistent with the original intent of the founding of the chiropractic profession.


Transcriptome Profiling of Long Non-coding RNAs During the Atlantic Salmon Smoltification Process.
For salmon aquaculture, one of the most critical phase is the parr-smolt transformation. Studies around this process have mainly focused on physiological changes and the Na+/K+-ATPase activity during the osmoregulatory activity. However, understanding how the salmon genome regulates the parr-smolt transformation, specifically the molecular mechanisms involved, remains uncovered. This study aimed to explore the transcriptional modulation of long non-coding RNAs (lncRNAs), as key molecular regulators, during the freshwater (FW) to seawater (SW) transfer in Atlantic salmon. Transcriptome sequencing was performed from gill samples of Atlantic salmon adapted from FW to SW through gradual salinity changes from 0 to 30 PSU. The results showed that most transcripts differently modulated were downregulated in all salinity conditions. Relevant biological processes were associated with growth, collagen formation, immune response, metabolism, and heme transport. Notably, 2864 putative lncRNAs were identified in Atlantic salmon gills differently expressed during fish smoltification. The highest number of lncRNAs differently modulated was observed at 30 PSU. Correlation expression analysis suggests putative regulatory roles of lncRNAs with smoltification-related genes. Herein, co-localization of Na+/K+-ATPase, growth hormone receptor, and thyroid hormone receptor genes with lncRNAs differentially expressed suggest putative regulatory mechanisms in the Atlantic salmon genome. The lncRNAs can be used as novel biomarkers for the fish smoltification process. Here, the lncRNA_145326 and lncRNA_18762 are putatively related to the parr-smolt transfer in Atlantic salmon. This study is the first description of lncRNAs with putative regulatory roles in Atlantic salmon during the SW adaptation.


Evaluation of environmental contaminations and occupational exposures involved in preparation of chemotherapeutic drugs.
Many healthcare workers are concerned about the risk of occupational exposures to hazardous drugs. The Japanese Society of Hospital Pharmacists (JSHP) revised the "Guidelines for the Handling of Antineoplastic Drugs in Hospitals", however, the precautions and awareness of handling drugs varied in institutions. We assessed the levels of environmental contaminations in our hospital and urinary excretion of cyclophosphamide (CP) and ifosfamide (IF) in pharmacists and nurses. In environmental studies, we obtained samples by wiping the surfaces around two biological safety cabinets (BSCs) on eight days for four months. One BSC was equipped in hospital pharmacy and the other was equipped in an oncology ward, and used for preparing chemotherapeutic drugs for outpatients and for inpatients, respectively. We obtained the urine samples from 6 pharmacists and 2 nurses. We used solid phase extraction (SPE) as a convenient extraction procedure and liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) for the analysis of the samples. CP was detected on the working surfaces inside both BSCs, and detected at low levels on the back surfaces of the BSCs and at the working tables around the BSCs. IF over the LLOQ was not detected in both BSCs. CP and IF were not detected in all urine samples of pharmacists and nurses. Detection frequencies and amounts of these drugs were low levels, compared with previous reports in Japan, and our results showed that improving awareness about handling hazardous drugs could reduce the risk of the occupational exposures.


Why twenty amino acid residue types suffice(d) to support all living systems.
It is well known that proteins are built up from an alphabet of 20 different amino acid types. These suffice to enable the protein to fold into its operative form relevant to its required functional roles. For carrying out these allotted functions, there may in some cases be a need for post-translational modifications and it has been established that an additional three types of amino acid have at some point been recruited into this process. But it still remains the case that the 20 residue types referred to are the major building blocks in all terrestrial proteins, and probably "universally". Given this fact, it is surprising that no satisfactory answer has been given to the two questions: "why 20?" and "why just these 20?". Furthermore, a suggestion is made as to how these 20 map to the codon repertoire which in principle has the capacity to cater for 64 different residue types. Attempts are made in this paper to answer these questions by employing a combination of quantum chemical and chemoinformatic tools which are applied to the standard 20 amino acid types as well as 3 "non-standard" types found in nature, a set of fictitious but feasible analog structures designed to test the need for greater coverage of function space and the collection of candidate alternative structures found either on meteorites or in experiments designed to reconstruct pre-life scenarios.


Valsartan slows the progression of diabetic nephropathy in db/db mice via a reduction in podocyte injury, and renal oxidative stress and inflammation.
Higher doses of AngII (angiotensin II) blockers are intended to optimize albuminuria reduction rather than for blood pressure control in chronic kidney disease. However, the long-term renoprotection of high-dose AngII blockers has yet to be defined. The present study sought to determine whether doses of ARB (AngII receptor blocker) that maximally reduce proteinuria could slow the progression of glomerulosclerosis in the uninephrectomized db/db mouse, a model of Type 2 diabetes. Untreated uninephrectomized db/db mice had normal blood pressure, but developed progressive albuminuria and mesangial matrix expansion between 18 and 22 weeks of age, which was associated with increased renal expression of TGFβ1 (transforming growth factor β1), PAI-1 (plasminogen-activator inhibitor-1), type IV collagen and FN (fibronectin). Treatment with valsartan in the drinking water of db/db mice from 18 to 22 weeks of age, at a dose that was determined previously to maximally reduce proteinuria, prevented the increases in albuminuria and the markers of renal fibrosis seen in untreated db/db mice. In addition, WT-1 (Wilms tumour protein-1)-immunopositive podocyte numbers were found to be lower in the untreated glomeruli of mice with diabetes. The expression of podocin and nephrin were continually decreased in mice with diabetes between 18 and 22 weeks of age. These changes are indicative of podocyte injury and the administration of valsartan ameliorated them substantially. Renal expression of TNFα (tumour necrosis factor α), MCP-1 (monocyte chemoattractant protein-1), Nox2 (NADPH oxidase 2), p22phox and p47phox and urine TBARS (thiobarbituric acid-reacting substance) levels, the markers of renal inflammation and oxidative stress, were increased during disease progression in mice with diabetes. Valsartan treatment was shown to reduce these markers. Thus high doses of valsartan not only reduce albuminuria maximally, but also halt the progression of the glomerulosclerosis resulting from Type 2 diabetes via a reduction in podocyte injury and renal oxidative stress and inflammation.


Intratracheal bleomycin causes airway remodeling and airflow obstruction in mice.
In addition to parenchymal fibrosis, fibrotic remodeling of the distal airways has been reported in interstitial lung diseases. Mechanisms of airway wall remodeling, which occurs in a variety of chronic lung diseases, are not well defined and current animal models are limited. The authors quantified airway remodeling in lung sections from subjects with idiopathic pulmonary fibrosis (IPF) and controls. To investigate intratracheal bleomycin as a potential animal model for fibrotic airway remodeling, the authors evaluated lungs from C57BL/6 mice after bleomycin treatment by histologic scoring for fibrosis and peribronchial inflammation, morphometric evaluation of subepithelial connective tissue volume density, TUNEL (terminal deoxynucleotidyl transferase dUTP-mediated nick-end labeling) assay, and immunohistochemistry for transforming growth factor β1 (TGFβ1), TGFβ2, and the fibroblast marker S100A4. Lung mechanics were determined at 3 weeks post bleomycin. IPF lungs had small airway remodeling with increased bronchial wall thickness compared to controls. Similarly, bleomycin-treated mice developed dose-dependent airway wall inflammation and fibrosis and greater airflow resistance after high-dose bleomycin. Increased TUNEL(+) bronchial epithelial cells and peribronchial inflammation were noted by 1 week, and expression of TGFβ1 and TGFβ2 and accumulation of S100A4(+) fibroblasts correlated with airway remodeling in a bleomycin dose-dependent fashion. IPF is characterized by small airway remodeling in addition to parenchymal fibrosis, a pattern also seen with intratracheal bleomycin. Bronchial remodeling from intratracheal bleomycin follows a cascade of events including epithelial cell injury, airway inflammation, profibrotic cytokine expression, fibroblast accumulation, and peribronchial fibrosis. Thus, this model can be utilized to investigate mechanisms of airway remodeling.


Percolation threshold of graphene nanosheets as conductive additives in Li4Ti5O12 anodes of Li-ion batteries.
Graphene nanosheets (GNSs) have been considered as potential conductive additives for electrodes in Li-ion batteries to replace the existing carbon black (CB). Graphene has exceptionally high aspect ratio and excellent electrical conductivity, enabling the formation of extensive conductive networks at a much lower content than CB. This paper reports the beneficial effects of GNSs with a low percolation threshold on electrochemical performance of Li(4)Ti(5)O(12) (LTO) anodes. The experimental results show that the GNSs with a diameter of 46 μm and a thickness of 4.5 nm have a percolation threshold of 1.8 wt%. The prediction based on the interparticle distance concept gives a percolation threshold of 0.54 wt% for GNSs, which is almost an order of magnitude lower than that for CB particles. The substantially low percolation along with a high electrical conductivity of GNSs explains why the LTO anodes containing only 5 wt% GNSs deliver a much better rate capability than those with 15 wt% CB. However, a higher GNS content of 10 wt% results in re-stacking GNSs, deteriorating the diffusion of Li ions through the thickness of GNSs. The parametric study indicates that the percolation threshold of GNSs is inversely proportional to the aspect ratio of GNSs.


Microbially-driven strategies for bioremediation of bauxite residue.
Globally, 3 Gt of bauxite residue is currently in storage, with an additional 120 Mt generated every year. Bauxite residue is an alkaline, saline, sodic, massive, and fine grained material with little organic carbon or plant nutrients. To date, remediation of bauxite residue has focused on the use of chemical and physical amendments to address high pH, high salinity, and poor drainage and aeration. No studies to date have evaluated the potential for microbial communities to contribute to remediation as part of a combined approach integrating chemical, physical, and biological amendments. This review considers natural alkaline, saline environments that present similar challenges for microbial survival and evaluates candidate microorganisms that are both adapted for survival in these environments and have the capacity to carry out beneficial metabolisms in bauxite residue. Fermentation, sulfur oxidation, and extracellular polymeric substance production emerge as promising pathways for bioremediation whether employed individually or in combination. A combination of bioaugmentation (addition of inocula from other alkaline, saline environments) and biostimulation (addition of nutrients to promote microbial growth and activity) of the native community in bauxite residue is recommended as the approach most likely to be successful in promoting bioremediation of bauxite residue.


Relationship of desipramine plasma levels to therapeutic response: a critical reappraisal of the data.
The literature on the relationship between desipramine plasma levels and therapeutic response is reviewed. Although no clear pattern has emerged from the available studies, the data suggest that plasma levels of 125 ng/ml should be attained before moderately to severely ill patients are considered nonresponsive to desipramine. An upper therapeutic plasma level of 170 ng/ml may exist for patients with less severe illness. Future studies should control for severity of illness in assessing dose-response relationships.


A General Approach for Generating Fluorescent Probes to Visualize Piconewton Forces at the Cell Surface.
Mechanical forces between cells and their extracellular matrix (ECM) are mediated by dozens of different receptors. These biophysical interactions play fundamental roles in processes ranging from cellular development to tumor progression. However, mapping the spatial and temporal dynamics of tension among various receptor-ligand pairs remains a significant challenge. To address this issue, we have developed a synthetic strategy to generate modular tension probes combining the native chemical ligation (NCL) reaction with solid phase peptide synthesis (SPPS). In principle, this approach accommodates virtually any peptide or expressed protein amenable to NCL. We generated a small library of tension probes displaying different ligands, flexible linkers, and fluorescent reporters, enabling the mapping of integrin and cadherin tension, and demonstrating the first example of long-term (∼3 days) molecular tension imaging. This approach provides a toolset to better understand mechanotransduction events fundamental to cell biology.


Suspected exercise-induced seizures in a young dog.
A 12-month-old female neutered crossbreed was referred for investigation of seizure-like episodes occurring only at intense exercise. Thorough medical, neurological and cardiac investigations were performed and excluded the most commonly known causes of seizure-like activity. The dog was fitted with an ambulatory electrocardiography device and underwent another exercise-induced seizure. The electrocardiogram during the episode revealed a sinus tachycardia at approximately 300 beats/minute. A video recording of the episode revealed generalised tonic clonic limb activity with jaw chomping and frothing at the mouth typical of seizure activity. Antiepileptic medications were not prescribed and the owner was advised not to exercise the dog intensely. The dog responded well and did not seizure after 12 months of mild-moderate off-lead exercise. As all the seizures in this case were triggered by intense physical activity, it is suggested that this may be a new form of reflex seizure activity.


Lipoxygenase-catalyzed oxidation of catecholamines.
Dopa and structurally related catecholamines in presence of hydrogen peroxide are oxidized in vitro by soybean lipoxygenase producing the corresponding melanin pigments. The kinetic parameters of the catecholasic reaction, measured as aminochrome formation, have been calculated. The rate of peroxidation depends on catecholamine and hydrogen peroxide concentration. The optimum pH for the peroxidative activity of the enzyme is around 8.5. The enzyme, at higher pH values (pH 9-9.5), is also able to perform an oxidative reaction of the substrates. Implications of the possible biochemical relevance of the reactions are discussed.


[Treatment of asthma in children; revised guidelines by pediatric pneumologists. Section of Pediatric Lung Diseases of the Dutch Association of Pediatric Medicine].
The diagnosis of 'asthma' is still based on anamnesis, physical examination and lung function tests. Supplementary examinations (laboratory or roentgen) are carried out only if indicated. Most young children with recurrent periods of wheezing and coughing prove not to develop asthma in the long run. Oral medication no longer has a place in the treatment of asthma, since inhalation medication is now possible at all ages, among other things because of the current availability of new nozzle attachments. Every child with asthma should have a short-acting beta 2-agonist at hand. Furthermore, inhaled corticosteroids are the maintenance drug of choice in children with moderate or severe asthma. Systemic effects may occur with every inhaled corticosteroid, even with dosages usual for children, but these are rarely relevant. Cromones nowadays play a very limited part. Long-acting beta 2-agonists are indicated in children whose asthma cannot be controlled with standard doses of inhaled corticosteroids. In the prevention of allergic asthma of childhood, prescription of the allergen-tight mattress cover plays a main part. The physician has an important task in discouraging (passive) smoking. Young children who, in spite of treatment with inhaled corticosteroids, have recurrent episodes of wheezing and coughing. and children requiring high doses of inhaled corticosteroids (over 400 micrograms beclomethasone/budesonide or 250 micrograms fluticasone) should be referred to a paediatrician. The asthma nurse has an important task in instructing and advising the asthmatic child and its parents. Self-management programmes may be of great psychological value for the patient and his environment.


In vivo protective effect of lipopolysaccharide against Pseudomonas aeruginosa exotoxin A in mice.
Lipopolysaccharide (LPS) treatment of mice 1 to 5 days prior to administration of Pseudomonas aeruginosa exotoxin A (PA) induced full or partial protection against PA intoxication. The optimal LPS dose that induced resistance was 50 to 100 micrograms per mouse. Simultaneous administration of LPS and PA to mice, however, increased their sensitivity to PA two- to fourfold. Mice pretreated with LPS demonstrated a markedly enhanced clearance rate of 125I-labeled PA from peripheral blood, livers, and kidneys. In mice exposed to LPS and PA simultaneously, the rate of elimination of labeled PA was lower than that in control mice. While protein synthesis was inhibited significantly in livers and other organs of PA-exposed mice, in LPS-pretreated mice, PA-induced inhibition of protein synthesis was either diminished or totally prevented and elongation factor 2 (EF2) levels were normal. In mice treated only with LPS, enhanced protein synthesis and increased levels of EF2 were observed, suggesting that LPS protection against PA intoxication was perhaps a consequence of excessive amounts of EF2 induced by LPS.


Optimized homomorphic unsharp masking for MR grayscale inhomogeneity correction.
Grayscale inhomogeneities in magnetic resonance (MR) images confound quantitative analysis of these images. Homomorphic unsharp masking and its variations have been commonly used as a post-processing method to remove inhomogeneities in MR images. However, little data is available in the literature assessing the relative effectiveness of these algorithms to remove inhomogeneities, or describing how these algorithms can affect image data. In this study, we address these questions quantitatively using simulated images with artificially constructed and empirically measured bias fields. Our results show that mean-based filtering is consistently more effective than median-based algorithms for removing inhomogeneities in MR images, and that artifacts are frequently introduced into images at the most commonly used window sizes. Our results demonstrate dramatic improvement in the effectiveness of the algorithms with significantly larger windows than are commonly used.


Clinical Laboratory Tests Used To Aid in Diagnosis of Human Prion Disease.
Prion diseases are a group of rapidly progressive and always fatal neurodegenerative disorders caused by misfolded prion protein in the brain. Although autopsy remains the gold-standard diagnostic tool, antemortem laboratory testing can be performed to aid in the diagnosis of prion disease. This review is meant to help laboratory directors and physicians in their interpretation of test results. Laboratory assays to detect both nonspecific biomarkers of prion disease and prion-specific biomarkers can be used. The levels of nonspecific biomarkers in cerebrospinal fluid (CSF) are elevated when rapid neurodegeneration is occurring in the patient, and these markers include 14-3-3, tau, neuron-specific enolase, S100B, and alpha-synuclein. These markers have various sensitivities and specificities but are overall limited, as the levels of any of these analytes can be elevated in nonprion disease that is causing rapid damage of brain tissue. Prion-specific assays used in clinical laboratory testing are currently limited to two options. The first option is second-generation real-time quaking-induced conversion (RT-QuIC) performed on CSF, and the second option is Western blotting of a brain biopsy specimen used to detect protease-resistant prion protein. Although both tests have exquisite specificity, RT-QuIC has a sensitivity of 92 to 97.2% in symptomatic individuals, compared to the brain biopsy Western blot sensitivity of 20 to 60%. RT-QuIC was added to the Centers for Disease Control and Prevention's diagnostic criteria for prion disease in 2018. Other caveats of laboratory testing need to be considered, as sporadic, genetic, and acquired forms of prion disease have different clinical and laboratory presentations, and these caveats are discussed. Laboratory testing plays an important role in the diagnosis of prion disease, which is often challenging to diagnose.


Meta-analysis of Postoperative Mortality and Morbidity After Total Abdominal Colectomy Versus Loop Ileostomy With Colonic Lavage for Fulminant Clostridium Difficile Colitis.
Emergency surgery is often required for fulminant Clostridium difficile colitis. Total abdominal colectomy has been the treatment of choice despite high morbidity and mortality. The aim of this meta-analysis was to evaluate postoperative mortality and morbidity after total abdominal colectomy and loop ileostomy with colonic lavage in patients with fulminant C difficile colitis. Studies comparing total abdominal colectomy to loop ileostomy for fulminant C difficile colitis were identified by a systematic search of PubMed, Cochrane Library, MEDLINE, and CINAHL. Relevant records were detected and screened using a cascade system (title, abstract, and/or full text article). Total abdominal colectomy (rectal-sparing resection of the entire colon with end ileostomy) was compared to loop ileostomy (exteriorization of an ileal loop not far from the ileocecal junction for colonic lavage). This meta-analysis was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Guidelines. Primary outcome was postoperative mortality, defined as death occurring within 30 days after the intervention. Secondary end points were the rates of ostomy reversal, deep venous thrombosis/embolism, surgical site infection, urinary tract infection, respiratory complications, reoperations, and adverse events. Mantel-Haenszel method with random-effects model was used for meta-analysis. Five observational studies (3 cohort and 2 database analysis studies) totaling 3683 patients were included. Postoperative mortality rate was 31.3% after total abdominal colectomy and 26.2% after loop ileostomy (OR = 1.36 (95% CI, 0.83-2.24); p = 0.22; number needed to treat/harm = 20; I = 55%). Ostomy reversal rate was both statistically and clinically significantly higher after loop ileostomy as compared with total abdominal colectomy (80% vs 25%; OR = 0.08 (95% CI, 0.02-0.30); p = 0.002; number needed to treat/harm = 2) with low heterogeneity (I = 0%). A limitation is the observational nature of the included studies introducing an overall high risk of selection bias. This meta-analysis suggests that loop ileostomy with colonic lavage for fulminant C difficile colitis may be associated with similar survival and decreased surgical site infection rates as compared with total abdominal colectomy. Although loop ileostomy with colonic lavage was associated with higher ostomy reversal rates, this finding was based on the data from only 2 studies.


Development of a cytokine analog with enhanced stability using computational ultrahigh throughput screening.
Granulocyte-colony stimulating factor (G-CSF) is used worldwide to prevent neutropenia caused by high-dose chemotherapy. It has limited stability, strict formulation and storage requirements, and because of poor oral absorption must be administered by injection (typically daily). Thus, there is significant interest in developing analogs with improved pharmacological properties. We used our ultrahigh throughput computational screening method to improve the physicochemical characteristics of G-CSF. Improving these properties can make a molecule more robust, enhance its shelf life, or make it more amenable to alternate delivery systems and formulations. It can also affect clinically important features such as pharmacokinetics. Residues in the buried core were selected for optimization to minimize changes to the surface, thereby maintaining the active site and limiting the designed protein's potential for antigenicity. Using a structure that was homology modeled from bovine G-CSF, core designs of 25-34 residues were completed, corresponding to 10(21)-10(28) sequences screened. The optimal sequence from each design was selected for biophysical characterization and experimental testing; each had 10-14 mutations. The designed proteins showed enhanced thermal stabilities of up to 13 degrees C, displayed five-to 10-fold improvements in shelf life, and were biologically active in cell proliferation assays and in a neutropenic mouse model. Pharmacokinetic studies in monkeys showed that subcutaneous injection of the designed analogs results in greater systemic exposure, probably attributable to improved absorption from the subcutaneous compartment. These results show that our computational method can be used to develop improved pharmaceuticals and illustrate its utility as a powerful protein design tool.


[Phospholipid content of the rat lung and surfactant during the administration of hydrocortisone or insulin and in hypoxic hypoxia].
After acute hypoxia (3 hrs, P-200 mm Hg) content of total phospholipids and of their fractions was not altered in rat lung parenchyma, but the phospholipid composition, changed due to preadministration of hydrocortisone or insulin, was normalized. After the hypoxia concentration of phospholipids in surfactant was decreased mainly due to a loss of phosphatidyl choline; the phenomenon was noted and after preadministration of the hormones.


Effects of a growth hormone pulse on total and forearm substrate fluxes in humans.
Under physiological circumstances growth hormone (GH) is secreted in bursts after the onset of sleep and a few hours postprandially. Because most relevant studies have employed constant or repeated infusion of high doses of GH, the possible metabolic effects of such bursts are largely unknown. We have studied seven healthy, male subjects for 7 h after an intravenous bolus of 1) 140 micrograms GH and 2) saline. When injected, serum GH rose to a peak of 21 +/- 3 micrograms/l 10 min after injection. GH caused 1) a rapid, sustained 55% decrease in forearm glucose uptake (P less than 0.05) followed by increases toward control values, 2) a delayed 5 mg/100 ml decrease in plasma glucose (P less than 0.05), and 3) significant 60-250% increases (P less than 0.05) in all measured lipid intermediates (nonesterified fatty acids, 3-hydroxybutyrate, and glycerol) 120-160 min after administration followed by decreases to below control values (P less than 0.05). GH did not influence circulating levels of insulin, C-peptide, glucagon, or insulin-like growth factor I (IGF-I), or isotopically determined glucose turnover. Physiological bursts of GH secretion appear to have acute insulin antagonistic effects with maximal effect on lipolysis after 2 h. These effects are reversed after 4 h. Therefore, GH could play a key role in regulation of diurnal rhythms of substrate levels and fuel utilization in humans.


Sodium and ultrafiltration profiling in hemodialysis: A long-forgotten issue revisited.
Sodium and ultrafiltration profiling are method of dialysis in which dialysate sodium concentration and ultrafiltration rate are altered during the course of the dialysis session. Sodium and ultrafiltration profiling have been used, commonly simultaneously, to improve hemodynamic stability during hemodialysis. Sodium profiling is particularly effective in decreasing the incidence of intradialytic hypotension, while ultrafiltration profiling is suggested to decrease subclinical repeated end organ ischemia during dialysis. However, complications such as increased interdialytic weight gain and thirst due to sodium excess have prevented widespread use of sodium profiling. Evidence suggest that different sodium profiling techniques may lead to different clinical results, and preferring sodium balance neutral sodium profiling may mitigate adverse effects related to sodium overload. However, evidence is lacking on the long-term clinical outcomes of different sodium profiling methods. Optimal method of sodium profiling as well as the utility of sodium/ultrafiltration profiling in routine practice await further clinical investigation.


Impact of Prolonged Fraction Delivery Time Modelling Stereotactic Body Radiation Therapy with High Dose Hypofractionation on the Killing of Cultured ACHN Renal Cell Carcinoma Cell Line.
Stereotactic body radiotherapy delivers hypofractionated irradiation with high dose per fraction through complex treatment techniques. The increased complexity leads to longer dose delivery times for each fraction. The purpose of this study is to investigate the impact of prolonged fraction delivery time with high-dose hypofractionation on the killing of cultured ACHN cells. The radiobiological characteristics and repair half-time of human ACHN renal cell carcinoma cell line were studied with clonogenic assays. A total dose of 20 Gy was administered in 1, 2 or 3 fractions over 15, 30 or 45 min to investigate the biological effectiveness of radiation delivery time and hypofractionation. Cell cycle and apoptosis analysis was performed after 3-fraction irradiation over 30 and 45 min. The α/β and repair half-time were 5.2 Gy and 19 min, respectively. The surviving fractions increased with increase in the fraction delivery time and decreased more pronouncedly with increase in the fraction number over a treatment period of 30 to 45 min. With increase in the total radiation time to 30 and 45 min, it was found that with the same total dose, 2- and 3-fraction irradiation led to more cell killing than 1-fraction irradiation. 3-fraction radiation induced G2/M arrest, and the percentage of apoptotic cells decreased when the fraction delivery time increased from 30 min to 45 min. Our findings revealed that sublethal damage repair and redistribution of the cell cycle were predominant factors affecting cell response in the prolonged and hypofractionated irradiation regimes, respectively.


Abortion Surveillance - United States, 2015.
Since 1969, CDC has conducted abortion surveillance to document the number and characteristics of women obtaining legal induced abortions in the United States. 2015. Each year, CDC requests abortion data from the central health agencies of 52 reporting areas (the 50 states, the District of Columbia, and New York City). The reporting areas provide this information voluntarily. For 2015, data were received from 49 reporting areas. Abortion data provided by these 49 reporting areas for each year during 2006-2015 were used in trend analyses. Census and natality data were used to calculate abortion rates (number of abortions per 1,000 women aged 15-44 years) and ratios (number of abortions per 1,000 live births), respectively. A total of 638,169 abortions for 2015 were reported to CDC from 49 reporting areas. Among these 49 reporting areas, the abortion rate for 2015 was 11.8 abortions per 1,000 women aged 15-44 years, and the abortion ratio was 188 abortions per 1,000 live births. From 2014 to 2015, the total number of reported abortions decreased 2% (from 652,639), the abortion rate decreased 2% (from 12.1 abortions per 1,000 women aged 15-44 years), and the abortion ratio decreased 2% (from 192 abortions per 1,000 live births). From 2006 to 2015, the total number of reported abortions decreased 24% (from 842,855), the abortion rate decreased 26% (from 15.9 abortions per 1,000 women aged 15-44 years), and the abortion ratio decreased 19% (from 233 abortions per 1,000 live births). In 2015, all three measures reached their lowest level for the entire period of analysis (2006-2015). In 2015 and throughout the period of analysis, women in their 20s accounted for the majority of abortions and had the highest abortion rates; women aged ≥30 years accounted for a smaller percentage of abortions and had lower abortion rates. In 2015, women aged 20-24 and 25-29 years accounted for 31.1% and 27.6% of all reported abortions, respectively, and had abortion rates of 19.9 and 17.9 abortions per 1,000 women aged 20-24 and 25-29 years, respectively. In contrast, women aged 30-34, 35-39, and ≥40 years accounted for 17.7%, 10.0%, and 3.5% of all reported abortions, respectively, and had abortion rates of 11.6, 7.0, and 2.5 abortions per 1,000 women aged 30-34, 35-39, and ≥40 years, respectively. From 2006 to 2015, the abortion rate decreased among women in all age groups. In 2015, adolescents aged <15 and 15-19 years accounted for 0.3% and 9.8% of all reported abortions, respectively, and had abortion rates of 0.5 and 6.7 abortions per 1,000 adolescents aged <15 and 15-19 years, respectively. From 2006 to 2015, the percentage of abortions accounted for by adolescents aged 15-19 years decreased 41%, and their abortion rate decreased 54%. This decrease in abortion rate was greater than the decreases for women in any older age group. In contrast to the percentage distribution of abortions and abortion rates by age, abortion ratios in 2015 and throughout the entire period of analysis were highest among adolescents and lowest among women aged 25-39 years. Abortion ratios decreased from 2006 to 2015 for women in all age groups. In 2015, almost two thirds (65.4%) of abortions were performed at ≤8 weeks' gestation, and nearly all (91.1%) were performed at ≤13 weeks' gestation. Few abortions were performed between 14 and 20 weeks' gestation (7.6%) or at ≥21 weeks' gestation (1.3%). During 2006-2015 the percentage of all abortions performed at >13 weeks' gestation remained consistently low (≤9.0%). Among abortions performed at ≤13 weeks' gestation, a shift occurred toward earlier gestational ages, with the percentage performed at ≤6 weeks' gestation increasing 11%. In 2015, 24.6% of all abortions were performed by early medical abortion (a nonsurgical abortion at ≤8 weeks' gestation), 64.3% were performed by surgical abortion at ≤13 weeks' gestation, and 8.8% were performed by surgical abortion at >13 weeks' gestation; all other methods were uncommon (≤2.2%). Among those that were eligible for early medical abortion on the basis of gestational age (i.e., performed at ≤8 weeks' gestation), 35.8% were completed by this method. In 2015, women with one or more previous live births accounted for 59.3% of abortions, and women with no previous live births accounted for 40.7%. Women with one or more previous induced abortions accounted for 43.6% of abortions, and women with no previous abortion accounted for 56.3%. Women with three or more previous births accounted for 14.2% of abortions, and women with three or more previous abortions accounted for 8.2% of abortions. Deaths of women associated with complications from abortion for 2015 are being assessed as part of CDC's Pregnancy Mortality Surveillance System. In 2014, the most recent year for which data were available, six women were identified to have died as a result of complications from legal induced abortion. Among the 49 areas that reported data every year during 2006-2015, decreases in the total number, rate, and ratio of reported abortions resulted in historic lows for the period of analysis for all three measures of abortion. The data in this report can help program planners and policymakers identify groups of women with the highest rates of abortion. Unintended pregnancy is the major contributor to induced abortion. Increasing access to and use of effective contraception can reduce unintended pregnancies and further reduce the number of abortions performed in the United States.


Charged nucleobases and their potential for RNA catalysis.
Catalysis in living cells is carried out by both proteins and RNA. Protein enzymes have been known for over 200 years, but RNA enzymes, or "ribozymes", were discovered only 30 years ago. Developing insight into RNA enzyme mechanisms is invaluable for better understanding both extant biological catalysis as well as the primitive catalysis envisioned in an early RNA-catalyzed life. Natural ribozymes include large RNAs such as the group I and II introns; small RNAs such as the hepatitis delta virus and the hairpin, hammerhead, VS, and glmS ribozymes; and the RNA portion of the ribosome and spliceosome. RNA enzymes use many of the same catalytic strategies as protein enzymes, but do so with much simpler side chains. Among these strategies are metal ion, general acid-base, and electrostatic catalysis. In this Account, we examine evidence for participation of charged nucleobases in RNA catalysis. Our overall approach is to integrate direct measurements on catalytic RNAs with thermodynamic studies on oligonucleotide model systems. The charged amino acids make critical contributions to the mechanisms of nearly all protein enzymes. Ionized nucleobases should be critical for RNA catalysis as well. Indeed, charged nucleobases have been implicated in RNA catalysis as general acid-bases and oxyanion holes. We provide an overview of ribozyme studies involving nucleobase catalysis and the complications involved in developing these mechanisms. We also consider driving forces for perturbation of the pK(a) values of the bases. Mechanisms for pK(a) values shifting toward neutrality involve electrostatic stabilization and the addition of hydrogen bonding. Both mechanisms couple protonation with RNA folding, which we treat with a thermodynamic formalism and conceptual models. Furthermore, ribozyme reaction mechanisms can be multichannel, which demonstrates the versatility of ribozymes but makes analysis of experimental data challenging. We examine advances in measuring and analyzing perturbed pK(a) values in RNA. Raman crystallography and fluorescence spectroscopy have been especially important for pK(a) measurement. These methods reveal pK(a) values for the nucleobases A or C equal to or greater than neutrality, conferring potential histidine- and lysine/arginine-like behavior on them. Structural support for ionization of the nucleobases also exists: an analysis of RNA structures in the databases conducted herein suggests that charging of the bases is neither especially uncommon nor difficult to achieve under cellular conditions. Our major conclusions are that cationic and anionic charge states of the nucleobases occur in RNA enzymes and that these states make important catalytic contributions to ribozyme activity. We conclude by considering outstanding questions and possible experimental and theoretical approaches for further advances.


DDBJ read annotation pipeline: a cloud computing-based pipeline for high-throughput analysis of next-generation sequencing data.
High-performance next-generation sequencing (NGS) technologies are advancing genomics and molecular biological research. However, the immense amount of sequence data requires computational skills and suitable hardware resources that are a challenge to molecular biologists. The DNA Data Bank of Japan (DDBJ) of the National Institute of Genetics (NIG) has initiated a cloud computing-based analytical pipeline, the DDBJ Read Annotation Pipeline (DDBJ Pipeline), for a high-throughput annotation of NGS reads. The DDBJ Pipeline offers a user-friendly graphical web interface and processes massive NGS datasets using decentralized processing by NIG supercomputers currently free of charge. The proposed pipeline consists of two analysis components: basic analysis for reference genome mapping and de novo assembly and subsequent high-level analysis of structural and functional annotations. Users may smoothly switch between the two components in the pipeline, facilitating web-based operations on a supercomputer for high-throughput data analysis. Moreover, public NGS reads of the DDBJ Sequence Read Archive located on the same supercomputer can be imported into the pipeline through the input of only an accession number. This proposed pipeline will facilitate research by utilizing unified analytical workflows applied to the NGS data. The DDBJ Pipeline is accessible at http://p.ddbj.nig.ac.jp/.


Pregnancy-induced adaptations of the central circadian clock and maternal glucocorticoids.
Maternal physiological adaptations, such as changes to the hypothalamic-pituitary-adrenal (HPA) axis, are central to pregnancy success. Circadian variation of the HPA axis is dependent on clock gene rhythms in the hypothalamus, but it is not known whether pregnancy-induced changes in maternal glucocorticoid levels are mediated via this central clock. We hypothesized that hypothalamic expression of clock genes changes across mouse pregnancy and this is linked to altered HPA activity. The anterior hypothalamus and maternal plasma were collected from C57Bl/6J mice prior to pregnancy and on days 6, 10, 14 and 18 of gestation (term=d19), across a 24-h period (0800, 1200, 1600, 2000, 0000, 0400 h). Hypothalamic expression of clock genes and Crh was determined by qPCR, plasma ACTH concentration measured by Milliplex assay and plasma corticosterone concentration by LC-MS/MS. Expression of all clock genes varied markedly across gestation, most notably at mid-gestation when levels of each gene were elevated. The pregnancy-induced increase in maternal corticosterone levels (by up to 14-fold on day 14) was not accompanied by a parallel shift in plasma ACTH (28% lower on day 14 compared with non-pregnant levels). Moreover, while circadian rhythmicity in corticosterone was maintained up to day 14 of gestation, this was effectively lost by day 18. Overall, our data show that the central circadian clock undergoes marked adaptations throughout mouse pregnancy, changes that are likely to contribute to maternal physiological adaptations. Importantly, however, neither hypothalamic clock genes nor plasma ACTH levels appear to drive the marked increase in maternal corticosterone after mid-gestation.


Computational modeling of mammalian signaling networks.
One of the most exciting developments in signal transduction research has been the proliferation of studies in which a biological discovery was initiated by computational modeling. In this study, we review the major efforts that enable such studies. First, we describe the experimental technologies that are generally used to identify the molecular components and interactions in, and dynamic behavior exhibited by, a network of interest. Next, we review the mathematical approaches that are used to model signaling network behavior. Finally, we focus on three specific instances of 'model-driven discovery': cases in which computational modeling of a signaling network has led to new insights that have been verified experimentally.


[Study of the caudate-cortical projections of the rat and cat brain by the fluorescence labeling method].
Direct caudate-cortical projections have been studied by means of the fluorescent label method. Neither in rats, nor in cats fluorescent neurons have been revealed in the caudate nucleus when primulin is injected into various cortical fields. The negative results obtained do not, however, give enough reasons to repudiate the existance of the direct caudate-cortical projections, since additional experiments performed with injection of a luminiscent dye into other brain structures demonstrate that primulin transport is not always fulfilled.


[Retrospective study of the treatment of 329 patients with prostatism syndrome and review of the literature].
New drugs for the treatment of prostatism have been developed in recent years; only surgery, however, has been demonstrated to achieve cure. Each modality of drug therapy may afford symptomatic relief in those patients in whom surgery is not indicated and a symptom-based stepwise indication could be established. 329 patients with prostatism were treated by surgery, administration of finasteride, alphablockers or plant extracts, according to the recommendations of the WHO. Patient evaluation included abdominal US. PSA determination and uroflowmetry. Patients with urodynamic or other derangements were excluded from the study. Patients with a worse quality of life and important obstructive symptoms underwent surgery. Patients with important obstructive symptoms and no complications requiring surgery (infection, urinary retention, etc.) received finasteride, which proved to be effective in these patients, although 16.6% required another type of treatment and two presented impotence. Patients treated with plant extracts had moderate prostatism; 17% of these patients required another type of treatment. Patients treated with alphablockers presented symptoms that were similar to those treated with plant extracts, but they had a more important irritative component; 24% of these patients withdrew from the study due to the side effects or inefficacy of their treatment regimen. Further studies and a longer follow-up are warranted to determine whether these new drugs can replace or effectively delay (avoiding the appearance of complications) surgery. We believe that the indications for each treatment should be established according to the different stages of prostatism. Moreover, further insight into the pathogenesis of BPH, appropriate diagnostic methods and patient selection are essential to the development of new therapeutical modalities and to achieving enhanced results.


Rat and human neutrophil N-formyl-peptide chemotactic receptors. Species difference in the glycosylation of similar 35-38 kDa polypeptide cores.
Rat and human neutrophil N-formyl-peptide chemotactic receptors were subjected to glycosidase and proteinase treatments to determine the extent and species differences of glycosylation and the carbohydrate requirement in the high-affinity ligand binding. N-Formyl-Nle-Leu-Phe-Nle-125I-Tyr-Lys was attached to rat and human neutrophils either before or after glycosidase and proteinase treatments, and the labelled receptors were solubilized after glutaraldehyde cross-linking and analysed by SDS/PAGE and autoradiography. Both the rat and human N-formyl-peptide chemotactic receptors contain only N-linked oligosaccharides, as demonstrated by their sensitivity to peptide N-glycosidase F (PNGase F) and resistance to O-glycanase treatment. The N-linked oligosaccharides seem to be of the complex type rather than the high-mannose or hybrid type and lack terminal sialic acid, as demonstrated by their resistance to endoglycosidases D and H and neuraminidase treatments. This sensitivity pattern was similar in both species, and the shift in the molecular size of the receptors to 35-38 kDa after PNGase F treatment occurred through one intermediate product, suggesting that both receptors contain a similar 35-38 kDa polypeptide core with two N-linked complex-type oligosaccharides, the heterogeneity of which is responsible for the species difference in receptor size. Papain treatment alone or followed by PNGase F produced in both species a 33-36 kDa membrane-bound fragment that was still able to bind the ligand, suggesting that the oligosaccharides are located on the approx. 2 kDa papain-cleavable polypeptide fragment of the receptors. The cleavage sites for both papain and PNGase F were hidden in occupied receptors, suggesting a conformational or topographical change in these upon ligand binding. Scatchard analyses and cross-linking experiments demonstrated that carbohydrates are not required for high-affinity ligand binding and that the 33-36 kDa membrane-bound papain fragment of both receptors contains the ligand-binding site.


Full veneers: the functional and esthetic application of bonded ceramics.
Esthetic restorative practices have used bonded ceramic facial veneers to replace or augment defective, discolored, or missing enamel. These restorations have proved to be highly esthetic and extremely durable. Sometimes it is necessary to extend the veneers onto occlusal, lingual, or proximal surfaces to cover defective tooth structure. This article will define "full veneers," present indications, show preparation designs, and present clinical cases where full veneers have been used.


A plea for intra-laboratory reference limits. Part 1. General considerations and concepts for determination.
Accurate results for quantitative procedures can be useless if the reference limits for the interpretation of laboratory results are unreliable. Recent concepts for quality management systems require that laboratories pay more attention to identification and verification of reference limits. Scientific recommendations often claim that each laboratory should determine intra-laboratory reference limits, which should be reviewed periodically. This recommendation is currently neglected by most laboratories; instead they use reference limits from external sources, despite various problems of transference. Prospective and retrospective methods either using or neglecting disease prevalences (polymodal or unimodal concepts, respectively) and applying different statistical approaches for determining reference limits have been described. The various procedures are reviewed with regard to their diagnostic sensitivity, specificity and (non-)efficiency. The present gold standard is the reference limit concept according to IFCC recommendations (a unimodal prospective approach). This concept, together with trueness-based standardization, is the most useful basis for harmonization of the decision-making process with laboratory results, despite complex problems of traceability and transference. This harmonization is at present only achieved for a limited number of analytes for which SI units and traceability can be technically realized. For the majority of measurands in laboratory medicine, much research is still required and results cannot be expected in the near future. For these measurands, a need remains for internal, efficient and simple identification of population-based reference limits. Therefore, newer retrospective concepts were developed that use large data sets from laboratory information systems to derive intra-laboratory reference limits. These approaches appear promising and should be further developed.


Sphingosine-1-phosphate enhancement of cortical actomyosin organization in cultured human Schlemm's canal endothelial cell monolayers.
Perfusion of sphingosine-1-phosphate (S1P) in whole eye organ culture models decreases outflow facility, whereas S1P promotes stress fiber formation and contractility in cultured trabecular meshwork (TM) cells. Because of S1P's known effect of increasing barrier function in endothelial cells, the authors hypothesized that Schlemm's canal (SC) cells in culture respond to S1P by increasing actomyosin organization at the cell cortex. Using primary cultures of human SC cells, the authors determined S1P activation of the GTP-binding proteins, RhoA and Rac (1,2,3). Time- and dose-dependent myosin light chain (MLC) phosphorylation in response to S1P and total expression of MLC were determined. Immunocytochemistry after S1P treatment was used to monitor filamentous actin (F-actin) and phospho-MLC organization and the localization of β-catenin, a component of adherens junctions. TM and human umbilical vein endothelial cell monolayers were used as controls. S1P (1 μM) activated RhoA and Rac after 5- and 30-minute treatments. S1P increased MLC phosphorylation with a similar time- and dose-dependent response in SC (EC(50) = 0.83 μM) compared with TM (EC(50) = 1.33 μM), though MLC expression was significantly greater in TM. In response to 1 μM S1P treatment, phospho-MLC concentrated in the SC cell periphery, coincident with cortical actin assembly and recruitment of β-catenin to the cell periphery. Results obtained in this study support the hypothesis that S1P increases actomyosin organization at the SC cell cortex and promotes intercellular junctions at the level of the inner wall of SC to increase transendothelial resistance and in part explains the S1P-induced decrease of outflow facility in organ culture.


[Cyclic changes in the population count of the taiga tick in the Stolby preserve].
A harmonic analysis was carried out of Ixodes persulcatus density dynamics over 33 years and the analysis of small mammal density and some climatic factors. It was found that the chronological changes of all the three processes contained cyclic components with the period 5.5 and 11; 3-4; 5.5 and 14-16 years, respectively. A long-term prognosis of I. persulcatus density was made based on harmonic synthesis.


Transcriptional activating activity of Smad4: roles of SMAD hetero-oligomerization and enhancement by an associating transactivator.
Smad4 plays a pivotal role in signal transduction of the transforming growth factor beta superfamily cytokines by mediating transcriptional activation of target genes. Hetero-oligomerization of Smad4 with the pathway-restricted SMAD proteins is essential for Smad4-mediated transcription. We provide evidence that SMAD hetero-oligomerization is directly required for the Smad4 C-terminal domain [Smad4(C)] to show its transcriptional transactivating activity; this requirement obtains even when Smad4(C) is recruited to promoters by heterologous DNA-binding domains and in the absence of the inhibitory Smad4 N-terminal domain. Defined mutations of GAL4 DNA-binding domain fusion of Smad4(C) that disrupt SMAD hetero-oligomerization suppressed transcriptional activation. Importantly, we found that an orphan transcriptional activator MSG1, a nuclear protein that has strong transactivating activity but apparently lacks DNA-binding activity, functionally interacted with Smad4 and enhanced transcription mediated by GAL4 DNA-binding domain-Smad4(C) and full-length Smad4. Transcriptional enhancement by MSG1 depended on transforming growth factor beta signaling and was suppressed by Smad4(C) mutations disrupting SMAD hetero-oligomerization or by the presence of Smad4 N-terminal domain. Furthermore, Smad4(C) did not show any detectable transactivating activity in yeast when fused to heterologous DNA-binding domains. These results demonstrate additional roles of SMAD hetero-oligomerization in Smad4-mediated transcriptional activation. They also suggest that the transcriptional-activating activity observed in the presence of Smad4 in mammalian cells may be derived, at least in part, from endogenously expressed separate transcriptional activators, such as MSG1.


Prediction of residual film perception of cosmetic products using an instrumental method and non-biological surfaces: The example of stickiness after skin application.
Prediction of sensory texture attributes using instrumental measurements is a very important challenge for cosmetic industry because in vivo sensory studies are expensive, time consuming and limited by the safety issue of applied products. The aim of this work is to investigate how residual sensory properties of cosmetic products can be predicted without using a panel of assessors, focusing on the residual film attribute "Stickiness". 10 cosmetic products with different galenics have been selected and evaluated in vivo using a classical sensory protocol, developed according to the Spectrum™ Descriptive Analysis method. In addition to this study, products were evaluated after their application on non-biological skin models in order to compare perceptions onto in vivo skin and artificial surfaces. Results obtained show that in vivo perceptions can be compared with the ones on artificial surfaces meaning that residual film stickiness is similar between in vivo skin and non-biological skin models. An instrumental protocol using a texture analyzer has been set up to evaluate residual film adhesiveness. This protocol has been tested and validated in vivo (r²adjusted = 0.90; RPD = 3.07) before being optimized on a selected non-biological skin model Bioskin® (Beaulax, Co. Ltd. Tokyo, Japan) owning good correlation with in vivo perceptions. Established model shows excellent predictive ability with a r²adjusted of 0.94 and a RPD of 3.38, as highlighted by the 4 steps cross-validation performed. It proves that physical stimulus responsible for cosmetic film stickiness can be instrumentally measured on both in vivo skin and artificial skin.


Identification of neutrophil subpopulations with monoclonal antibodies.
Two monoclonal antibodies have been produced by the hybridoma technique that recognize subpopulations of human neutrophils. The antibodies, termed 1B5 and 4D1, react with a mean percentage of 57% and 51% of peripheral blood granulocytes, respectively. The antigens recognized appear to be neutrophil specific in that these antibodies do not react with eosinophils, platelets, erythrocytes, monocytes, or nonadherent peripheral blood mononuclear cells. Although the neutrophil subpopulations recognized by these antibodies are nearly identical (coinclusive), the antigenic determinants recognized appear to be different. These monoclonal antibodies to neutrophil subpopulations may prove useful to studying functional heterogeneity among neutrophils as well as for investigations of normal and abnormal myeloid differentiation.


Risk Scoring System for Vancomycin-Associated Acute Kidney Injury.
Vancomycin-associated acute kidney injury (AKI) remains a major challenge for patients and clinicians. This study aimed to construct a risk scoring system for vancomycin-associated AKI. We retrospectively reviewed medical records of patients who underwent therapeutic drug monitoring for vancomycin from June 2018 to July 2019. We selected possible risk factors for AKI by univariate and multivariable logistic regression analyses and developed a scoring system for vancomycin-associated AKI. Machine learning methods were utilized to predict risk factors for the occurrence of AKI. The incidence of vancomycin-associated AKI was 31.7% among 104 patients included in this study. A bodyweight ≤60 kg (two points), a Charlson comorbidity index ≥3 (two points), a vancomycin trough serum level >15 μg/ml (one point), and concomitant use of ≥6 nephrotoxic agents (two points) were included to construct a risk scoring system based on the coefficient from the logistic regression model. The area under the receiver operating characteristic curve (AUROC) (mean, 95% confidence interval (CI)) across 10 random iterations using five-fold cross-validated multivariate logistic regression, elastic net, random forest, support vector machine (SVM)-linear kernel, and SVM-radial kernel models was 0.735 (0.638-0.833), 0.737 (0.638-0.835), 0.721 (0.610-0.833), 0.739 (0.648-0.829), and 0.733 (0.640-0.826), respectively. For total scores of 0-1, 2-3, 4-5, 6-7, the risk of vancomycin-associated AKI was 5, 25, 45, and 65%, respectively. Our scoring system can be applied to clinical settings in which several nephrotoxic agents are used along with vancomycin therapy.


Fast gap-free enumeration of conformations and sequences for protein design.
Despite significant successes in structure-based computational protein design in recent years, protein design algorithms must be improved to increase the biological accuracy of new designs. Protein design algorithms search through an exponential number of protein conformations, protein ensembles, and amino acid sequences in an attempt to find globally optimal structures with a desired biological function. To improve the biological accuracy of protein designs, it is necessary to increase both the amount of protein flexibility allowed during the search and the overall size of the design, while guaranteeing that the lowest-energy structures and sequences are found. DEE/A*-based algorithms are the most prevalent provable algorithms in the field of protein design and can provably enumerate a gap-free list of low-energy protein conformations, which is necessary for ensemble-based algorithms that predict protein binding. We present two classes of algorithmic improvements to the A* algorithm that greatly increase the efficiency of A*. First, we analyze the effect of ordering the expansion of mutable residue positions within the A* tree and present a dynamic residue ordering that reduces the number of A* nodes that must be visited during the search. Second, we propose new methods to improve the conformational bounds used to estimate the energies of partial conformations during the A* search. The residue ordering techniques and improved bounds can be combined for additional increases in A* efficiency. Our enhancements enable all A*-based methods to more fully search protein conformation space, which will ultimately improve the accuracy of complex biomedically relevant designs.


Mania following organophosphate poisoning.
Organophosphate poisoning is the most common poisoning in developing countries. Although the acute muscarinic and nicotinic side-effects of organophosphate poisoning are well known and easily recognized, but neuropsychiatric changes are rarely reported. We are reporting a case of a 33-year-old female who developed manic episode following acute organophosphate poisoning.


Allicin Regulates Energy Homeostasis through Brown Adipose Tissue.
Brown adipose tissue (BAT) is a promising potential therapeutic target for the treatment of obesity and related metabolic diseases. Allicin, a natural product in garlic, has multiple biological and pharmacological functions. However, the role of allicin in the regulation of metabolic organs, particularly BAT activation, has not been well studied. Here, we show that allicin imparts a significant effect by inhibiting body weight gain, decreasing adiposity, maintaining glucose homeostasis, improving insulin resistance, and ameliorating hepatic steatosis in obese mice. These observations strongly correlate with the activation of BAT. Notably, allicin plays a role in BAT activation, which may partly contribute to the Sirt1-PGC1α-Tfam pathway. In addition, allicin can significantly increase the succinylation levels of UCP1 in BAT by inhibiting sirt5, whereas excess allicin induces autophagy/mitophagy and mitochondrial dysfunction. Thus, our findings point to allicin as a promising therapeutic approach for the treatment of obesity and metabolic disorders.


A novel cyclic AMP response element-binding protein-1 (CREB-1) splice product may down-regulate CREB-1 activity.
In this report we identify novel spliced forms of cyclic AMP (cAMP) response element-binding protein-1 (CREB-1) mRNA. These forms contained an additional 17 nucleotide insert, which we refer to as the beta exon, located between exons 4 and 7 of the delta, and 5 and 7 of the alpha forms of CREB-1 transcript (nomenclature of Ruppert et al. 1992; EMBO Journal 11, 1503-1512). The inclusion of the beta exon led to the generation of mRNAs in which the frame of CREB-1 sequences 3' to the exon was shifted such that the encoded proteins terminate after the transactivation domain, but before the target serine for cAMP-dependent protein kinase. The beta exon-containing CREB-1 mRNAs were more abundant in tissues that respond poorly to cAMP, suggesting that the generation of beta CREB-1 mRNAs may contribute to the down-regulation of CREB-1 activity and cAMP responsiveness.


Reconstruction of large-scale gene regulatory networks using Bayesian model averaging.
Gene regulatory networks provide the systematic view of molecular interactions in a complex living system. However, constructing large-scale gene regulatory networks is one of the most challenging problems in systems biology. Also large burst sets of biological data require a proper integration technique for reliable gene regulatory network construction. Here we present a new reverse engineering approach based on Bayesian model averaging which attempts to combine all the appropriate models describing interactions among genes. This Bayesian approach with a prior based on the Gibbs distribution provides an efficient means to integrate multiple sources of biological data. In a simulation study with maximum of 2000 genes, our method shows better sensitivity than previous elastic-net and Gaussian graphical models, with a fixed specificity of 0.99. The study also shows that the proposed method outperforms the other standard methods for a DREAM dataset generated by nonlinear stochastic models. In brain tumor data analysis, three large-scale networks consisting of 4422 genes were built using the gene expression of non-tumor, low and high grade tumor mRNA expression samples, along with DNA-protein binding affinity information. We found that genes having a large variation of degree distribution among the three tumor networks are the ones that see most involved in regulatory and developmental processes, which possibly gives a novel insight concerning conventional differentially expressed gene analysis.


Serendipity in diagnostic imaging: magnetic resonance imaging of the breast.
Magnetic resonance imaging (MRI) of the breast has been proposed as a noninvasive diagnostic test for evaluation of suspicious ("index") lesions noted on mammography and/or clinical breast examination (CBE). However, women may have incidental ("serendipitous") lesions detected by MRI that are not found on mammography or CBE. To understand better whether or not biopsy procedures should be performed to evaluate serendipitous lesions, we estimated the breast cancer risk for women with this type of lesion. A decision analysis model was used to estimate the positive predictive value (i.e., the chance that a woman with a serendipitous lesion has cancer) of MRI for serendipitous lesions in women who had an abnormal mammogram and/or CBE suspicious for cancer (where a biopsy procedure is recommended). We restricted the analysis to data from women whose index lesions were noncancerous and used meta-analysis of published medical literature to determine the likelihood ratios (measures of how test results change the probability of having cancer) for MRI and the combination of CBE and mammography. The positive predictive value of MRI was calculated using the U.S. population prevalence of cancer (derived from registry data) and the likelihood ratios of the diagnostic tests. Under a wide variety of assumptions, the positive predictive value of MRI was extremely low for serendipitous lesions. For instance, assuming sensitivity and specificity values for MRI of 95.6% and 68.6%, respectively, approximately four of 1000 55- to 59-year-old women with serendipitous lesions would be expected to have cancer (positive predictive value = 0.44%, 95% confidence interval = 0.24%-0.67%). In women with a suspicious lesion discovered by mammography and/or CBE that is found to be benign, serendipitous breast lesions detected by MRI are extremely unlikely to represent invasive breast cancer. Immediate biopsy of such serendipitous lesions may, therefore, not be required.


The formation of a complex between the membrane structures of the mitochondria and DNA of nonmitochondrial origin.
The ability of decomposed rat liver mitochondria, stripped of their outer membrane (fracton C) to bind 14C-DNA of phage T4 was demonstrated. The bound phage DNA is 10-15% resistant to treatment of fraction C with DNAase I. Treatment of fraction C with sarcosyl (final concentration 0.1%) and centrifugation in a linear sucrose gradient (20-50%) permits the detection of a label in the Mt-DNA-membrane complex, isolated at 35% sucrose. Further fractionation of the membrane complex on sepharose 4 B promotes the detection of the label in "elementary particles of transcription" [1], structural units of the inner membrane, containing specific centers for binding to DNA. The centers of binding of the elementary particles of transcription possess greatest affinity for Mt-DNA; however, they are also capable of binding DNA of various origin (both the DNA of eucaryotes and that of procaryotes). The significance of fixation of DNA of nonmitochondrial origin by the membrane structures of the mitochondria is discussed.


Iron homeostasis: a new job for macrophages in adipose tissue?
Elevated serum ferritin and increased cellular iron concentrations are risk factors for diabetes; however, the etiology of this association is unclear. Metabolic tissues such as pancreas, liver, and adipose tissue (AT), as well as the immune cells resident in these tissues, may be involved. Recent studies demonstrate that the polarization status of macrophages has important relevance to their iron-handling capabilities. Furthermore, a subset of macrophages in AT have elevated iron concentrations and a gene expression profile indicative of iron handling, a capacity diminished in obesity. Because iron overload in adipocytes increases systemic insulin resistance, iron handling by AT macrophages may have relevance not only to adipocyte iron stores but also to local and systemic insulin sensitivity.


Survival and prognostic factors in systemic sclerosis-associated pulmonary hypertension: a systematic review and meta-analysis.
Pulmonary hypertension (PH) is a frequent and life-limiting complication of systemic sclerosis (SSc). However, data on survival rates and their evolution over time, as well as prognostic factors in SSc complicated by PH, are still conflicting. The aim of this study was to conduct a systematic review and meta-analysis of cohort studies to assess pooled survival and prognostic factors for survival in patients with SSc-associated PH. For this systematic review and meta-analysis, we searched the Medline and EMBase databases (January 1960 to January 2012). All cohort studies in which survival and/or prognostic factors for SSc-associated PH were reported were included in the analysis. We calculated the pooled survival rates and analyzed their evolution over time and identified prognostic factors for survival. Twenty-two studies were included, representing a total of 2,244 patients with SSc-associated PH. The pooled 1-, 2-, and 3-year survival rates were 81% (95% confidence interval [95% CI] 79-84%), 64% (95% CI 59-69%), and 52% (95% CI 47-58%), respectively. Meta-regression did not reveal a significant change in survival over time, while baseline hemodynamic measures of PH severity were significantly correlated with survival. In patients with SSc complicated by pulmonary arterial hypertension (PAH), age, male sex, diffusing capacity for carbon monoxide (DLCO), pericardial effusion, and the parameters classically associated with the severity of idiopathic PAH, including the 6-minute walk distance, mean pulmonary artery pressure, cardiac index, and right atrial pressure, were significant prognostic factors. DLCO and pericardial effusion were the only prognostic factors in patients with interstitial lung disease-related PH. Our meta-analysis revealed a poor pooled 3-year survival rate of 52% in patients with SSc-associated PH. Baseline hemodynamic measures of PAH severity, but not the period of time during which patients were included in the studies, correlated significantly with survival in patients with SSc-associated PAH. All of the prognostic factors typically observed in idiopathic PAH, including the 6-minute walk distance and right atrial pressure, were also prognostic factors in SSc-associated PAH.


Manipulation of domain wall mobility by oxygen vacancy ordering in multiferroic YMnO3.
The mobility of the ferroelectric domain phases and the local conductivity of ferroelectric domain walls in multiferroic YMnO3 crystals grown in air and reduced atmosphere were studied by piezoresponse force microscopy (PFM), tip-enhanced Raman spectroscopy (TERS) and conductive atomic force microscopy (CAFM). Oxygen vacancies were found to reduce the strength of 4d-2p (Y(3+)-O(2-)) hybridization and structural trimerization distortion, leading to the disappearance of the six wedge-shaped ferroelectric domain phases in oxygen deficient YMnO3-δ crystals. We observed anisotropic domain wall motion such that the wedge-shaped domain configuration joined at one point could be changed to the stripe domain configuration by applying high electric fields in oxygen deficient YMnO3-δ single crystals. The local conductivity of the domain walls increased significantly in poled YMnO3-δ single crystals. The straight conductive domain walls in YMnO3-δ, instead of the twisted insulating ones in the stoichiometric crystal, are induced by the ordered oxygen vacancies which are verified by TERS measurements.


The Roles of Physical Activity and Inflammation in Mortality, Cognition, and Depressive Symptoms Among Older Mexican Americans.
A higher level of physical activity (PA) is associated with decreased risk of mortality, dementia, and depression, yet the mechanisms involved are not well understood, and little evidence exists for Mexican Americans. With data from the Sacramento Area Latino Study on Aging (1998-2007), we used Cox proportional hazards regression to separately evaluate associations of baseline PA level with mortality, dementia/cognitive impairment without dementia (CIND), and depressive symptoms, and we estimated the mediating effects of inflammatory markers in additive hazard models. A low level of PA (<35 metabolic equivalent of task-hours/week) was associated with increased mortality (hazard ratio (HR) = 1.50, 95% confidence interval (CI): 1.20, 1.88), dementia/CIND (HR = 1.37, 95% CI: 0.96, 1.96), and depressive symptoms (HR = 1.23, 95% CI: 1.00, 1.52). A low PA level added 512 (95% CI: -34, 1,058) cases of dementia/CIND per 100,000 person-years at risk (direct effect), while, through a mediating path, interleukin 6 (IL-6) added another 49 (95% CI: 5, 94) cases, or 9% of the total effect. For mortality, 8%-10% of the PA total effect was mediated through IL-6, tumor necrosis factor α (TNF-α), or TNF-α receptors. None of the inflammatory markers mediated the association between PA and depressive symptoms. Our results suggest that antiinflammation (especially as assessed by IL-6 and TNF-α levels) may partly explain how PA protects against dementia/CIND and mortality.


Selective antinociceptive effects of a combination of the N-methyl-D-aspartate receptor peptide antagonist [Ser(1)]histogranin and morphine in rat models of pain.
Numerous rather than a few analgesic endogenous neuropeptides are likely to work in concert in vivo in ameliorating pain. Identification of effective neuropeptide combinations would also facilitate the development of gene or cell-based analgesics. In this study, opioid peptides endomorphin-1 (EM-1) and endomorphin-2 (EM-2) and the peptide histogranin analogue [Ser(1)]histogranin (SHG), which possess activity as an N-methyl-d-aspartate (NMDA) receptor antagonist, were intrathecally (i.t.) injected alone and in combination in rat models of acute and persistent pain. None of the peptides when injected alone altered hind paw responses of uninjured rats to acute noxious stimulation. EM-1 and EM-2 showed divergent efficacies in the persistent pain models. For example, EM-1 injected alone was antinociceptive in rats with neuropathic pain, whereas EM-2 demonstrated no efficacy. Demonstration of synergism was also divergent across the models. For example, while SHG combined with EM-1 did not alter the efficacy of EM-1 in rats with neuropathic pain, SHG significantly increased the efficacy of EM-1 in the formalin test. By contrast, the potency and efficacy of the peptides alone and combinations were much less than those of the reference analgesic morphine. Furthermore, morphine combined with the clinically used NMDA receptor antagonist ketamine showed synergism across a broad range of pain states. While the current set of neuropeptides could serve as a basis for analgesic therapeutics, there could be other neuropeptides with greater efficacy and potency and broader therapeutic application.