Subunit structures of different electrophoretic forms of nucleosomes.
We have reported previously that five different electrophoretic forms of mononucleosomes (MI to MV) are produced upon treatment of mammalian chromatin with micrococcal nuclease. We show here that each of these mononucleosome classes possesses internal heterogeneity due to the presence of a variety of minor protein species. Defined subsets of mononucleosome classes MII to MV have been reconstituted by reassociating stripped nucleosomes with histone H1 and non-histone protein HMG-17. This procedure leads to the generation of the same five major electrophoretic forms of mononucleosomes found in native chromatin. From the results of one- and two-dimensional electrophoretic analyses on reconstituted samples, it is concluded that different mononucleosome classes possess the following subunit structures: MI, core histone octamer (8-mer); MII, 8-mer plus one copy of HMG-17; MIIIA, 8-mer plus one copy of histone H1; MIIIB, 8-mer plus two copies of HMG-17; MIV, 8-mer plus one copy each of histone H1 and HMG-17; and MV, 8-mer plus one copy of histone H1 and two copies of HMG-17. Equal numbers of HMG-14 molecules can substitute for HMG-17 and generate the same nucleosome components. Thus, mononucleosomes possess independent binding sites for at least 1 histone H1 molecule and 2 nonhistone chromosomal protein molecules. We show further that reassociated HMG-17 molecules can exhibit a rapid interchange between binding sites, even under conditions of low ionic strength.


Evidence for a brief period of enhanced oxygen susceptibility in the rat model of oxygen-induced retinopathy.
Findings in a previous study have shown that the retina of newborn rats exposed to hyperoxia during the first days of life sustain permanent functional (as determined with the rod ERG) and structural (as determined with histology) damage that appears to be determined by the level of retinal maturity reached at the time of oxygen exposure-the retinas of rat pups being more susceptible to hyperoxic shock during the second week of life than during the first week. Given that the cone ERG has been shown to mature later than the rod ERG, the purpose of the present study was to examine whether cone responses also demonstrates a similar maturational susceptibility to postnatal hyperoxia. Also examined was whether the oscillatory potentials (OPs) were affected by postnatal hyperoxia. Newborn rats were exposed to hyperoxia during selected postnatal day intervals either initiated at birth (early-onset exposure) or at a later postnatal age (late-onset exposure). Photopic and scotopic (mixed cone-rod) electroretinograms were recorded at 30 days. Data analysis reveals that photopic and scotopic responses (b-wave and OPs) demonstrated a similar maturational susceptibility to postnatal hyperoxia, in which exposure regimens initiated during the second week of life were most detrimental to retinal function. The results also revealed a temporal window of enhanced oxygen susceptibility at approximately postnatal day 10. The duration of this window was longer when estimated with the scotopic responses, but the extent of the functional damage was more pronounced when estimated with the photopic signals. Finally, compared with the b-wave, the OPs, especially the short-latency OPs, were proportionally more affected. The results suggest that cone function is significantly more susceptible to postnatal hyperoxia than rod function, and the OPs appear to be the most susceptible ERG components, thus suggesting a differential susceptibility to oxygen toxicity of the different retinal components. However, despite a clear demonstration of its existence, the exact nature of the temporal window of enhanced oxygen susceptibility as well as a possible equivalence in other animal models of oxygen induced retinopathy, including the human form (retinopathy of prematurity), remains to be determined.


Evidence for the role of infectious disease in species extinction and endangerment.
Infectious disease is listed among the top five causes of global species extinctions. However, the majority of available data supporting this contention is largely anecdotal. We used the IUCN Red List of Threatened and Endangered Species and literature indexed in the ISI Web of Science to assess the role of infectious disease in global species loss. Infectious disease was listed as a contributing factor in <4% of species extinctions known to have occurred since 1500 (833 plants and animals) and as contributing to a species' status as critically endangered in <8% of cases (2,852 critically endangered plants and animals). Although infectious diseases appear to play a minor role in global species loss, our findings underscore two important limitations in the available evidence: uncertainty surrounding the threats to species survival and a temporal bias in the data. Several initiatives could help overcome these obstacles, including rigorous scientific tests to determine which infectious diseases present a significant threat at the species level, recognition of the limitations associated with the lack of baseline data for the role of infectious disease in species extinctions, combining data with theory to discern the circumstances under which infectious disease is most likely to serve as an agent of extinction, and improving surveillance programs for the detection of infectious disease. An evidence-based understanding of the role of infectious disease in species extinction and endangerment will help prioritize conservation initiatives and protect global biodiversity.


[The age of Gutenberg is over: a consideration of medical education--past, present and future].
Education is the basis for reliable medical care and medical progress. Our medical knowledge has increased more in the past 50 years than in the 500 years before. The spatial and human resource capacity of our universities cannot cope with the existing academic structures and needs. Part of the problem can be solved by "blended learning", that is a combination of traditional teaching methods (frontal lectures, courses, bedside teaching) with supplementary web-based e-learning. In addition to conveying a sound basic knowledge, the ability to cope with modern media and prepare for lifelong learning must also be taught. Out of the large number of e-learning platforms for undergraduate students offered in the internet, we present the program DOIT (Dermatology Online with Interactive Technology; http://www.swisdom.org) and the program Dermokrates (http://www.Dermokrates.com) of the German, Austrian and Swiss Dermatological Societies for postgraduate Continuing Medical Education (CME). The biggest obstacle in the implementation of new developments is the stubborn adherence to traditional structures.


A Cuban perspective on management of persistent vegetative state.
The Cuban Group for Study of Disorders of Consciousness is developing several research protocols to search for possible preservation of residual brain and autonomic functions in cases of persistent vegetative and minimally conscious states. We present examples showing the importance of 3D anatomic reconstruction of brain structures and MRI tractography for assessing white matter connectivity. We also present results of use of proton magnetic resonance spectroscopy technique to follow up cognitive recovery in persistent vegetative state patients transitioning to minimally conscious state. We have demonstrated recognition of a mother's voice with emotional content after zolpidem administration, indicating high-level residual linguistic processing and brain activation despite the patient's apparent inability to communicate. Hence we differ with current thinking that, by definition, subjects in persistent vegetative state are isolated from the outside world and cannot experience pain and suffering. We also consider "vegetative state" a pejorative term that should be replaced. KEYWORDS Persistent vegetative state, minimally conscious state, consciousness disorders, magnetic resonance imaging, electroencephalography, heart rate variability.


p53 expression and clinical outcome in prostate cancer.
Abnormally high levels of expression of p53 protein are found in many human cancers. In most cases increased expression is associated with point mutations in one allele of the p53 gene and loss of the other allele. Accumulation of the protein product can be detected by immunohistochemistry. p53 protein expression in 68 men with prostate cancer, followed up for at least 8 years or until death, was assessed by immunohistochemistry. The aim of the study was to determine the association between p53 protein expression, cell cycling and clinical outcome. Nine (13%) of 68 tumours stained positively for p53; all 9 tumours were category T3 or T4. p53 positive tumours had a significantly greater Gleason score than p53 negative tumours. Eight of the 9 p53 positive tumours had > 10% cells in G2 + mitosis, compared with 61% of p53 negative tumours. All 17 patients with p53 positive tumours available for follow-up progressed clinically, compared with 28 of 38 patients (74%) with p53 negative tumours. The median time to progression was 12 months in p53 positive tumours and 24 months in p53 negative tumours. Median survival in p53 positive tumours was 40 months, compared with 76 months in p53 negative tumours. This study demonstrates that overexpression of p53 in a small population of prostate cancers is associated with a poor prognosis in terms of progression and survival.


Chromosomal phylogeny and evolution of the African mole-rats (Bathyergidae).
The subterranean African mole-rats (Family Bathyergidae) show considerable variation in their diploid numbers, but there is limited understanding of the events that shaped the extant karyotypes. Here we investigate chromosomal evolution in specimens representative of six genera and an outgroup species, the cane rat Thryonomys swinderianus, using flow-sorted painting probes isolated from the naked mole-rat, Heterocephalus glaber (2n = 60). A chromosomal phylogeny based on the cladistic analysis of adjacent syntenies detected by cross-species chromosome painting was not consistent with that obtained using DNA sequences due, in large part, to the conserved nature of the Bathyergus, Georychus and Cryptomys karyotypes. In marked contrast, the Fukomys and Heliophobius species showed extensive chromosome reshuffling, permitting their analysis by a computational approach that has conventionally been employed in comparative genomic studies for retrieving phylogenetic information based on DNA sequence or gene order data. Using the multiple genome rearrangements (MGR) algorithm and chromosomal rearrangement data detected among F. damarensis, F. darlingi, F. mechowi and the sister taxa B. janetta and Heliophobius argenteocinereus, cytogenetic support for the monophyly of Fukomys and a sister association for F. darlingi + F. damarensis was retrieved, mirroring the published sequence-based topology. We show that F. damarensis, a lineage adapted to arid and climatically unpredictable environments in Southern Africa, is characterized by a large number of fissions the fixation of which has probably been favoured by environmental factors and/or its particular eusocial structure.


The emergence of pattern in embryogenesis: regulation of beta-catenin localization during early sea urchin development.
The accumulation of beta-catenin in the nuclei of blastomeres at one pole of the early embryo is a highly conserved and essential feature of animal development. In the sea urchin, beta-catenin accumulates in the nuclei of vegetal blastomeres during early cleavage and activates gene regulatory networks that drive mesoderm and endoderm formation. Measurements of beta-catenin half-life in vivo have demonstrated a gradient in stability along the animal-vegetal axis. Dishevelled (Dsh), a protein that regulates beta-catenin turnover, is localized in the vegetal cortex, where it has an essential role in stabilizing beta-catenin and activating endomesodermal gene networks. Two motifs of Dsh are required for targeting to the vegetal cortex. Overexpression of Dsh in animal blastomeres does not alter their fate, which suggests that a localized activator of Dsh may be missing in these cells. Wnt signaling may be localized in the early sea urchin embryo, as it is in Xenopus, but findings point to possible differences in the initial polarizing signal in amphibians and echinoderms. Further studies will be required to determine the extent to which mechanisms that control beta-catenin nuclearization in early embryogenesis have been conserved during animal evolution.


In Vitro Adhesion, Invasion, and Transcytosis of Streptococcus pneumoniae with Host Cells.
Physical interactions of bacteria with host cells are often a principal aspect of bacterial pathogenesis. In the case of Streptococcus pneumoniae (Spn), which does not produce a secreted toxin, adhesion to and/or invasion of host cells is necessary for colonization of the nasopharynx and subsequently to cause opportunistic disease in its human host. Knowledge of how pneumococci interact with host cells thereby helps to explain its biology and may identify potential targets for intervention. One of the simplest, yet powerful, assays that can be leveraged to dissect the molecular basis of this vital host-pathogen interaction is the in vitro adhesion and invasion assay. Among many key results, this assay has been used to discover the bacterial and host determinants involved in bacterial attachment, identify host signaling networks required for uptake of the bacteria into an endosome, and the characterization of the intracellular trafficking machinery that is subverted by Spn during development of bacteremia and meningitis. These assays have also been used to characterize the epithelial, endothelial, and/or immune cell response to these bacteria, and to learn how pneumococci disperse from an established biofilm to a planktonic phenotype to colonize another niche and/or transmit. Herein, we will review this protocol, highlighting how simple changes in the bacterial strain or host cell line can elucidate the underlying molecular mechanisms for Spn virulence.


"Women enjoy punishment": attitudes and experiences of gender-based violence among PHC nurses in rural South Africa.
Violence against women is pervasive in South Africa where, as in many other countries, cultural values and norms serve to condone and reinforce abusive practices against women. Primary health care nurses, who are widely distributed throughout the rural areas, may appear to be an ideal network for addressing this issue in resource-poor settings. However, based on a qualitative and quantitative study of a class of 38 PHC nurses, this paper emphasises that the nurses are women and men first--and as such, experience the same cultural values, and indeed, similar or higher levels of violence, as the clients they are expected to counsel and treat. Current models for encouraging nurses and other health care workers to detect and address gender-based violence have evolved largely in the context of developed countries, and have focused primarily on acquiring the knowledge, skills and attitudes necessary to engage this issue in the health care setting. Yet, as this paper suggests, there is an urgent need to understand and address the lived experiences of the nurses, and the duality of their roles as professionals and as community members, before promoting the training of nurses as an effective strategy for dealing with gender-based violence. One such training model was piloted and assessed in this study. The intervention used partnership with a domestic violence NGO to initially focus on dealing with the attitudes and experiences of the nurses as individuals, and to begin a process of self-awareness and sensitisation. Only then did the intervention turn to their roles as professional nurses. Clearly, there is a need for further research to explore these issues in more depth and to inform the development of appropriate training strategies for health care workers, particularly in developing countries. Moreover, such research may well have implications for the design and implementation of training interventions aimed at raising awareness and capacity within other sectors such as the welfare, police and judicial systems.


Susceptibility of BALB/c mice carrying various DBA/2 genes to development of Friend murine leukemia virus-induced erythroleukemia.
Using a series of BALB/c mice congenic for various DBA/2 genes, we were able to establish that DBA/2 mice carry a gene on chromosome 5, at or near the Rmcfr locus, that plays a major role in resistance to early erythroleukemia induced by injection of Friend murine leukemia virus (F-MuLV) into newborn mice. The fact that this gene controls the replication of mink cell focus-inducing (MCF) viruses strengthens the case for these viruses playing a crucial role in the development of erythroleukemia, since failure to replicate MCF viruses results in resistance to early erythroleukemia. The expression of the Rmcfr gene is correlated with the constitutive expression of an MCF virus-related envelope glycoprotein that apparently blocks the receptor for MCF viruses, preventing their spread. Thus, the Rmcfr gene is either a structural gene for this unique protein, which can block the receptor for MCF viruses, or is a regulatory gene that controls expression of such a structural gene. Although the Rmcfr gene is clearly involved in resistance to the early erythroleukemia induced by F-MuLV, it appears to have no effect on the late myeloid, lymphoid or erythroid diseases that appear in DBA/2 and other strains of mice after injection of F-MuLV, consistent with data indicating that replication of MCF viruses is not required for the development of these late diseases. Our studies with congenic and backcross mice also indicate that, in addition to the Rmcfr gene, other genes of DBA/2 origin may contribute to resistance to F-MuLV-induced early erythroleukemia by mechanisms other than blocking the replication of MCF viruses.


Treatment of a Class II subdivision based on occlusal plane control: a clinical case.
It is important to understand the morphologic features of malocclusion to construct a functional occlusion. The features of Class II subdivision malocclusions with mandibular lateral deviation make them especially difficult to correct. This article describes a Class II subdivision clinical case, treated at the first stage with the straightwire technique, which permitted alignment and leveling for 12 months. At the second stage, an occlusal plane modification and mandible repositioning with multiloop edgewise archwires (MEAW) was used for 6 months. Different MEAW activation on either side and occlusal provisional composite restorations (prematurity) in the maxillary left first and second molars became essential to increase the vertical dimension in the affected side, thereby allowing the left premolars to extrude into a Class I relationship. Short vertical elastics in the anterior area (for Class III) on the right side and on the left side (Class II) also applied different orthodontic forces. This procedure made it possible to finish the case with stable sagittal and vertical occlusion relationships between both arches.


Yoga Nidra as a complementary treatment of anxiety and depressive symptoms in patients with menstrual disorder.
Emotional insecurity, stress, depressive or/and anxiety symptoms are common with variable severity among patients with menstrual disorder. Yogic relaxation therapy (Yoga Nidra) leads to conscious and subconscious recognition of these underlying psychological factors and helps releasing of suppressed conflicts. To evaluate the effect of Yoga Nidra on anxiety and depressive symptoms in patients with menstrual disorders. Subjects were recruited from the Department of Obstetrics and Gynecology, C.S.M. Medical University (erstwhile KGMU), Lucknow Uttar Pradesh, India. The subjects were randomly divided in to two groups: Intervention group (with yogic intervention) and control group (without yogic intervention). Assessments of all subjects were carried out by administering Hamilton anxiety scale (HAM-A) and Hamilton rating scale for depression (HAM-D) at baseline and after six months. The mean age with S.D of the intervention group was 27.67 ± 7.85 years, and for control group was 26.58 ± 6.87 years (among completed intervention group nn = 65 and control group nn = 61). There was significant reduction of scores in HAM-A (P<0.003) and HAM-D (P<0.02) respectively in subjects with mild to moderate anxiety and depressive symptoms after six months of yoga therapy (Yoga Nidra) in intervention group in comparison to control group. The patients with mild to moderate anxiety and depressive symptoms improve significantly with 'Yoga Nidra' intervention. There is no significant improvement in the patients with severe anxiety and depressive symptoms.


Multiple GTP-binding proteins participate in clathrin-coated vesicle-mediated endocytosis.
We have examined the effects of various agonists and antagonists of GTP-binding proteins on receptor-mediated endocytosis in vitro. Stage-specific assays which distinguish coated pit assembly, invagination, and coat vesicle budding have been used to demonstrate requirements for GTP-binding protein(s) in each of these events. Coated pit invagination and coated vesicle budding are both stimulated by addition of GTP and inhibited by GDP beta S. Although coated pit invagination is resistant to GTP gamma S, A1F4-, and mastoparan, late events involved in coated vesicle budding are inhibited by these antagonists of G protein function. Earlier events involved in coated pit assembly are also inhibited by GTP gamma S, A1F4-, and mastoparan. These results demonstrate that multiple GTP-binding proteins, including heterotrimeric G proteins, participate at discrete stages in receptor-mediated endocytosis via clathrin-coated pits.


Condom use during most recent vaginal intercourse event among a probability sample of adults in the United States.
Correct and consistent condom use remains the most effective way to reduce sexually transmissible infection/HIV transmission during sex and is a highly effective contraceptive method. Understanding correlates of condom use is vital to public health programs. To explore sociodemographic, event characteristics, and experiential correlates of condom use at last penile-vaginal intercourse (PVI). Survey data were collected from a nationally representative probability sample of adults in the United States as part of the National Survey of Sexual Health and Behavior. Condom use/non-use at most recent PVI was the main outcome. Logistic regression analyses predicted condom use from sociodemographic variables (i.e., age, education, race/ethnicity, sexual orientation, health status, type of relationship with sexual partner) and event characteristics (i.e., location of sexual encounter, prior intercourse experience with partner, whether partner had other sex partners in the 6 months prior to sex with the participant; other contraceptive use, alcohol use, marijuana use, and for men, erection medication use). Logistic regression analyses examined evaluations of the sexual aspects of the experience (pleasure, arousal, erection/lubrication difficulty, participant orgasm, partner orgasm) in light of condom use. Condom-protected PVI was significantly greater among younger people, blacks and Hispanics, and those having PVI with a nonrelationship partner. Statistically adjusting for these differences, condom use was significantly associated with fewer previous intercourse experiences with the partner and not using other forms of contraception. The sexual aspects of experience were evaluated similarly regardless of whether or not a condom was used. Public health programs among youths and minorities may underlie higher condom use rates among these groups. Condom use may be further improved by continuing such programs and also expanding outreach to older persons and whites, suggesting prolonging use as relationships develop, and highlighting that condom use does not necessarily interfere with the sexual experience.


[Drug evaluation of huperzine A in the treatment of senile memory disorders].
Huperzine A is an alkaloid which was first isolated from Huperzia serrata (Thumb) Trev by Zhejiang Academy of Medical Sciences and Shanghai Institute of Materia Medica, Chinese Academy of Sciences. It exhibits a significant anticholinesterase activity and has been used on myasthenia gravis patients. The therapeutic effects were studied by random, match and double-blind method on 56 patients of multi-infarct dementia or senile dementia and 104 patients of senile and presenile simple memory disorders. The curative effects were evaluated by Wechsler memory scale. The im dose for multi-infarct dementia was 0.05 mg bid for 4 wk, whereas that for senile and presenile simple memory disorders was 0.03 mg bid for 2 wk. Saline was used on control group. The result showed that the curative effect of huperzine A was significant. Only a few patients felt slight dizziness and this did not affect the therapeutic effects.


Intravenous low redox potential saline attenuates FeCl3-induced vascular dysfunction via downregulation of endothelial H2O2, CX3CL1, intercellular adhesion molecule-1, and p53 expression.
Exaggerated reactive oxygen species (ROS) may contribute to vascular injury by the enhancement of CX3CL1, intercellular adhesion molecule-1 (ICAM-1), and pro-apoptotic p53 expression. Reduced water with safely antioxidant activity may protect vascular tissue against oxidative injury. We established reduced water (RW) by using a modified magnesium alloy and evaluated the effects of an RW-made culture medium on TNF-α-induced endothelial damage in vitro and intravenous RW-made saline (0.9%NaCl) infusion on FeCl(3)-induced arterial injury in rats in vivo. Several oxidative stresses were evaluated by using a chemiluminescence analyzer, Western blot, and immunohistochemistry. We found that the established RW, RW-culture medium, and RW saline displayed a lower redox potential (<-150 mV) and efficient H(2)O(2) scavenging activity compared with distilled-water-made solutions. The RW-culture medium significantly depressed TNF-α-enhanced endothelial H(2)O(2) production; improved CX3CL1, ICAM-1, and p53 expression; and inhibited activated monocyte adhesion to endothelial cells as well as to the CX3CL1 or the ICAM-1 coated plate when compared with the distilled-water-culture medium. In the in vivo study, the time required for FeCl(3)-induced occlusion in the urethane anesthetized rat's carotid and femoral arteries was significantly extended by intravenous RW saline infusion compared with distilled-water saline. FeCl(3) stimulation significantly enhanced vascular NADPH oxidase activity, ROS production, as well as CX3CL1, ICAM-1, p53, 3-nitrotyrosine, and 4-hydroxynonenal expression in the damaged arteries. Intravenous RW saline significantly reduced all the FeCl(3)-enhanced oxidative parameters when compared with intravenous distilled-water-saline infusion. We conclude that the RW-culture medium and saline made from magnesium alloy confer cardiovascular protection by the antioxidant capability.


Sequence determination of a mildly virulent strain (CU-2) of Gallid herpesvirus type 2 using 454 pyrosequencing.
The complete DNA sequence of the mildly virulent Gallid herpesvirus type 2 strain CU-2 was determined and consists of 176,922 bp with an overall gene organization typical of class E herpesviruses. Phylogenetically, this strain partitions in its own branch between the virulent strains RB-1B, Md11, and Md5, and the vaccine strain CVI988. Overall, the genome of CU-2 is more similar to that of CVI988, with identically sized unique short regions of 11,651 bp. As in CVI988, an insertion of 177 bp was identified in the overlapping genes encoding the Meq, RLORF6, and 23 kDa proteins within the repeat long region of the genome. A total of 15 single nucleotide polymorphisms (SNPs) common to both CU-2 and CVI988, and not occurring in virulent strains, were identified in the genes encoding UL29, UL45, UL50, UL52, LORF10, RLORF14a, RLORF12, Meq(RLORF7), 23kDa, ICP4, US3, and two hypothetical proteins MDV071.4 and MDV076.4. Each gene encoding UL29 and Meq contained two SNPs. Only one major open reading frame (ORF) encoding UL41, the virus host shutoff (VHS) ribonuclease, was disrupted in the CU-2 genome. An additional cytosine after the 25 codon is predicted to produce a truncated protein of 97 aa. Since GaHV-2 mutants lacking UL41 have been reported to retain their virulence, other factors are likely responsible for the low virulence of CU-2. It is largely suspected that SNPs in common with CVI988 along with the insertions in the Meq loci are responsible for its phenotype. Conversely, we identified 43 nonsynonymous mutations (within 23 genes) that may contribute to the virulence of CU-2. These SNPs are shared exclusively with all sequenced virulent strains (Md5, Md11, and RB-1B) and not present within the CVI988 genome. Although most occur in proteins of unknown function, a significant percentage is in proteins involved in virion assembly.


Safety and efficacy of a pharmaco-invasive reperfusion strategy in rural ST-elevation myocardial infarction patients with expected delays due to long-distance transfers.
To determine the safety and efficacy of a pharmaco-invasive reperfusion strategy utilizing half-dose fibrinolysis combined with transfer for immediate percutaneous coronary intervention (PCI) in ST-elevation myocardial infarction (STEMI) patients presenting to remote rural hospitals. Primary PCI is preferred for STEMI if performed in a timely manner. However, <20% of STEMI patients transferred for PCI in the USA have door-to-balloon times <2 h. Prospective data from the Level 1 MI programme were analysed. All STEMI patients presenting to the Minneapolis Heart Institute or 31 referral hospitals received aspirin, clopidogrel, and unfractionated heparin (UFH) at the presenting hospital and those presenting to hospitals ≥60 miles away also received half-dose fibrinolytic with transfer for immediate PCI. From April 2003 through December 2009, we enrolled 2634 consecutive STEMI patients in the Level 1 MI database including 660 transferred from remote hospitals utilizing pharmaco-invasive therapy and 600 patients who presented directly to the PCI centre. There were no significant differences in 30-day mortality (5.5 vs. 5.6%; P= 0.94), stroke (1.1 vs. 1.3%; P= 0.66) or major bleeding (1.5 vs. 1.8%; P= 0.65), or re-infarction/ischaemia (1.2 vs. 2.5%; P= 0.088) in patients receiving a pharmaco-invasive strategy compared with patients presenting directly to the PCI centre, despite a significantly longer door-to-balloon time. Within a regional STEMI system of care, half-dose fibrinolysis combined with immediate transfer for PCI may be a safe and effective option for STEMI patients with expected delays due to long-distance transfer.


A Gro/TLE-NuRD corepressor complex facilitates Tbx20-dependent transcriptional repression.
The cardiac transcription factor Tbx20 has a critical role in the proper morphogenetic development of the vertebrate heart, and its misregulation has been implicated in human congenital heart disease. Although it is established that Tbx20 exerts its function in the embryonic heart through positive and negative regulation of distinct gene programs, it is unclear how Tbx20 mediates proper transcriptional regulation of its target genes. Here, using a combinatorial proteomic and bioinformatic approach, we present the first characterization of Tbx20 transcriptional protein complexes. We have systematically investigated Tbx20 protein-protein interactions by immunoaffinity purification of tagged Tbx20 followed by proteomic analysis using GeLC-MS/MS, gene ontology classification, and functional network analysis. We demonstrate that Tbx20 is associated with a chromatin remodeling network composed of TLE/Groucho corepressors, members of the Nucleosome Remodeling and Deacetylase (NuRD) complex, the chromatin remodeling ATPases RUVBL1/RUVBL2, and the T-box repressor Tbx18. We determined that the interaction with TLE corepressors is mediated via an eh1 binding motif in Tbx20. Moreover, we demonstrated that ablation of this motif results in a failure to properly assemble the repression network and disrupts Tbx20 function in vivo. Importantly, we validated Tbx20-TLE interactions in the mouse embryonic heart, and identified developmental genes regulated by Tbx20-TLE binding, thereby confirming a primary role for a Tbx20-TLE repressor complex in embryonic heart development. Together, these studies suggest a model in which Tbx20 associates with a Gro/TLE-NuRD repressor complex to prevent inappropriate gene activation within the forming heart.


Effects of serotonin depletion on punishment processing in the orbitofrontal and anterior cingulate cortices of healthy women.
Diminished synthesis of the neurotransmitter serotonin (5-HT) has been linked to disrupted impulse control in aversive contexts. However, the neural correlates underlying a serotonergic modulation of female impulsivity remain unclear. The present study investigated punishment-induced inhibition in healthy young women. Eighteen healthy female subjects (aged 20-31) participated in a double-blinded, counterbalanced, placebo-controlled, within subjects, repeated measures study. They were assessed on two randomly assigned occasions that were controlled for menstrual cycle phase. In a randomized order, one day, acute tryptophan depletion (ATD) was used to reduce 5-HT synthesis in the brain. On the other day, participants received a tryptophan-balanced amino acid load (BAL) as a control condition. Three hours after administration of ATD/BAL, neural activity was recorded during a modified Go/No-Go task implementing reward or punishment processes using functional magnetic resonance imaging (fMRI). Neural activation during No-Go trials in punishment conditions after BAL versus ATD administration correlated positively with the magnitude of central 5-HT depletion in the ventral and subgenual anterior cingulate cortices (ACC). Furthermore, neural activation in the medial orbitofrontal cortex (mOFC) and the dorsal ACC correlated positively with trait impulsivity. The results indicate reduced neural sensitivity to punishment after short-term depletion of 5-HT in brain areas related to emotion regulation (subgenual ACC) increasing with depletion magnitude and in brain areas related to appraisal and expression of emotions (mOFC and dorsal ACC), increasing with trait impulsivity. This suggests a serotonergic modulation of neural circuits related to emotion regulation, impulsive behavior, and punishment processing in females.


Detecting Tripartite Steering via Quantum Entanglement.
Einstein-Podolsky-Rosen steering is a kind of powerful nonlocal quantum resource in quantum information processing such as quantum cryptography and quantum communication. Many criteria have been proposed in the past few years to detect steerability, both analytically and numerically, for bipartite quantum systems. We propose effective criteria for tripartite steerability and genuine tripartite steerability of three-qubit quantum states by establishing connections between the tripartite steerability (resp. genuine tripartite steerability) and the tripartite entanglement (resp. genuine tripartite entanglement) of certain corresponding quantum states. From these connections, tripartite steerability and genuine tripartite steerability can be detected without using any steering inequalities. The "complex cost" of determining tripartite steering and genuine tripartite steering can be reduced by detecting the entanglement of the newly constructed states in the experiment. Detailed examples are given to illustrate the power of our criteria in detecting the (genuine) tripartite steerability of tripartite states.


Prevalence and Correlates of Perceived Infertility in Ghana.
Perceived infertility is an understudied phenomenon in low- and middle-income countries, where biomedical infertility can have severe consequences, particularly for women. We conducted a nationally representative survey of Ghanaian women, estimated the prevalence of and reasons for perceived infertility, and assessed factors associated with higher levels of perceived infertility using a partial proportional odds model. Among 4,070 women, 13 percent believed they were "very likely" to have difficulty getting pregnant when they wanted to, 21 percent believed this was "somewhat likely," and 66 percent believed this was "not at all likely." Reasons for perceived infertility varied by whether the respondent was currently seeking pregnancy. In multivariable analysis, several factors were associated with higher levels of perceived infertility, while unexpectedly, women who reported ever using contraception were less likely to report perceived infertility. Acknowledging the need to address infertility globally and understanding the role of perceived infertility are important components in supporting people's ability to decide whether and when to have children.


ASXL1 mutations are infrequent in young patients with primary acute myeloid leukemia and their detection has a limited role in therapeutic risk stratification.
ASXL1 mutations are recurrent in acute myeloid leukemia (AML), but it is unclear whether ASXL1 genotype might influence patient management. We analyzed frequency and impact in younger (15-59 years) and older (≥ 60 years) patients with primary or secondary disease. Overall, 9% had truncating mutations. Incidence was significantly lower in younger patients with primary than with secondary disease (4%, 12%; p = 0.03). In older patients it did not differ significantly (11%, 15%; p = 0.5). In univariate analysis, ASXL1-mutated patients had a worse outcome (5-year relapse 83% vs. 56%, p = 0.01; overall survival [OS] 6% vs. 22%, p = 0.02). However in multivariate analysis, ASXL1 mutations had no prognostic significance (for OS, p = 0.3), because age was a major confounding factor. The low incidence of mutations in younger patients with primary disease and the lack of significance in multivariate analysis indicate that there is a limited role for screening at diagnosis for ASXL1 mutations for the purpose of prognostic stratification.


Restrictions for reimbursement of interferon-free direct-acting antiviral drugs for HCV infection in Europe.
All-oral direct-acting antiviral drugs (DAAs) for hepatitis C virus, which have response rates of 95% or more, represent a major clinical advance. However, the high list price of DAAs has led many governments to restrict their reimbursement. We reviewed the availability of, and national criteria for, interferon-free DAA reimbursement among countries in the European Union and European Economic Area, and Switzerland. Reimbursement documentation was reviewed between Nov 18, 2016, and Aug 1, 2017. Primary outcomes were fibrosis stage, drug or alcohol use, prescriber type, and HIV co-infection restrictions. Among the 35 European countries and jurisdictions included, the most commonly reimbursed DAA was ombitasvir, paritaprevir, and ritonavir, with dasabuvir, and with or without ribavirin (33 [94%] countries and jurisdictions). 16 (46%) countries and jurisdictions required patients to have fibrosis at stage F2 or higher, 29 (83%) had no listed restrictions based on drug or alcohol use, 33 (94%) required a specialist prescriber, and 34 (97%) had no additional restrictions for people co-infected with HIV and hepatitis C virus. These findings have implications for meeting WHO targets, with evidence of some countries not following the 2016 hepatitis C virus treatment guidelines by the European Association for the Study of Liver.


A guide to the understanding and use of tricyclic antidepressants in the overall management of fibromyalgia and other chronic pain syndromes.
The purpose of this review is to present relatively detailed information on the characteristics of tricyclic antidepressants, mainly amitriptyline hydrochloride and doxepin hydrochloride, for use as an integral part of the safe and effective management of fibromyalgia and, to a lesser extent, other chronic pain syndromes. Data sources include MEDLINE searches in English, relevant reference books and textbooks, my personal database and library, as well as personal clinical experience. I discuss these data with regard to the pharmacologic characteristics, mechanisms of action, adverse effects, and precautions involved with the use of tricyclic antidepressants. Additional information is given on drug selection and dosage titration. Much emphasis is placed on the fact that while tricyclic antidepressants play a major role in the management of fibromyalgia and other chronic pain syndromes, lifestyle alterations (eg. physical reconditioning and exercise), as well as behavior modification, are also vital to a successful outcome in management.


Burkholderia cepacia complex in cystic fibrosis and non-cystic fibrosis patients: identification of a cluster of epidemic lineages.
This study was performed in order to compare Burkholderia cepacia complex strains from cystic fibrosis (CF) and non-CF patients at the genomovar, genetic and epidemiological levels. A total of 92 B. cepacia respiratory tract isolates were obtained from patients attending the following CF centres: Catania and Palermo, Sicily; Gualdo Tadino, Central Italy, and Milan, Northern Italy. A total of 23 B. cepacia isolates were obtained from blood, surgical wound, and intravenous catheter sources of patients without CF, hospitalized in Catania and Varese, Northern Italy. Genomovar status identification, clonality and genetic relatedness determination, antibiotic susceptibility pattern determination and electron microscopy were performed. Transmission of infection was shown in both CF and non-CF patients by identifying clonality of responsible strains. In total 13 clones were involved in cross-transmission episodes. No outbreak was described involving both CF and non-CF patients. The present study indicates the existence of a distinct cluster of strains responsible for epidemics in CF and non-CF patients, based on their genetic relatedness, distinct from strains associated with no or negligible transmissibility. This result suggests that transmissibility is not only associated with a specific genomovar in CF patients, but also with a group of genetically related lineages in CF and non-CF patients. A key role is shown for both segregation measures and careful surveillance of infection, based on selective culture, molecular identification and epidemiological characterization of individual isolates.


Disruption of hyaluronan synthase-2 abrogates normal cardiac morphogenesis and hyaluronan-mediated transformation of epithelium to mesenchyme.
We identified hyaluronan synthase-2 (Has2) as a likely source of hyaluronan (HA) during embryonic development, and we used gene targeting to study its function in vivo. Has2(-/-) embryos lack HA, exhibit severe cardiac and vascular abnormalities, and die during midgestation (E9.5-10). Heart explants from Has2(-/-) embryos lack the characteristic transformation of cardiac endothelial cells into mesenchyme, an essential developmental event that depends on receptor-mediated intracellular signaling. This defect is reproduced by expression of a dominant-negative Ras in wild-type heart explants, and is reversed in Has2(-/-) explants by gene rescue, by administering exogenous HA, or by expressing activated Ras. Conversely, transformation in Has2(-/-) explants mediated by exogenous HA is inhibited by dominant-negative Ras. Collectively, our results demonstrate the importance of HA in mammalian embryogenesis and the pivotal role of Has2 during mammalian development. They also reveal a previously unrecognized pathway for cell migration and invasion that is HA-dependent and involves Ras activation.


2-Amino-pyridinium 2-meth-oxy-carbonyl-4,6-dinitro-phenolate.
In the title mol-ecular salt, C(5)H(7)N(2) (+)·C(8)H(5)N(2)O(7) (-), the 2-amino-pyridinium cation is essentially planar, with a maximium deviation of 0.015 (1) Å, while the 2-meth-oxy-carbonyl-4,6-dinitro-phenolate anion is slightly twisted away from planarity, with a maximium deviation of 0.187 (1) Å. Deprotonation of the hy-droxy O atom was observed. The cation and anion are connected by four bifurcated N-H⋯(O,O) hydrogen bonds, forming a mol-ecular proton-transfer adduct. The dihedral angle between the pyridinium ring in the cation and the benzene ring in the anion is 3.65 (6)°. Every adduct connects to six neighboring adducts by N-H⋯O and C-H⋯O hydrogen bonds, yielding extended layers parallel to the bc plane. There is a weak π-π inter-action between the benzene rings of two neighboring anions; the inter-planar spacing and the centroid-centroid separation are 3.309 (1) and 3.69 (1) Å, respectively.


The BiP protein and the endoplasmic reticulum of Schizosaccharomyces pombe: fate of the nuclear envelope during cell division.
A polyclonal antibody was raised to the C-terminal region of fission yeast BiP. The use of this antibody for immunoprecipitation, western blotting and immunofluorescence has confirmed and extended the observations made previously with an epitope-tagged BiP molecule. A fraction of BiP protein is glycosylated in Schizosaccharomyces pombe cells. Pulse-chase experiments showed that this modification occurs rapidly upon synthesis and that the extent of glycosylation does not then change with time. BiP protein is induced by elevated temperatures and by treatment with tunicamycin. The antibody cross-reacts with proteins of similar molecular weight in the yeasts Kluyveromyces lactis and Schizosaccharomyces japonicus. Immunofluorescence of BiP has been used to follow the behaviour of the ER and in particular the nuclear envelope through the cell cycle.


Tenascin-C Promotes Tumor Cell Migration and Metastasis through Integrin α9β1-Mediated YAP Inhibition.
Tenascin-C is an extracellular matrix molecule that drives progression of many types of human cancer, but the basis for its actions remains obscure. In this study, we describe a cell-autonomous signaling mechanism explaining how tenascin-C promotes cancer cell migration in the tumor microenvironment. In a murine xenograft model of advanced human osteosarcoma, tenascin-C and its receptor integrin α9β1 were determined to be essential for lung metastasis of tumor cells. We determined that activation of this pathway also reduced tumor cell-autonomous expression of target genes for the transcription factor YAP. In clinical specimens, a genetic signature comprising four YAP target genes represents prognostic impact. Taken together, our results illuminate how tumor cell deposition of tenascin-C in the tumor microenvironment promotes invasive migration and metastatic progression.Significance: These results illuminate how the extracellular matrix glycoprotein tenascin-C in the tumor microenvironment promotes invasive migration and metastatic progression by employing integrin α9β1, abolishing actin stress fiber formation, inhibiting YAP and its target gene expression, with potential implications for cancer prognosis and therapy. Cancer Res; 78(4); 950-61. ©2017 AACR.


Abcc6 deficiency in mice leads to altered ABC transporter gene expression in metabolic active tissues.
ATP-binding cassette (ABC) transporters are involved in a huge range of physiological processes. Mutations in the ABCC6 gene cause pseudoxanthoma elasticum, a metabolic disease with progressive soft tissue calcification. The aim of the present study was to analyze gene expression levels of selected ABC transporters associated with cholesterol homeostasis in metabolic active tissues, such as the liver, kidney and white adipose tissue (WAT) of Abcc6-/- mice from an early and late disease stage (six-month-old and 12-month-old mice). The strongest regulation of ABC transporter genes was observed in the liver tissue of six-month-old Abcc6-/- mice. Here, we found a significant increase of mRNA expression levels of phospholipid, bile salt and cholesterol/sterol transporters Abcb1b, Abcb11, Abcg1, Abcg5 and Abcg8. Abcd2 mRNA expression was increased by 3.2-fold in the liver tissue. We observed strong upregulation of Abca3 and Abca1 mRNA expression up to 3.3-fold in kidney and WAT, and a 2-fold increase of Abca9 mRNA in the WAT of six-month-old Abcc6 knockout mice. Gene expression levels of Abcb1b and Abcg1 remained increased in the liver tissue after an age-related disease progression, while we observed lower mRNA expression of Abca3 and Abca9 in the kidney and WAT of 12-month-old Abcc6-/- mice. These data support previous findings that Abcc6 deficiency leads to an altered gene expression of other ABC transporters depending on the status of disease progression. The increased expression of fatty acid, bile salt and cholesterol/sterol transporters may be linked to an altered cholesterol and lipoprotein metabolism due to a loss of Abcc6 function.


The epigenome as a potential mediator of cancer and disease prevention in prenatal development.
Epigenetic events establish a particular gene expression signature for each cell type during differentiation and fertilization. Disruption of these epigenetic programs in response to environmental stimuli during prenatal exposure dysregulates the fetal epigenome, potentially impacting susceptibility to disease later in life (the fetal basis of adult disease). Maternal dietary modifications during gestation and lactation play a pivotal role in the period of fetal (re)programming. Recently, many studies have demonstrated the impact of maternal nutrition on the fetal epigenome. This review discusses the complex interplay among various environmental factors and epigenetic mechanisms that have been found to affect offspring in human and animal models. Further, it summarizes the impact of various dietary phytochemicals capable of modulating the epigenome with regard to diverse human cancers and childhood cancer, specifically those with potential environmental etiology through maternal consumption during pregnancy and lactation. Other dietary agents that are still untested as to their effectiveness in transplacental studies are also discussed. The recent developments discussed herein enhance current understanding of how chemopreventive agents act and their potential to impact the prenatal epigenome; they may also aid efforts to identify dietary interventions that can be beneficial in treating and preventing disease.


Ways of dying.
How do we die? Is it an event or a process? Does everyone die in the same way or are there different ways of dying? Even with humankind's claims to gigantic strides in knowledge, death still remains one of the great mysteries for the living. And that makes it the subject of profound and perennial philosophical and religious enquiry. Modern medical science, however, had no option but to engage with it in its bodily form and try to define the precise nature of the process of death. Things were rather easy when death was equal to stoppage of the heart. But inevitably, the world of medicine with its keen sense of observation and constant yearning for scientific reasoning, recognised, somewhere in the middle of the last century, that the brains of a set of individuals hooked onto support systems in intensive care units were dying first, inevitably followed by the heart after some time. The tremendous progress in the science of resuscitation, organ support and intensive care while saving many lives resulted in an increasing number of such individuals, who were soon termed "brain dead" but whose hearts were still beating. The damage to the brain was irreversible and complete and inevitably, the heart had to follow within hours, or sometimes days.


Comparative study of ultrasound, 131I-19-iodocholesterol scintigraphy, and aortography in localising adrenal lesions.
Twenty-seven consecutive patients with endocrinological disease necessitating adrenal surgery underwent blind preoperative investigation with ultrasound, renal aortography, and adrenocortical scintigraphy for an adrenal lesion. Nine patients had pituitary-dependent Cushing's syndrome, five had pituitary-independent Cushing's syndrome, four had an adrenocortical androgenic excess, and nine had a preoperative diagnosis of phaeochromocytoma. The predictive value of preoperative ultrasound was 100% for a positive finding and 79% for a negative result. Preoperative aortography had a predictive value of 83% for a positive finding and 64% for a negative result; and the predictive value of adrenocortical scintigraphy was 100% for a positive finding and 85% for a negative finding. In localising biochemically suspected adrenal lesions ultrasound should be the first choice, since it is rapid, noninvasive, cheap, and reasonably accurate. Adrenocortical scintigraphy has a similar diagnostic value, especially in Cushing's syndrome, but it is time consuming. Nevertheless, it may be preferable for diagnosing small glucocorticoid-secreting adenomas. Aortography should be reserved for cases with inconclusive diagnoses and suspected extra-adrenal phaeochromocytomas.


Contributions of incidence and case fatality to mortality from bladder cancer in the south Thames Regions.
The aim was to assess the individual contributions of incidence and case fatality to variations in bladder cancer mortality between districts in the South Thames Regions. The standardised mortality ratios for bladder cancer were calculated for the health districts in the South Thames Regions using data from the Thames Cancer Registry. The results were compared with the standardised registration ratios, used as a measurement of incidence, and survival hazard function, used as a measurement of case fatality. As one determinant of case fatality is the severity at presentation, mortality was also compared with the standardised proportion of cases with advanced disease at diagnosis. The study took place in the 28 health districts in South East and South West Thames regional health authorities. Subjects were 3271 cases of bladder cancer aged 15-74 years resident in one of the two South Thames regional health authorities at diagnosis and registered between 1982 and 1985. The standardised mortality ratios for the different districts varied from 62 to 139. The standardised registration ratios, the survival hazard functions adjusted for age and stage of disease at diagnosis, and the proportion of cases with severe disease at diagnosis were all independently and significantly related to the standardised mortality ratios in each district. Mortality from bladder cancer was significantly related to measurements of incidence, case fatality, and severity at presentation. These relations have implications for the health services. The incidence of disease may be modified through preventative measures, the case fatality through improved quality of care, and severity at presentation possibly through prompt management of patients with haematuria. Further studies are needed to investigate why these factors have high values in some districts.


Metastatic lung cancer in the age of targeted therapy: improving long-term survival.
Epidermal growth factor receptor (EGFR) mutations are the most frequent targetable genetic abnormality observed in non-small cell lung cancer (NSCLC). More than a decade after EGFR mutations were shown to predict sensitivity to EGFR-tyrosine kinase inhibitors (EGFR-TKI), retrospective cohort studies are now identifying and characterizing 5-year survivors. While these studies indicate subsets of patients achieving long-term survival, there is paucity of data pertaining to the long-term survival benefits of these targeted therapies at a population level. Improving access to molecular testing and treatment are key to maximizing the survival benefits at a population level.


Toxic shock syndrome in children: epidemiology, pathogenesis, and management.
Toxic shock syndrome (TSS) is an acute, toxin-mediated illness, like endotoxic shock, and is characterized by fever, rash, hypotension, multiorgan involvement, and desquamation. TSS reflects the most severe form of the disease caused by Staphylococcus aureus and Streptococcus pyogenes. A case definition for staphylococcal TSS was well established in the early 1980s and helped in defining the epidemiology. Since the late 1980s, a resurgence of highly invasive streptococcal infections, including a toxic shock-like syndrome, was noted worldwide and a consensus case definition for streptococcal TSS was subsequently proposed in 1993. Both TSS and the toxic shock-like syndrome occur at a lower incidence in children than in adults. Changes in the manufacturing and use of tampons led to a decline in staphylococcal TSS over the past decade, while the incidence of nonmenstrual staphylococcal TSS increased. Nonmenstrual TSS and menstrual TSS are now reported with almost equal frequency. The incidence of streptococcal TSS remains constant after its resurgence, but varies with geographic location. Streptococcal TSS occurs most commonly following varicella or during the use of NSAIDs. Sites of infection in streptococcal TSS are much deeper than in staphylococcal TSS, such as infection caused by blunt trauma, and necrotizing fasciitis. Bacteremia is more common in streptococcal TSS than in staphylococcal TSS. Mortality associated with streptococcal TSS is 5-10% in children, much lower than in adults (30-80%), and is 3-5% for staphylococcal TSS in children.TSS is thought to be a superantigen-mediated disease. Toxins produced by staphylococci and streptococci act as superantigens that can activate the immune system by bypassing the usual antigen-mediated immune-response sequence. The host-pathogen interaction, virulence factors, and the absence or presence of host immunity determines the epidemiology, clinical syndrome, and outcome. Early recognition of this disease is important, because the clinical course is fulminant and the outcome depends on the prompt institution of therapy. Management of a child with TSS includes hemodynamic stabilization and appropriate antimicrobial therapy to eradicate the bacteria. Supportive therapy, aggressive fluid resuscitation, and vasopressors remain the main elements. An adjuvant therapeutic strategy may include agents that can block superantigens, such as intravenous immunoglobulin that contains superantigen neutralizing antibodies.


Sensory and multisensory responses in the newborn monkey superior colliculus.
Superior colliculus (SC) neurons have the ability to synthesize information from different sensory modalities, resulting in enhancements (or depressions) of their activity. This physiological capacity is, in turn, closely tied to changes in overt attentive and orientation responses. The present study shows that, in contrast to more altricial species, many deep layer SC neurons in the rhesus monkey are multisensory at birth. Such neurons can respond to stimuli from different sensory modalities, and all convergence patterns seen in the adult are represented. Nevertheless, these neurons cannot yet synthesize their multisensory inputs. Rather, they respond to combinations of cross-modal stimuli much like they respond to their individual modality-specific components. This immature property of multisensory neurons is in contrast to many of the surprisingly sophisticated modality-specific response properties of these neurons and of their modality-specific neighbors. Thus, although deep SC neurons in the newborn have longer latencies and larger receptive fields than their adult counterparts, they are already highly active and are distributed in the typical adult admixture of visual, auditory, somatosensory, and multisensory neurons. Furthermore, the receptive fields of these neurons are already ordered into well organized topographic maps, and the different receptive fields of the same multisensory neurons show a good degree of cross-modal spatial register. These data, coupled with those from cat, suggest that the capacity to synthesize multisensory information does not simply appear in SC neurons at a prescribed maturational stage but rather develops only after substantial experience with cross-modal cues.


Cryptosporidium is more closely related to the gregarines than to coccidia as shown by phylogenetic analysis of apicomplexan parasites inferred using small-subunit ribosomal RNA gene sequences.
The phylogenetic placement of gregarine parasites (Apicomplexa: Gregarinasina) within the Apicomplexa was derived by comparison of small-subunit ribosomal RNA gene sequences. Gregarine sequences were obtained from Gregarina niphandrodes Clopton, Percival, and Janovy, 1991, and Monocystis agilis Stein, 1848 (Eugregarinorida Léger 1900), as well as from Ophriocystis elektroscirrha McLaughlin and Myers, 1970 (Neogregarinorida Grassé 1953). The sequences were aligned with several other gregarine and apicomplexan sequences from GenBank and the resulting data matrix analyzed by parsimony and maximum-likelihood methods. The gregarines form a monophyletic clade that is a sister group to Cryptosporidium spp. The gregarine/ Cryptosporidium clade is separate from the other major apicomplexan clade containing the coccidia, adeleids, piroplasms, and haemosporinids. The trees indicate that the genus Cryptosporidium has a closer phylogenetic affinity with the gregarines than with the coccidia. These results do not support the present classification of the Cryptosporidiidae in the suborder Eimerioirina Léger, 1911.


Quantitative analysis of miRNA expression in seven human foetal and adult organs.
miRNAs have been found to repress gene expression at posttranscriptional level in cells. Studies have shown that expression of miRNAs is tissue-specific and developmental-stage-specific. The mechanism behind this could be explained by miRNA pathways. In this study, totally 54 miRNAs were analysed in 7 matched human foetal and adult organs (brain, colon, heart, kidney, liver, lung and spleen) using real-time PCR. Quantitative analysis showed that a big proportion of the 54 miRNAs have higher general expression in the organs of the foetal period than the adult period, with the exception of the heart. The miRNA gene promoter methylation level in the adult stages was higher than in the foetal stages. Moreover, there is a high general expression level of several miRNAs in both stages of brain, kidney, liver, lung and spleen, but not seen in colon and heart. Our results indicate that the miRNAs may play a bigger role in the foetal stage than the adult stage of brain, colon, kidney, liver, lung and spleen. The majority of the miRNAs analysed may play an important role in the growth and development of brain, kidney, liver, lung and spleen. However, a minority of the miRNAs may be functional in colon and heart.


Developmental changes in cardiac myocyte calcium regulation.
Developmental changes in the contributions of transsarcolemmal Ca2+ influx and Ca2+ release from intracellular storage sites to myocardial contraction were evaluated in isolated ventricular myocytes from neonatal (aged 1-7 days) and adult (aged 8-10 weeks) New Zealand White rabbits. Contractions ceased in one beat when extracellular Ca2+ was decreased from 1mM to micromolar levels using a rapid perfusion technique. On reperfusion with 1 mM Ca2+, recovery of control contraction amplitude occurred after significantly fewer beats in neonatal myocytes compared with adult myocytes, and after 1 minute compared with 5 minutes of reduced Ca2+. After 15 minutes of perfusion with either 1 or 10 microM ryanodine, contraction amplitude decreased in both age groups, but the decrease was significantly greater in adults than in neonates. These experiments indicate that isolated ventricular myocytes may be used in the study of developmental changes in intracellular Ca2+ regulation. Results suggest that cardiac contraction in neonates is relatively more dependent on transsarcolemmal Ca2+ influx. Furthermore, although Ca2+ release from intracellular storage sites is present in both neonates and adults, its role in cardiac contraction is more significant in adults.


Excision repair of nitrogen mustard-DNA adducts in Saccharomyces cerevisiae.
The bifunctional alkylating anticancer drug nitrogen mustard forms a variety of DNA lesions, including monoadducts and intrastrand and interstrand crosslinks. Although it is known that nucleotide excision repair (NER) is important in processing these adducts, the role of the other principal excision repair pathway, base excision repair (BER) is less well defined. Using isogenic Saccharomyces cerevisiae strains disrupted for a variety of NER and BER genes we have examined the relative importance of the two pathways in the repair of nitrogen mustard adducts. As expected, NER defective cells (rad4 and rad14 strains) are extremely sensitive to the drug. One of the BER mutants, a 3-methyladenine glycosylase defective (mag1) strain also shows significant hypersensitivity. Using a rad4/mag1 double mutant it is shown that the two excision repair pathways are epistatic to each other for nitrogen mustard sensitivity. Furthermore, both rad14 and mag1 disruptants show elevated levels of nitrogen mustard-induced forward mutation. Measurements of repair rates of nitrogen mustard N-alkylpurine adducts in the highly transcribed RPB2 gene demonstrate defects in the processing of mono-adducts in rad4, rad14 and mag1 strains. However, there are differences in the kinetics of adduct removal in the NER mutants compared to the mag1 strain. In the mag1 strain significant repair occurs within 1 h with evidence of enhanced repair on the transcribed strand. Adducts however accumulate at later times in this strain. In contrast, in the NER mutants repair is only evident at times greater than 1 h. In a mag1/rad4 double mutant damage accumulates with no evidence of repair. Comparison of the rates of repair in this gene with those in a different genomic region indicate that the contributions of NER and BER to the repair of nitrogen mustard adducts may not be the same genome wide.


Determinants of integrated health care development: chains of care in Sweden.
Local health care in Sweden is an emerging form of integrated care, linked together by chains of care. Experiences show, however, that the development of chains of care is making slow progress. In order to study the factors behind this development, an embedded multiple-case study design was chosen. The study compared six health authorities in Sweden, three with successful and three with unsuccessful chain of care development. Three major determinants of integrated health care development were identified: professional dedication, legitimacy and confidence. In more detail, space for prime movers and trust between participants were crucial success factors, while top-down approaches targeting at the same time a change of management systems were negative for the development of chains of care. Resistance from the body of physicians was a serious obstacle to such a development. Local health care depends on developed chains of care, but it seems that health care managers do not have the management systems necessary to run these clinical networks, mainly due to a lack of acceptance from the medical profession. This is an impossible situation in the long run, since the number of chains of care is likely to increase as a result of the emerging local health care.


Epo receptor signaling in macrophages alters the splenic niche to promote erythroid differentiation.
Anemic stress induces stress erythropoiesis, which rapidly generates new erythrocytes to restore tissue oxygenation. Stress erythropoiesis is best understood in mice where it is extramedullary and occurs primarily in the spleen. However, both human and mouse stress erythropoiesis use signals and progenitor cells that are distinct from steady-state erythropoiesis. Immature stress erythroid progenitors (SEPs) are derived from short-term hematopoietic stem cells. Although the SEPs are capable of self-renewal, they are erythroid restricted. Inflammation and anemic stress induce the rapid proliferation of SEPs, but they do not differentiate until serum erythropoietin (Epo) levels increase. Here we show that rather than directly regulating SEPs, Epo promotes this transition from proliferation to differentiation by acting on macrophages in the splenic niche. During the proliferative stage, macrophages produce canonical Wnt ligands that promote proliferation and inhibit differentiation. Epo/Stat5-dependent signaling induces the production of bioactive lipid mediators in macrophages. Increased production of prostaglandin J2 (PGJ2) activates peroxisome proliferator-activated receptor γ (PPARγ)-dependent repression of Wnt expression, whereas increased production of prostaglandin E2 (PGE2) promotes the differentiation of SEPs.


Performance of Cpred/Cobs concentration ratios as a metric reflecting adherence to antidepressant drug therapy.
Nonadherence is very common among subjects undergoing pharmacotherapy for schizophrenia and depression. This study aimed to evaluate the performance of the ratio of the nonlinear mixed effects pharmacokinetic model predicted concentration to observed drug concentration (ratio of population predicted to observed concentration (Cpred/Cobs) and ratio of individual predicted to observed concentration (Cipred/Cobs) as a measure of erratic drug exposure, driven primarily by variable execution of the dosage regimen and unknown true dosage history. Modeling and simulation approaches in conjunction with dosage history information from the Medication Event Monitoring System (MEMS, provided by the "Depression: The search for treatment relevant phenotypes" study), was applied to evaluate the consistency of exposure via simulation studies with scenarios representing a long half-life drug (escitalopram). Adherence rates were calculated based on the percentage of the prescribed doses actually taken correctly during the treatment window of interest. The association between Cpred/Cobs, Cipred/Cobs ratio, and adherence rate was evaluated under various assumptions of known dosing history. Simulations for those scenarios representing a known dosing history were generated from historical MEMS data. Simulations of a long half-life drug exhibited a trend for overprediction of concentrations in patients with a low percentage of doses taken and underprediction of concentrations in patients taking more than their prescribed number of doses. Overall, the ratios did not predict adherence well, except when the true adherence rates were extremely high (greater than 100% of prescribed doses) or extremely low (complete nonadherence). In general, the Cipred/Cobs ratio was a better predictor of adherence rate than the Cpred/Cobs ratio. Correct predictions of extreme (high, low) 7-day adherence rates using Cipred/Cobs were 73.8% and 64.0%. This simulation study demonstrated the limitations of the Cpred/obs and Cipred/obs ratios as metrics for actual dosage intake history, and identified that use of MEMS dosing history monitoring combined with sparse pharmacokinetic sampling is a more reliable approach.


Non-classical actions of testosterone: an update.
Androgens are known to exert their effects via genomic signalling, which involves intracellular androgen receptors that modulate gene expression on steroid binding. Whereas non-classical estrogen effects are well established, it is only recently that non-classical, rapid, membrane-initiated testosterone actions have received attention. Non-classical effects of testosterone have now been demonstrated convincingly in several tissues, in particular in the reproductive, cardiovascular, immune and musculoskeletal systems. There is evidence for the participation of the classical intracellular androgen receptor and for involvement of novel, membrane-associated androgen receptors in the non-classical actions of testosterone. Here we discuss evidence for rapid testosterone actions, which have clinical implications in fertility, cardiovascular disease and the treatment of prostate cancer.


Identification of small scale biochemical networks based on general type system perturbations.
New technologies enable acquisition of large data-sets containing genomic, proteomic and metabolic information that describe the state of a cell. These data-sets call for systematic methods enabling relevant information about the inner workings of the cell to be extracted. One important issue at hand is the understanding of the functional interactions between genes, proteins and metabolites. We here present a method for identifying the dynamic interactions between biochemical components within the cell, in the vicinity of a steady-state. Key features of the proposed method are that it can deal with data obtained under perturbations of any system parameter, not only concentrations of specific components, and that the direct effect of the perturbations does not need to be known. This is important as concentration perturbations are often difficult to perform in biochemical systems and the specific effects of general type perturbations are usually highly uncertain, or unknown. The basis of the method is a linear least-squares estimation, using time-series measurements of concentrations and expression profiles, in which system states and parameter perturbations are estimated simultaneously. An important side-effect of also employing estimation of the parameter perturbations is that knowledge of the system's steady-state concentrations, or activities, is not required and that deviations from steady-state prior to the perturbation can be dealt with. Time derivatives are computed using a zero-order hold discretization, shown to yield significant improvements over the widely used Euler approximation. We also show how network interactions with dynamics that are too fast to be captured within the available sampling time can be determined and excluded from the network identification. Known and unknown moiety conservation relationships can be processed in the same manner. The method requires that the number of samples equals at least the number of network components and, hence, is at present restricted to relatively small-scale networks. We demonstrate herein the performance of the method on two small-scale in silico genetic networks.


Iron as a bound secondary electron donor in modified bacterial reaction centers.
The binding and oxidation of ferrous iron were studied in wild-type reaction centers and in mutants that have been modified to be both highly oxidizing and able to bind manganese [Thielges et al. (2005) Biochemistry 44, 7389-7394]. After illumination of wild-type reaction centers, steady-state optical spectroscopy showed that the oxidized bacteriochlorophyll dimer, P+, could oxidize iron but only as a second-order reaction at iron concentrations above 100 microM. In the modified reaction centers, P+ was reduced by iron in the presence of sodium bicarbonate with dissociation constants of approximately 1 microM for two mutants with different metal-binding sites. Transient optical spectroscopy showed that P+ was rapidly reduced with first-order rates of 170 and 275 s-1 for the two mutants. The dependence of the amplitude of this rate on the iron concentration yielded a dissociation constant of approximately 1 microM for both mutants, in agreement with the steady-state determination. The oxidation of bound iron by P+ was confirmed by the observation of a light-induced EPR signal centered at g values of 2.2 and 4.3 and attributed to high-spin Fe3+. Bicarbonate was required at pH 7 for low dissociation constants for both iron and manganese binding. The similarity between iron and manganese binding in these mutants provides insight into general properties of metal-binding sites in proteins.


Temporal pathways of change in two randomized controlled trials for depression and harmful drinking in Goa, India.
The current study explored the temporal pathways of change within two treatments, the Healthy Activity Program (HAP) for depression and the Counselling for Alcohol Problems (CAP) Program for harmful drinking. The study took place in the context of two parallel randomized controlled trials in Goa, India. N = 50 random participants who met a priori criteria were selected from each treatment trial and examined for potential direct and mediational pathways. In HAP, we examined the predictive roles of therapy quality and patient-reported activation, assessing whether activation mediated the effects of therapy quality on depression (Patient Health Questionnaire-9) outcomes. In CAP, we examined the predictive roles of therapy quality and patient change- and counter-change-talk, assessing whether change- or counter-change-talk mediated the effects of therapy quality on daily alcohol consumption. In HAP, therapy quality (both general and treatment-specific skills) was associated with patient activation; patient activation but not therapy quality significantly predicted depression outcomes, and patient activation mediated the effects of higher general skills on subsequent clinical outcomes [a × b = -2.555, 95% confidence interval (CI) -5.811 to -0.142]. In CAP, higher treatment-specific skills, but not general skills, were directly associated with drinking outcomes, and reduced levels of counter-change talk both independently predicted, and mediated the effects of higher general skills on, reduced alcohol consumption (a × b = -24.515, 95% CI -41.190 to -11.060). Change talk did not predict alcohol consumption and was not correlated with counter-change talk. These findings suggest that therapy quality in early sessions operated through increased patient activation and reduced counter-change talk to reduce depression and harmful drinking respectively.


Neurophysiological responses in the brain tissues of rainbow trout (Oncorhynchus mykiss) treated with bio-pesticide.
The aim of this study was to investigate neurophysiological responses in rainbow trout brain tissue exposed to natural/botanical pesticides. Fish were exposed to botanical and synthetic pesticides over a 21-day period. At the end of the treatment period, oxidative DNA damage (indicated by 8-OHdG (8-hydroxy-2'-deoxyguanosine), AChE activity (acetylcholinesterase) and transcriptional parameters (gpx (glutathione peroxidase), sod (superoxide dismutase), cat (catalase), HSP70 (heat shock protein 70) and CYP1A (cytochromes P450)) was investigated in control and application groups. Our results indicated that brain AChE activities decreased very significantly in fish treated with both insecticide types when compared with control (p < 0.05). 8-OHdG activity increased in a dose/time-dependent situation in the brain tissues of Oncorhynchus mykiss (p < 0.05). In addition, with regards to gene expression, gpx sod and, cat expressions were down-regulated, whereas CYP1A and HSP70 gene expression were up-regulated in fish treated with both insecticides when compared to the control group (p < 0.05). The data for this study suggests that bio-pesticides can cause neurophysiological changes in fish brain tissue.


Hyperglycemia cooperates with Tet2 heterozygosity to induce leukemia driven by proinflammatory cytokine-induced lncRNA Morrbid.
Diabetes mellitus (DM) is a risk factor for cancer. The role of DM-induced hyperglycemic (HG) stress in blood cancer is poorly understood. Epidemiologic studies show that individuals with DM are more likely to have a higher rate of mutations in genes found in pre-leukemic hematopoietic stem and progenitor cells (pre-LHSPCs) including TET2. TET2-mutant pre-LHSPCs require additional hits to evolve into full-blown leukemia and/or an aggressive myeloproliferative neoplasm (MPN). Intrinsic mutations have been shown to cooperate with Tet2 to promote leukemic transformation. However, the extrinsic factors are poorly understood. Using a mouse model carrying Tet2 haploinsufficiency to mimic the human pre-LHSPC condition and HG stress, in the form of an Ins2Akita/+ mutation, which induces hyperglycemia and type 1 DM, we show that the compound mutant mice developed a lethal form of MPN and/or acute myeloid leukemia (AML). RNA-Seq revealed that this was due in part to upregulation of proinflammatory pathways, thereby generating a feed-forward loop, including expression of the antiapoptotic, long noncoding RNA (lncRNA) Morrbid. Loss of Morrbid in the compound mutants rescued the lethality and mitigated MPN/AML. We describe a mouse model for age-dependent MPN/AML and suggest that hyperglycemia acts as an environmental driver for myeloid neoplasms, which could be prevented by reducing expression levels of the inflammation-related lncRNA Morrbid.


Trends in Malaria in Odisha, India-An Analysis of the 2003-2013 Time-Series Data from the National Vector Borne Disease Control Program.
Although Odisha is the largest contributor to the malaria burden in India, no systematic study has examined its malaria trends. Hence, the spatio-temporal trends in malaria in Odisha were assessed against the backdrop of the various anti-malaria strategies implemented in the state. Using the district-wise malaria incidence and blood examination data (2003-2013) from the National Vector Borne Disease Control Program, blood examination-adjusted time-trends in malaria incidence were estimated and predicted for 2003-2013 and 2014-2016, respectively. An interrupted time series analysis using segmented regression was conducted to compare the disease trends between the pre (2003-2007) and post-intensification (2009-2013) periods. Key-informant interviews of state stakeholders were used to collect the information on the various anti-malaria strategies adopted in the state. The state annual malaria incidence declined from 10.82/1000 to 5.28/1000 during 2003-2013 (adjusted annual decline: -0.54/1000, 95% CI: -0.78 to -0.30). However, the annual blood examination rate remained almost unchanged from 11.25% to 11.77%. The keyinformants revealed that intensification of anti-malaria activities in 2008 led to a more rapid decline in malaria incidence during 2009-2013 as compared to that in 2003-2007 [adjusted decline: -0.83 (-1.30 to -0.37) and -0.27 (-0.41 to -0.13), respectively]. There was a significant difference in the two temporal slopes, i.e., -0.054 (-0.10 to -0.002, p = 0.04) per 1000 population per month, between these two periods, indicating almost a 200% greater decline in the post-intensification period. Although, the seven southern high-burden districts registered the highest decline, they continued to remain in that zone, thereby, making the achievement of malaria elimination (incidence <1/1000) unlikely by 2017. The anti-malaria strategies in Odisha, especially their intensification since 2008, have helped improve its malaria situation in recent years. These successful measures need to be sustained and perhaps intensified further for eliminating malaria from Odisha.


Mapping the dynamics of shear stress-induced structural changes in endothelial cells.
Hemodynamic shear stress regulates endothelial cell biochemical processes that govern cytoskeletal contractility, focal adhesion dynamics, and extracellular matrix (ECM) assembly. Since shear stress causes rapid strain focusing at discrete locations in the cytoskeleton, we hypothesized that shear stress coordinately alters structural dynamics in the cytoskeleton, focal adhesion sites, and ECM on a time scale of minutes. Using multiwavelength four-dimensional fluorescence microscopy, we measured the displacement of rhodamine-fibronectin and green fluorescent protein-labeled actin, vimentin, paxillin, and/or vinculin in aortic endothelial cells before and after onset of steady unidirectional shear stress. In the cytoskeleton, the onset of shear stress increased actin polymerization into lamellipodia, altered the angle of lateral displacement of actin stress fibers and vimentin filaments, and decreased centripetal remodeling of actin stress fibers in subconfluent and confluent cell layers. Shear stress induced the formation of new focal complexes and reduced the centripetal remodeling of focal adhesions in regions of new actin polymerization. The structural dynamics of focal adhesions and the fibronectin matrix varied with cell density. In subconfluent cell layers, shear stress onset decreased the displacement of focal adhesions and fibronectin fibrils. In confluent monolayers, the direction of fibronectin and focal adhesion displacement shifted significantly toward the downstream direction within 1 min after onset of shear stress. These spatially coordinated rapid changes in the structural dynamics of cytoskeleton, focal adhesions, and ECM are consistent with focusing of mechanical stress and/or strain near major sites of shear stress-mediated mechanotransduction.


Oxidative stress in the blood of farm workers following intensive pesticide exposure.
The aim of this study was to evaluate oxidative stress in workers who formulate organophosphate, synthetic pyrethroid and carbamate pesticides. In this survey, blood erythrocytes from a group of 94 pesticide-formulating workers (at least 5-years experience in pest-control in apple and cherry production) and 45 control subjects were examined for oxidative stress parameters. The control group was composed of 45 healthy people living in the same region with no exposure to pesticides. Lipid peroxidation level, catalase, superoxide dismutase and glutathione peroxidase activities in erythrocytes were analysed as biomarkers of oxidative stress. In addition, the acetylcholinesterase activity was measured as a biomarker of toxicity. Results indicated that chronic exposure to organophosphate, synthetic pyrethroid and carbamate pesticides were associated with increased activities of catalase, SOD and lipid peroxidation in erythrocytes (p < 0.05). Acetylcholinesterase activity did not show any significant differences between the two groups (p > 0.05). It is concluded that human chronic exposure to pesticides may result in stimulated antioxidant enzymes.


Transcription Factor STAT3 as a Novel Molecular Target for Cancer Prevention.
Signal Transducers and Activators of Transcription (STATs) are a family of transcription factors that regulate cell proliferation, differentiation, apoptosis, immune and inflammatory responses, and angiogenesis. Cumulative evidence has established that STAT3 has a critical role in the development of multiple cancer types. Because it is constitutively activated during disease progression and metastasis in a variety of cancers, STAT3 has promise as a drug target for cancer therapeutics. Recently, STAT3 was found to have an important role in maintaining cancer stem cells in vitro and in mouse tumor models, suggesting STAT3 is integrally involved in tumor initiation, progression and maintenance. STAT3 has been traditionally considered as nontargetable or undruggable, and the lag in developing effective STAT3 inhibitors contributes to the current lack of FDA-approved STAT3 inhibitors. Recent advances in cancer biology and drug discovery efforts have shed light on targeting STAT3 globally and/or specifically for cancer therapy. In this review, we summarize current literature and discuss the potential importance of STAT3 as a novel target for cancer prevention and of STAT3 inhibitors as effective chemopreventive agents.


Characterization of Inner Retinal Hyperreflective Alterations in Early Cognitive Impairment on Adaptive Optics Scanning Laser Ophthalmoscopy.
To examine inner retinal hyperreflective features on adaptive optics scanning laser ophthalmoscopy (AOSLO) in individuals with early cognitive impairment. In this prospective, cross-sectional study, we enrolled 12 participants with either amnestic mild cognitive impairment (aMCI, n = 10) or early dementia due to Alzheimer's disease (eAD, n = 2) and 12 age-, sex-, and race-matched cognitively normal controls. All participants completed AOSLO imaging of the inner retina. AOSLO montages of the peripapillary area were graded for hyperreflective features including granular membranes, mottled membranes, and nummular features. Regions of interest on AOSLO were compared qualitatively to corresponding optical coherence tomography (OCT) cross sections. OCT was also used to analyze peripapillary retinal nerve fiber layer (RNFL) thickness. Cognitively impaired individuals had a significantly higher number of granular membranes with a larger overall area compared to controls. The proportion of cognitively impaired individuals with two or more granular membranes was 41.7% compared to none in the control group. Granular membrane area was also inversely correlated with cognitive performance on the Montreal Cognitive Assessment. There was no difference between the two groups in terms of other membrane types or RNFL thickness. Individuals with early cognitive impairment related to Alzheimer's show hyperreflective granular membranes on high-resolution imaging, which we hypothesize to be manifestations of inner retinal gliosis. The presence of these subtle hyperreflective membranes may obscure underlying RNFL thinning in these eyes on OCT imaging. The distinctive phenotype of granular membranes surrounding the optic nerve on AOSLO may represent a new potential biomarker of early Alzheimer's.


Rapid in vivo metabolism of a methylether derivative of (+/-)-BW373U86: the metabolic fate of [3H]SNC121 in rats.
Activation of opioid delta receptors produces antinociception without some of the side-effects associated with activation of mu and kappa receptors. (+/-)-BW373U86 [(+/-)-4-[(alpha-R*)-alpha-((2S*,5R*)-4-allyl-2, 5-dimethyl-1-piperazinyl)-3-hydroxybenzyl]-N,N-diethylbenzamide] is a first generation, racemic nonpeptide, partially delta-selective opioid agonist that produces short-lived antinocioception. After systemic, but not central, administration, (+/-)-BW373U86 is also a naltrindole-reversible convulsant. SNC80 [(+)-4-[9-alpha-R)-alpha-((2S,5RO-4-allyl-2, 5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide] is a chiral methylether derivative of (+/-)-BW373U86 with decreased potency, but greater selectivity for the delta-opioid receptor. Like BW373U86, SNC80 produces brief, nonlethal seizures when administered peripherally, albeit at higher doses. Radiolabeling of SNC80 yields a compound with similar pharmacology named [3H]SNC121. [3H]SNC121 was investigated to determine the relationship between its time course of metabolism and the physiological actions of SNC80. The biotransformation of i.p. administered [3H]SNC121 was established in rats in vivo and in vitro via high-performance liquid chromatography analysis of extracted radioactive tissues and fluids. Radioactive equivalents were characterized by their high-performance liquid chromatography retention times and opioid binding activity in rat brain membranes. The kidney, and especially the liver (within 5 min), rapidly metabolize SNC121 to a metabolite with delta-opioid activity coeluting with BW373U86. Direct i.c.v. administration of [3H]SNC121 resulted in minimal metabolism after 1 hr. We conclude that i.p., but not i.c.v., administered [3H]SNC121 can be metabolized rapidly and substantially by the liver to a BW373U86-like compound. The in vivo time course of metabolism after i.p. administration of [3H]SNC121 is consistent with the duration of SNC80 antinociception, and the rapid formation of a BW373U86-like metabolite may also account, in part, for its convulsant properties.


Metabolic remodeling in hypertrophied and failing myocardium: a review.
The energy starvation hypothesis proposes that maladaptive metabolic remodeling antedates, initiates, and maintains adverse contractile dysfunction in heart failure (HF). Better understanding of the cardiac metabolic phenotype and metabolic signaling could help identify the role metabolic remodeling plays within HF and the conditions known to transition toward HF, including "pathological" hypertrophy. In this review, we discuss metabolic phenotype and metabolic signaling in the contexts of pathological hypertrophy and HF. We discuss the significance of alterations in energy supply (substrate utilization, oxidative capacity, and phosphotransfer) and energy sensing using observations from human and animal disease models and models of manipulated energy supply/sensing. We aim to provide ways of thinking about metabolic remodeling that center around metabolic flexibility, capacity (reserve), and efficiency rather than around particular substrate preferences or transcriptomic profiles. We show that maladaptive metabolic remodeling takes multiple forms across multiple energy-handling domains. We suggest that lack of metabolic flexibility and reserve (substrate, oxidative, and phosphotransfer) represents a final common denominator ultimately compromising efficiency and contractile reserve in stressful contexts.


Escalated regeneration in sciatic nerve crush injury by the combined therapy of human amniotic fluid mesenchymal stem cells and fermented soybean extracts, Natto.
Attenuation of inflammatory cell deposits and associated cytokines prevented the apoptosis of transplanted stem cells in a sciatic nerve crush injury model. Suppression of inflammatory cytokines by fermented soybean extracts (Natto) was also beneficial to nerve regeneration. In this study, the effect of Natto on transplanted human amniotic fluid mesenchymal stem cells (AFS) was evaluated. Peripheral nerve injury was induced in SD rats by crushing a sciatic nerve using a vessel clamp. Animals were categorized into four groups: Group I: no treatment; Group II: fed with Natto (16 mg/day for 7 consecutive days); Group III: AFS embedded in fibrin glue; Group IV: Combination of group II and III therapy. Transplanted AFS and Schwann cell apoptosis, inflammatory cell deposits and associated cytokines, motor function, and nerve regeneration were evaluated 7 or 28 days after injury. The deterioration of neurological function was attenuated by AFS, Natto, or the combined therapy. The combined therapy caused the most significantly beneficial effects. Administration of Natto suppressed the inflammatory responses and correlated with decreased AFS and Schwann cell apoptosis. The decreased AFS apoptosis was in line with neurological improvement such as expression of early regeneration marker of neurofilament and late markers of S-100 and decreased vacuole formation. Administration of either AFS, or Natto, or combined therapy augmented the nerve regeneration. In conclusion, administration of Natto may rescue the AFS and Schwann cells from apoptosis by suppressing the macrophage deposits, associated inflammatory cytokines, and fibrin deposits.


Identification of FHL1 as a regulator of skeletal muscle mass: implications for human myopathy.
Regulators of skeletal muscle mass are of interest, given the morbidity and mortality of muscle atrophy and myopathy. Four-and-a-half LIM protein 1 (FHL1) is mutated in several human myopathies, including reducing-body myopathy (RBM). The normal function of FHL1 in muscle and how it causes myopathy remains unknown. We find that FHL1 transgenic expression in mouse skeletal muscle promotes hypertrophy and an oxidative fiber-type switch, leading to increased whole-body strength and fatigue resistance. Additionally, FHL1 overexpression enhances myoblast fusion, resulting in hypertrophic myotubes in C2C12 cells, (a phenotype rescued by calcineurin inhibition). In FHL1-RBM C2C12 cells, there are no hypertrophic myotubes. FHL1 binds with the calcineurin-regulated transcription factor NFATc1 (nuclear factor of activated T cells, cytoplasmic, calcineurin-dependent 1), enhancing NFATc1 transcriptional activity. Mutant RBM-FHL1 forms aggregate bodies in C2C12 cells, sequestering NFATc1 and resulting in reduced NFAT nuclear translocation and transcriptional activity. NFATc1 also colocalizes with mutant FHL1 to reducing bodies in RBM-afflicted skeletal muscle. Therefore, via NFATc1 signaling regulation, FHL1 appears to modulate muscle mass and strength enhancement.


Covalent Linking Greatly Enhances Photoinduced Electron Transfer in Fullerene-Quantum Dot Nanocomposites: Time-Domain Ab Initio Study.
Nonadiabatic molecular dynamics combined with time-domain density functional theory are used to study electron transfer (ET) from a CdSe quantum dot (QD) to the C60 fullerene, occurring in several types of hybrid organic/inorganic nanocomposites. By unveiling the time dependence of the ET process, we show that covalent bonding between the QD and C60 is particularly important to ensure ultrafast transmission of the excited electron from the QD photon-harvester to the C60 electron acceptor. Despite the close proximity of the donor and acceptor species provided by direct van der Waals contact, it leads to a notably weaker QD-C60 interaction than a lengthy molecular bridge. We show that the ET rate in a nonbonded mixture of QDs and C60 can be enhanced by doping. The photoinduced ET is promoted primarily by mid- and low-frequency vibrations. The study establishes the basic design principles for enhancing photoinduced charge separation in nanoscale light harvesting materials.


The Anaplasma ovis genome reveals a high proportion of pseudogenes.
The genus Anaplasma is made up of organisms characterized by small genomes that are undergoing reductive evolution. Anaplasma ovis, one of the seven recognized species in this genus, is an understudied pathogen of sheep and other ruminants. This tick-borne agent is thought to induce only mild clinical disease; however, small deficits may add to larger economic impacts due to the wide geographic distribution of this pathogen. In this report we present the first complete genome sequence for A. ovis and compare the genome features with other closely related species. The 1,214,674 bp A. ovis genome encodes 933 protein coding sequences, the split operon arrangement for ribosomal RNA genes, and more pseudogenes than previously recognized for other Anaplasma species. The metabolic potential is similar to other Anaplasma species. Anaplasma ovis has a small repertoire of surface proteins and transporters. Several novel genes are identified. Analyses of these important features and significant gene families/genes with potential to be vaccine candidates are presented in a comparative context. The availability of this genome will significantly facilitate research for this pathogen.