Ammonium and urea transporter inventory of the selaginella and physcomitrella genomes.
Ammonium and urea are important nitrogen sources for autotrophic organisms. Plant genomes encode several families of specific transporters for these molecules, plus other uptake mechanisms such as aquaporins and ABC transporters. Selaginella and Physcomitrella are representatives of lycophytes and bryophytes, respectively, and the recent completion of their genome sequences provided us with an opportunity for comparative genome studies, with special emphasis on the adaptive processes that accompanied the conquest of dry land and the evolution of a vascular system. Our phylogenetic analysis revealed that the number of genes encoding urea transporters underwent a progressive reduction during evolution, eventually down to a single copy in vascular plants. Conversely, no clear evolutionary pattern was found for ammonium transporters, and their number and distribution in families varies between species. In particular Selaginella, similar to rice, favors the AMT2/MEP family of ammonium transporters over the plant-specific AMT1 type. In comparison, Physcomitrella presents several members belonging to both families.


Influenza virus-specific immunological memory is enhanced by repeated social defeat.
Immunological memory (MEM) development is affected by stress-induced neuroendocrine mediators. Current knowledge about how a behavioral interaction, such as social defeat, alters the development of adaptive immunity, and MEM is incomplete. In this study, the experience of social disruption stress (SDR) prior to a primary influenza viral infection enhanced the frequency and function of the T cell memory pool. Socially stressed mice had a significantly enlarged population of CD8(+) T cells specific for the immunodominant NP366-74 epitope of A/PR/8/34 virus in lung and spleen tissues at 6-12 wk after primary infection (resting memory). Moreover, during resting memory, SDR-MEM mice responded with an enhanced footpad delayed-type hypersensitivity response, and more IFN-gamma-producing CD4(+) T cells were detected after ex vivo stimulation. When mice were rechallenged with A/PR/8/34 virus, SDR-MEM mice terminated viral gene expression significantly earlier than MEM mice and generated a greater D(b)NP(366-74)CD8(+) T cell response in the lung parenchyma and airways. This enhancement was specific to the T cell response. SDR-MEM mice had significantly attenuated anti-influenza IgG titers during resting memory. Similar experiments in which mice were primed with X-31 influenza and challenged with A/PR/8/34 virus elicited similar enhancements in the splenic and lung airway D(b)NP(366-74)CD8(+) T cell populations in SDR-MEM mice. This study demonstrates that the experience of repeated social defeat prior to a primary viral infection significantly enhances virus-specific memory via augmentation of memory T cell populations and suggests that social stressors should be carefully considered in the design and analysis of future studies on antiviral immunity.


Change in the scenery: an ethnographic exploration of crack cocaine use in rural Ohio.
Though still largely perceived as an "inner city" drug, crack cocaine has become increasingly available in many areas of the United States. However, little research has been conducted on the phenomenon of crack cocaine use outside of urban areas. This paper draws on qualitative interviews with 50 recent and former crack-cocaine users to explore patterns of use and distribution in three counties in rural Ohio. The findings show that variable patterns of crack use previously documented among urban crack users are evident in rural areas as well, though these are modified by the context of the rural small town. It is postulated that local social networks are the primary means by which these patterns are both transmitted and translated. Implications of these exploratory findings and possible directions for future research are discussed.


Nonclassical action of retinoic acid on the activation of the cAMP response element-binding protein in normal human bronchial epithelial cells.
Vitamin A (retinol) is essential for normal regulation of cell growth and differentiation. We have shown that the retinol metabolite retinoic acid (RA) induces mucous cell differentiation of normal human tracheobronchial epithelial (NHTBE) cells. However, early biological effects of RA in the differentiation of bronchial epithelia are largely unknown. Here, we showed that RA rapidly activated cAMP response element-binding protein (CREB). However, RA did not use the conventional retinoic acid receptor (RAR)/retinoid X receptor (RXR) to activate CREB. RA activated CREB in NHTBE and H1734 cells in which RARs/RXR were silenced with small interfering RNA (siRNA) targeting RAR/RXR expression or deactivated by antagonist. Inhibition of protein kinase C (PKC) or extracellular regulated kinase (ERK1/2) blocked the RA-mediated activation of CREB. In addition, depletion of p90 ribosomal S6 kinase (RSK) via siRSK1/2 completely abolished the activation, suggesting that PKC, ERK, and RSK are required for the activation. Altogether, this study provides the first evidence that RA rapidly activates CREB transcription factor via PKC, ERK, and RSK in a retinoid receptor-independent manner in normal bronchial epithelial cells. This noncanonical RA signaling pathway may play an important role in mediating early biological effects in the mucociliary differentiation of bronchial epithelia.


[Replication and reassociation kinetics of nuclear matrix DNA from regenerating rat liver].
Fraction 1, containing 65--70% of nuclear DNA (nDNA) was extracted from isolated rat liver nuclei. Then nuclear matrix fraction, containing 30--35% of nDNA was extracted with 1.2 M NaCl. About 1% of nDNA war discovered in residual matrix. Specific activity of residual matrix labelled DNA within 1 and 3 min. after the injection of 14C-orotic acid into the liver portal vein (24 hours after hepatectomy) was respectively in 70 and 50 times higher, and that of matrix DNA--in 45 and 20 times as high as in fraction 1 DNA. Thus, replication begins from nuclear matrix DNA. The latter is enriched with unique sequences by 10%, as compared with total nDNA, and its reassociation kinetics does not change at different stages of the cell cycle (0 and 24 hours after partial hepatectomy). It is suggested, that DNA does not migrate with respect to nuclear matrix under replication.


Characterization of a teleost insulin-like growth factor II (IGF-II) gene: evidence for promoter CCAAT/enhancer-binding protein (C/EBP) sites, and the presence of hepatic C/EBP.
Insulin-like growth factor I (IGF-I) and IGF-II, acting under the regulatory control of growth hormone, are the principal mediators of vertebrate growth. We have previously demonstrated that like humans, but unlike rodents, rainbow trout maintain high hepatic IGF-II messenger RNA levels into adulthood. Here we describe a rainbow trout IGF-II gene with a proximal promoter that contains two CCAAT/enhancer-binding protein (C/EBP) binding sites (TCBS1 and TCBS2). Nuclear proteins corresponding in size to rat C/EBPalpha and C/EBPbeta were detected on Western immunoblots of growth-hormone-treated and mock-treated trout liver extracts. Electrophoretic mobility shift assay of these nuclear extracts further suggests the presence of C/EBPs in trout liver and confirms the ability of TCBS1 to form a complex with trout liver nuclear proteins that is identical in mobility and specificity to that formed by a mammalian consensus CBS construct. In both Western blot and mobility assay results, the growth-hormone-treated trout livers appeared to have a greater accumulation of C/EBP, suggesting a molecular mechanism by which growth hormone can influence the level of serum IGF-II.


Effectiveness of integrated psychological therapy on clinical, neuropsychological, emotional and functional outcome in schizophrenia: a RCT study.
Background: Cognitive impairment is considered a central feature of schizophrenia (SZ) and several rehabilitation treatments have been developed to try to improve cognitive deficits.Aims: The aim of the present study was to analyze the effectiveness of integrated psychological therapy (IPT) compared with a standard treatment (TAU) in two groups of patients with SZ, using a comprehensive testing battery of clinical, cognitive, social cognition and functional outcome domains.Methods: Forty-one patients with SZ were assigned to IPT or TAU groups in a randomized controlled trial (RCT). Psychopathological, neuropsychological, emotional and functional outcome variables were assessed at baseline and after 36 weeks of treatment.Results: The IPT group showed significant improvements than the TAU group regarding clinical and functional outcome variables. Moreover, the IPT group improved significantly in the cognitive domains and emotional functioning. Finally, linear regression has highlighted that the improvement of cognitive variables depends on having done the IPT treatment.Conclusions: IPT seems to be effective in improving clinical, neuropsychological, emotional and functional outcome in chronic SZ inpatients. Further studies would be desirable to deepen the effectiveness of IPT in the field of the psychiatric rehabilitation pointing to the possibility of recovery from mental illness.


Resveratrol-induced changes of the human adipocyte secretion profile.
Enlarged white adipose tissue (WAT) is a feature of obesity and leads to changes in its paracrine and endocrine function. Dysfunction of WAT cells is associated with obesity-associated disorders like type 2 diabetes and cardiovascular diseases. Resveratrol (RSV), a natural polyphenolic compound, mimics beneficial effects of calorie restriction. As such, RSV seems a promising therapeutic target for obesity-associated disorders. The effect of RSV on the human adipokine profile is still elusive. Therefore, a proteomic study together with bioinformatical analysis was performed to investigate the effect of RSV on the secretion profile of mature human SGBS adipocytes. RSV incubation resulted in elevated basal glycerol release and reduced intracellular TG content. This increased intracellular lipolysis was accompanied by profound changes in the adipocyte secretion profile. Extracellular matrix proteins were down-regulated while processing proteins were mostly up-regulated after RSV treatment. Interestingly, RSV induced secretion of proteins protective against cellular stress and proteins involved in the regulation of apoptosis. Furthermore, we found a RSV-induced up-regulation of adiponectin and ApoE accompanied by a down-regulation of PAI-1 and PEDF secretion which may improve anti-inflammatory processes and increased insulin sensitivity. These effects may contribute to alleviate obesity-induced metabolic complications. In addition, two novel RSV-regulated adipocyte-secreted proteins were identified.


Molecularly targeted therapy for malignant glioma.
Malignant gliomas are relatively uncommon but lethal cancers. Despite recent research efforts in cancer therapy, the prognosis of patients with malignant gliomas has remained dismal. Understanding the molecular pathogenesis of glioma may lead to a rational development of new therapies. Despite the genetic heterogeneity of malignant gliomas, common aberrations in the signaling elements of the growth and survival pathways are found. New treatments have emerged to target molecules in these signaling pathways with the goal to increase specific efficacy and minimize toxicity. Monoclonal antibodies and low molecular-weight kinase inhibitors are the most common classes of agents in targeted cancer treatment. Most clinical trials of these agents as monotherapies have failed to demonstrate survival benefit in unselected malignant glioma patient populations. Several mechanisms of treatment failure have been demonstrated. In response, multitargeted kinase inhibitors and combinations of single-targeted kinase inhibitors have been developed to overcome therapeutic resistance. In addition, multimodality combinations of targeted agents with radiation, chemotherapy, or immunotherapy/vaccines may enhance treatment efficacy. Future development of these agents will require advances in discovery and validation of new molecular targets, improvement of therapeutic delivery, and identification of correlative biomarkers. Novel clinical trial designs and endpoints may increase the efficiency of new drug evaluation. In this review, the authors discussed the current understanding of molecular pathogenesis and the development of molecularly targeted therapies in malignant glioma.


The femur-tibia control system of stick insects--a model system for the study of the neural basis of joint control.
In a form of top-down analysis, the femur-tibia control system of stick insects is investigated. Open-loop experiments show that it is mainly velocity-sensitive with an extremely low velocity-threshold, that it possesses a very high gain and that it has only a very small phase reserve and thus works close to instability. The closed-loop system generates catalepsy. The system consists of a single sense organ with approximately 80 sense cells with known characteristics, a small number of interneurones (mainly non-spiking ones) and a small number of motor neurones. The characteristics of the whole system can quantitatively be attributed to the characteristics of its elements. The gain of the loop is state-dependent and the system is 'switched off' during active movements and replaced by a control system with different attributes. It is discussed that most of the characteristics of this system are, at least qualitatively, similar to joint control systems in other animals. Because the described system can be more easily investigated than other systems (especially in vertebrates) it can serve as a model against which more complicated joint control loops may be compared.


Role of Chemical Dynamics Simulations in Mass Spectrometry Studies of Collision-Induced Dissociation and Collisions of Biological Ions with Organic Surfaces.
In this article, a perspective is given of chemical dynamics simulations of collisions of biological ions with surfaces and of collision-induced dissociation (CID) of ions. The simulations provide an atomic-level understanding of the collisions and, overall, are in quite good agreement with experiment. An integral component of ion/surface collisions is energy transfer to the internal degrees of freedom of both the ion and the surface. The simulations reveal how this energy transfer depends on the collision energy, incident angle, biological ion, and surface. With energy transfer to the ion's vibration fragmentation may occur, i.e. surface-induced dissociation (SID), and the simulations discovered a new fragmentation mechanism, called shattering, for which the ion fragments as it collides with the surface. The simulations also provide insight into the atomistic dynamics of soft-landing and reactive-landing of ions on surfaces. The CID simulations compared activation by multiple "soft" collisions, resulting in random excitation, versus high energy single collisions and nonrandom excitation. These two activation methods may result in different fragment ions. Simulations provide fragmentation products in agreement with experiments and, hence, can provide additional information regarding the reaction mechanisms taking place in experiment. Such studies paved the way on using simulations as an independent and predictive tool in increasing fundamental understanding of CID and related processes.


Another way to successfully remove dialysis catheter misplaced into subclavian artery through right internal jugular vein: case report.
For haemodialysis in patients with uraemia, catheterization of the internal jugular or femoral vein is often required to establish access. Puncture with catheterization in the right internal jugular vein (RIJV) is relatively simple, and thus, is the appropriate choice for haemodialysis. However, catheterization at this site can lead to complications, including bleeding at the puncture site. Moreover, in several cases, the haemodialysis catheter (HDC) can be misplaced in the internal carotid artery and subclavian artery, thus making the management troublesome later on. In this article, we report the case of a middle-aged female patient with uraemia wherein a temporal HDC was misplaced into the right subclavian artery during right internal jugular vein catheterization. Instead of conventional surgery and endovascular intervention, the catheter was left in that place for four weeks and subsequently removed directly, followed by local compression for 24 hours. Three days later, a tunnelled cuffed HDC was placed in the RIJV under the guidance of ultrasound and regular haemodialysis was performed.


Influence of 0.2 minimum alveolar concentration of enflurane on the ventilatory response to sustained hypoxia in humans.
To determine the influence of 0.2 minimum alveolar concentration (MAC) of enflurane on the time course of ventilation during sustained hypoxia, we studied 10 healthy adult volunteers with and without enflurane. The following design was used: end-tidal Po2 was maintained at 13.3 kPa for 8 min, at 6.7 kPa for 20 min and at 13.3 kPa for 8 min. End-tidal Pco2 was held constant throughout at 0.67 kPa above the subject's natural value. Control experiments were conducted with no hypoxia imposed. During the experiment subjects breathed via a mouthpiece from an automated gas mixing system which controlled end-tidal values. Enflurane reduced baseline (euoxic) ventilation from 20.9 (SEM 2.0) litre min-1 to 10.1 (1.0) litre min-1 (ANOVA, P < 0.001). Enflurane reduced the acute ventilatory response to hypoxia (AHVR) from 20.1 (3.3) litre min-1 to 5.0 (1.3) litre min-1 (ANOVA, P < 0.01), and the ventilatory off-response at cessation of hypoxia from 11.7 (2.4) litre min-1 to 1.8 (0.5) litre min-1 (ANOVA, P < 0.02). There was no significant difference in hypoxic ventilatory decline (HVD) without and with enflurane (8.9 (2.4) litre min-1 vs 5.5 (1.1) litre min-1; ANOVA, ns). These results confirm that 0.2 MAC of enflurane suppressed the acute ventilatory response to hypoxia, but had no significant effect on the subsequent ventilatory decline during sustained hypoxia.


[Backache in children and adolescents].
Although backpain during childhood is uncommon serious diseases feature high on the list of differential diseases. Often the correct diagnosis in the young is made late or missed in the first instance. During the adolescent period the incidence of backpain rises to include mechanical or osteochondritic conditions and the early problems of a degenerative back such as disc protrusion. It has to be stressed that infection and neoplasma although rare can be a cause of backache at any age. If after thorough investigation no abnormalities has been detected, one should keep in mind that serious mental problems can be the cause of backpain.


Multidrug-Resistant CTX-M-(15, 9, 2)- and KPC-2-Producing Enterobacter hormaechei and Enterobacter asburiae Isolates Possessed a Set of Acquired Heavy Metal Tolerance Genes Including a Chromosomal sil Operon (for Acquired Silver Resistance).
Bacterial resistance to antibiotics is concern in healthcare-associated infections. On the other hand, bacterial tolerance to other antimicrobials, like heavy metals, has been neglected and underestimated in hospital pathogens. Silver has long been used as an antimicrobial agent and it seems to be an important indicator of heavy metal tolerance. To explore this perspective, we searched for the presence of acquired silver resistance genes (sil operon: silE, silS, silR, silC, silF, silB, silA, and silP) and acquired extended-spectrum cephalosporin and carbapenem resistance genes (blaCTX-M and blaKPC) in Enterobacter cloacae Complex (EcC) (n = 27) and Enterobacter aerogenes (n = 8) isolated from inpatients at a general hospital. Moreover, the genetic background of the silA (silver-efflux pump) and the presence of other acquired heavy metal tolerance genes, pcoD (copper-efflux pump), arsB (arsenite-efflux pump), terF (tellurite resistance protein), and merA (mercuric reductase) were also investigated. Outstandingly, 21/27 (78%) EcC isolates harbored silA gene located in the chromosome. Complete sil operon was found in 19/21 silA-positive EcC isolates. Interestingly, 8/20 (40%) E. hormaechei and 5/6 (83%) E. asburiae co-harbored silA/pcoD genes and blaCTX-M-(15,2,or9) and/or blaKPC-2 genes. Frequent occurrences of arsB, terF, and merA genes were detected, especially in silA/pcoD-positive, multidrug-resistant (MDR) and/or CTX-M-producing isolates. Our study showed co-presence of antibiotic and heavy metal tolerance genes in MDR EcC isolates. In our viewpoint, there are few studies regarding to bacterial heavy metal tolerance and we call attention for more investigations and discussion about this issue in different hospital pathogens.


Lysosomal enzyme can bypass the blood-brain barrier and reach the CNS following intranasal administration.
Here we provide the first evidence that therapeutic levels of a lysosomal enzyme can bypass the blood-brain barrier following intranasal administration. α-L-iduronidase (IDUA) activity was detected throughout the brains of IDUA-deficient mice following a single intranasal treatment with concentrated Aldurazyme® (laronidase) and was also detected after intranasal treatment with an adeno-associated virus (AAV) vector expressing human IDUA. These results suggest that intranasal routes of delivery may be efficacious in the treatment of lysosomal storage disorders.


The relationship between trait emotional intelligence and interaction with ostracized others' retaliation.
Regulation of emotions in others is distinct from other activities related to trait emotional intelligence in that only such behavior can directly change other people's psychological states. Although emotional intelligence has generally been associated with prosociality, emotionally intelligent people may manipulate others' behaviors to suit their own interests using high-level capabilities to read and manage the emotions of others. This study investigated how trait emotional intelligence was related to interacting with ostracized others who attempt retaliation. We experimentally manipulated whether two people were simultaneously ostracized or not by using an online ball-tossing game called Cyberball. Eighty university students participated in Cyberball for manipulating ostracism and a "recommendation game," a variation of the ultimatum game for assessing how to interact with others who attempt retaliation, with four participants. After the recommendation game, participants rated their intention to retaliate during the game. People with higher interpersonal emotional intelligence were more likely to recommend that the ostracized other should inhibit retaliation and maximize additional rewards when they have a weaker intention to retaliate. However, they were more likely to recommend that the ostracized other should retaliate against the ostracizers when they have a stronger intention to retaliate. This is the first laboratory study that empirically reveals that people with high interpersonal emotional intelligence influence others' emotions based on their own goals contrary to the general view. Trait emotional intelligence itself is neither positive nor negative, but it can facilitate interpersonal behaviors for achieving goals. Our study offers valuable contributions for the refinement of the trait emotional intelligence concept in the respect of its social function.


The emergency paediatric surgical airway: A systematic review.
Although an emergency surgical airway is recommended in the guidelines for a paediatric cannot intubate, cannot oxygenate (CICO), there is currently no evidence regarding the best technique for this procedure. To review the available literature on the paediatric emergency surgical airway to give recommendations for establishing a best practice for this procedure. Systematic review: Considering the nature of the original studies, a meta-analysis was not possible. MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, Cumulative Index to Nursing and Allied Health Literature, Web of Science, Google Scholar and LILACS databases. Studies addressing the paediatric emergency surgical airway and reporting the following outcomes: time to tracheal access, success rate, complications and perceived ease of use of the technique were included. Data were reported using a Strengths, Weaknesses, Opportunities and Threats analysis. Strengths and Weaknesses describe the intrinsic (dis)advantages of the techniques. The opportunities and threats describe the (dis)advantage of the techniques in the setting of a paediatric CICO scenario. Five studies described four techniques: catheter over needle, wire-guided, cannula or scalpel technique. Mean time for placement of a definitive airway was 44 s for catheter over needle, 67.3 s for the cannula and 108.7 s for the scalpel technique. No time was reported for the wire-guided technique. Success rates were 43 (10/23), 100 (16/16), 56 (87/154) and 88% (51/58), respectively. Complication rates were 34 (3/10), 69 (11/16), 36 (55/151) and 38% (18/48), respectively. Analysis shows: catheter over needle, quick but with a high failure rate; wire-guided, high success rate but high complication rate; cannula, less complications but high failure rate; scalpel, high success rate but longer procedural time. The available data are limited and heterogeneous in terms of reported studies; thus, these results need to be interpreted with caution. The absence of best practice evidence necessitates further studies to provide a clear advice on best practice management for the paediatric emergency surgical airway in the CICO scenario.


In Early Axial Spondyloarthritis, Increasing Disease Activity Is Associated with Worsening of Health-related Quality of Life over Time.
In early axial spondyloarthritis (axSpA), data are lacking about the relationship between disease activity and health-related quality of life (HRQOL). We assessed and quantified the association between change in Ankylosing Spondylitis Disease Activity Score (ASDAS) and HRQOL over time in early axSpA. Baseline and 1-year data of patients with axSpA fulfilling the Assessment of Spondyloarthritis international Society (ASAS) classification criteria from the SPondyloArthritis Caught Early (SPACE) cohort were analyzed. Associations between change in ASDAS and in physical (PCS) or mental component summary (MCS) of the Medical Outcomes Study Short Form-36 were tested by linear regression models. Age, sex, ASAS criteria arm, and blue- versus white-collar work were tested for effect modification. Subsequently, these factors and medication were tested for confounding. There were 161 patients with axSpA [53% male, mean (± SD) age 29.7 (± 7.5) yrs, symptom duration 13.6 (± 7.2) months, HLA-B27-positive 91%, radiographic sacroiliitis 22%] who had ASDAS of 2.5 (± 1.0) and 2.0 (± 0.8), PCS of 28.4 (± 14.3) and 36.9 (± 13.1), and MCS of 48.2 (± 13.8) and 49.3 (± 12.0) at baseline and 1 year, respectively. Per unit increase in ASDAS between baseline and 1 year, PCS worsened by 9.5 points. The same level of disease activity had fewer adverse effects on physical HRQOL in women and white-collar workers. To our knowledge, our data are the first to show that in a broad group of patients with early axSpA, increasing ASDAS is associated with worsening of physical HRQOL, but not mental HRQOL, over time.


[Clinical presentation, course and treatment of recurrent sarcoidosis: followup data].
The paper presents the results of long-term follow-ups of 107 patients with recurrent sarcoidosis registered as having Group VII-IB. In the patients, there was the greatest dissemination of the process in the lung, intrathoracic lymph nodes and other organs and MF MB in the blood, sputum, BAC? Group VIIIB patients need the greatest attention of physicians, treatment involving extracorporeal and physiotherapeutical methods to prevent respiratory failure and disability.


Multiple Nucleocapsid Structural Forms of Shrimp White Spot Syndrome Virus Suggests a Novel Viral Morphogenetic Pathway.
White spot syndrome virus (WSSV) is a very large dsDNA virus. The accepted shape of the WSSV virion has been as ellipsoidal, with a tail-like extension. However, due to the scarcity of reliable references, the pathogenesis and morphogenesis of WSSV are not well understood. Here, we used transmission electron microscopy (TEM) and cryogenic electron microscopy (Cryo-EM) to address some knowledge gaps. We concluded that mature WSSV virions with a stout oval-like shape do not have tail-like extensions. Furthermore, there were two distinct ends in WSSV nucleocapsids: a portal cap and a closed base. A C14 symmetric structure of the WSSV nucleocapsid was also proposed, according to our Cryo-EM map. Immunoelectron microscopy (IEM) revealed that VP664 proteins, the main components of the 14 assembly units, form a ring-like architecture. Moreover, WSSV nucleocapsids were also observed to undergo unique helical dissociation. Based on these new results, we propose a novel morphogenetic pathway of WSSV.


Genetic variation associated with chromosomal aberration frequency: A genome-wide association study.
Chromosomal aberrations (CAs) in human peripheral blood lymphocytes (PBL) measured with the conventional cytogenetic assay have been used for human biomonitoring of genotoxic exposure for decades. CA frequency in peripheral blood is a marker of cancer susceptibility. Previous studies have shown associations between genetic variants in metabolic pathway, DNA repair and major mitotic checkpoint genes and CAs. We conducted a genome-wide association study on 576 individuals from the Czech Republic and Slovakia followed by a replication in two different sample sets of 482 (replication 1) and 1288 (replication 2) samples. To have a broad look at the genetic susceptibility associated with CA frequency, the sample sets composed of individuals either differentially exposed to smoking, occupational/environmental hazards, or they were untreated cancer patients. Phenotypes were divided into chromosome- and chromatid-type aberrations (CSAs and CTAs, respectively) and total chromosomal aberrations (CAtot). The arbitrary cutoff point between individuals with high and low CA frequency was 2% for CAtot and 1% for CSA and CTA. The data were analyzed using age, sex, occupation/cancer and smoking history as covariates. Altogether 11 loci reached the P-value of 10-5 in the GWAS. Replication 1 supported the association of rs1383997 (8q13.3) and rs2824215 (21q21.1) in CAtot and rs983889 (5p15.1) in CTA analysis. These loci were found to be associated with genes involved in mitosis, response to environmental and chemical factors and genes involved in syndromes linked to chromosomal abnormalities. Identification of new genetic variants for the frequency of CAs offers prediction tools for cancer risk in future. Environ. Mol. Mutagen. 60:17-28, 2019. © 2018 Wiley Periodicals, Inc.


Valproic acid inhibits neurosphere formation by adult subventricular cells by a lithium-sensitive mechanism.
The mood stabilizer valproic acid (VPA) decreases neural progenitor proliferation and promotes neurogenesis in the adult hippocampus. However, the effects of VPA on progenitor cells in the adult subventricular zone (SVZ) are not as well characterized. Here we report VPA blocks neurosphere formation and inhibits DNA synthesis in cultured NSCs from the SVZ of adult mice. Inhibition of DNA synthesis is associated with the up-regulation of the differentiation transcription factors Egr1 and Neurod1 and down-regulation of transcription factors associated with "stemness". Co-treatment of VPA with the mood stabilizer lithium antagonizes the anti-proliferative effects of VPA on adult NSCs and abolishes VPA activation of Egr1. Co-treatment of VPA with the MEK1/2 inhibitor PD980589 similarly abolishes Egr1 activation consistent with VPA activation and lithium antagonism of MEK-ERK signaling in adult NSCs. However, Western blot reveals VPA significantly suppresses ERK2 phosphorylation in adult NSCs grown in proliferating culture conditions and that lithium co-treatment does not attenuate this effect. Combined the data indicate VPA inhibition of adult NSC proliferation and activation of Egr1 by VPA, along with the antagonism of these effects by lithium, are the effects of cumulative changes in multiple signaling pathways and are not attributable to a common kinase target.


Membrane currents and cholinergic regulation of K+ current in esophageal smooth muscle cells.
The tight-seal whole cell recording technique with patch pipettes was used to study membrane currents of smooth muscle cells freshly dissociated from the esophagus of cats. Under voltage clamp with K+ in the pipette, depolarizing commands elicited an initial inward current followed by a transient outward current that peaked and then declined to reveal spontaneous outward currents (SOCs). SOCs were evident at -60 mV and more positive potentials. The reversal of SOCs at the K+ equilibrium potential and their suppression by tetraethylammonium chloride lead to the conclusion that they represent the activity of K+ channels. Acetylcholine (ACh) caused reversible contraction of these cells and had two successive effects on membrane currents, causing transient activation of K+ current followed by suppression of SOCs. Both of these effects were blocked by atropine. Consistent with these observations, in current clamp, ACh caused a transient hyperpolarization followed by depolarization. The inward current activated by depolarization was blocked by external Cd2+, consistent with the inward current being a voltage-activated calcium current. Two types of Ca2+ current could be distinguished on the basis of voltage-activation range, time course of inactivation and "run-down" during whole cell recording.


Identification of Missense Extracellular Matrix Gene Variants in a Large Glaucoma Pedigree and Investigation of the N700S Thrombospondin-1 Variant in Normal and Glaucomatous Trabecular Meshwork Cells.
Primary open-angle glaucoma (POAG) is a complex heterogeneous disease. While several POAG genes have been identified, a high proportion of estimated heritability remains unexplained. Elevated intraocular pressure (IOP) is a leading POAG risk factor and dysfunctional extracellular matrix (ECM) in the trabecular meshwork (TM) contributes to elevated IOP. In this study, we sought to identify missense variants in ECM genes that correlate with ocular hypertensive POAG. Whole-genome sequencing was used to identify genetic variants in five members of a large POAG family (n = 68) with elevated IOP. The remaining family members were screened by Sanger sequencing. Unrelated normal (NTM) and glaucomatous (GTM) cells were sequenced for the identified variants. The ECM protein levels were determined by Western immunoblotting and confocal and electron microscopy investigated ECM ultrastructural organization. Three ECM gene variants were significantly associated with POAG or elevated IOP in a large POAG pedigree. These included rs2228262 (N700S; thrombospondin-1 (THBS1, TSP1)), rs112913396 (D563 G; collagen type VI, alpha 3 (COL6A3)) and rs34759087 (E987K; laminin subunit beta 2 (LAMB2)). Screening of unrelated TM cells (n = 27) showed higher prevalence of the THBS1 variant but not the LAMB2 variant, in GTM cells (39%) than NTM cells (11%). The rare COL6A3 variant was not detected. TSP1 protein was upregulated and COL6A3 was down-regulated in TM cells with N700S subject to mechanical stretch, an in vitro method that mimics elevated IOP. Immunofluorescence showed increased TSP1 immunostaining in cell strains with N700S compared to wild-type TM cells. Ultrastructural studies showed ECM disorganization and altered collagen type VI distribution in GTM versus NTM cells. Our results suggest that missense variants in ECM genes may not cause catastrophic changes to the TM, but over many years, subtle changes in ECM may accumulate and cause structural disorganization of the outflow resistance leading to elevated IOP in POAG patients.


HIF-1α repairs degenerative chondrocyte glycolytic metabolism by the transcriptional regulation of Runx2.
HIF-1α and Runx2 expression usually increase in chondrocytes (CHs) during osteoarthritis (OA), which involves the changes in glycolytic metabolism. However, the molecular regulation of HIF-1α related to the CHs glycolytic metabolism is still unclear. In this study, we aimed to reveal the mediation of HIF-1α by Runx2 and its effect on the glycolytic metabolism of degenerative CHs. The expression of HIF-1α, Runx2, and the degenerative markers of CHs in both natural conditions from the OA patients and IL-1β treated in vitro model was analyzed by a Western blot or real-time polymerase chain reaction (RT-PCR). The glycolytic metabolism was determined by the intracellular glucose uptake and adenosine triphosphate (ATP) generation. Transfection of siRNA coding HIF-1α or Runx2 was used to clear the function between HIF-1α and Runx2 in the glycolytic metabolism of degenerated CHs caused by IL-1β. Chromatin immunoprecipitation (ChIP) and Luciferase reporter gene assay were used to verify the Runx2 protein binds to the promoter of HIF-1α and promote its expression. HIF-1α and Runx2 were increased, and glucose uptake and ATP generation were decreased in the degenerative CHs from both OA and IL-1β conditions. Under the stimulation of IL-1β, Runx2 silencing rejected the upregulation of HIF-1α and further aggravated the glycolytic metabolism. When HIF-1α was silenced, the glycolytic metabolism of CHs was also suppressed. Besides, Runx2 protein could regulate HIF-1α expression in the transcriptional level by binding to its promoter. OHIF-1α plays a role in the self-repair of the glycolytic metabolism of degenerative CHs via the transcriptional regulation of Runx2.


Synthesis and Testing of Analogs of the Tuberculosis Drug Candidate SQ109 against Bacteria and Protozoa: Identification of Lead Compounds against Mycobacterium abscessus and Malaria Parasites.
SQ109 is a tuberculosis drug candidate that has high potency against Mycobacterium tuberculosis and is thought to function at least in part by blocking cell wall biosynthesis by inhibiting the MmpL3 transporter. It also has activity against bacteria and protozoan parasites that lack MmpL3, where it can act as an uncoupler, targeting lipid membranes and Ca2+ homeostasis. Here, we synthesized 18 analogs of SQ109 and tested them against M. smegmatis, M. tuberculosis, M. abscessus, Bacillus subtilis, and Escherichia coli, as well as against the protozoan parasites Trypanosoma brucei, T. cruzi, Leishmania donovani, L. mexicana, and Plasmodium falciparum. Activity against the mycobacteria was generally less than with SQ109 and was reduced by increasing the size of the alkyl adduct, but two analogs were ∼4-8-fold more active than SQ109 against M. abscessus, including a highly drug-resistant strain harboring an A309P mutation in MmpL3. There was also better activity than found with SQ109 with other bacteria and protozoa. Of particular interest, we found that the adamantyl C-2 ethyl, butyl, phenyl, and benzyl analogs had 4-10× increased activity against P. falciparum asexual blood stages, together with low toxicity to a human HepG2 cell line, making them of interest as new antimalarial drug leads. We also used surface plasmon resonance to investigate the binding of inhibitors to MmpL3 and differential scanning calorimetry to investigate binding to lipid membranes. There was no correlation between MmpL3 binding and M. tuberculosis or M. smegmatis cell activity, suggesting that MmpL3 is not a major target in mycobacteria. However, some of the more active species decreased lipid phase transition temperatures, indicating increased accumulation in membranes, which is expected to lead to enhanced uncoupler activity.


Increased beta-adrenoceptor binding capacity is associated with blunted beta-adrenoceptor-mediated cardiovascular responses in essential hypertension.
Age and elevated blood pressure are associated with blunted beta-adrenoceptor-mediated cardiovascular effects. To investigate a possible relationship between beta-adrenoceptor cardiovascular function and receptor density, beta-adrenoceptor binding capacity in mononuclear leucocytes and cardiac isoproterenol sensitivity were compared in 12 essential hypertensive and 17 normotensive subjects of comparable age. The bolus dose of isoproterenol which increased heart rate by 25 beats/min (CD25) as well as plasma norepinephrine, epinephrine, and renin activity were measured. In a radioreceptor assay using [3H]dihydroalprenolol, the antagonist binding capacity (Bmax) and the affinity constant (KD) of mononuclear leucocytes were determined. Bmax in patients was higher than in normotensive subjects (66.8 +/- 4.1 versus 48.0 +/- 3.8 SEM fmol/mg, p less than 0.05), and KD was identical in both groups. In hypertensive patients, Bmax correlated positively with age (r = 0.639, p less than 0.05) and CD25 (r = 0.593, p less than 0.05) and negatively with plasma renin activity (r = 0.679, p less than 0.05), while in normotensive subjects, Bmax correlated with CD25 only (r = 0.615, p less than 0.05). Thus, in patients with essential hypertension, a decrease in cardiac isoproterenol sensitivity and plasma renin activity is associated with an age-related increase in antagonist binding capacity. This suggests a defect in the membrane coupling of the beta-adrenoceptor effector system distal to the receptor recognition site in patients with essential hypertension.


ATP hydrolysis is required for relocating cohesin from sites occupied by its Scc2/4 loading complex.
The Cohesin complex that holds sister chromatins together until anaphase is comprised of three core subunits: Smc1 and Smc3, two long-rod-shaped proteins with an ABC-like ATPase head (nucleotide-binding domain [NBD]) and a dimerization domain linked by a 50 nm long intramolecular antiparallel coiled-coil, and Scc1, an α-kleisin subunit interconnecting the NBD domains of Smc1 and Smc3. Cohesin's stable association with chromosomes is thought to involve entrapment of chromatin fibers by its tripartite Smc1-Smc3-Scc1 ring via a poorly understood mechanism dependent on a separate Scc2/4 loading complex. A key issue concerns where entrapment initially takes place: at sites where cohesin is found stably associated or at distinct "loading" sites from which it translocates. In this study, we find transition state mutant versions (Smc1E1158Q and SmcE1155Q) defective in disengagement of their nucleotide binding domains (NBDs), unlike functional cohesin, colocalize with Scc2/4 at core centromeres, sites that catalyze wild-type cohesin's recruitment to sequences 20 kb or more away. In addition to Scc2/4, the unstable association of transition state complexes with core centromeres requires Scc1's association with Smc1 and Smc3 NBDs, ATP-driven NBD engagement, cohesin's Scc3 subunit, and its hinge domain. We propose that cohesin's association with chromosomes is driven by two key events. NBD engagement driven by ATP binding produces an unstable association with specific loading sites like core centromeres, whereas subsequent ATP hydrolysis triggers DNA entrapment, which permits translocation along chromatin fibers.


Effects of pregnancy, parturition, and anal sphincter transection on function of the external anal sphincter in an animal model.
To estimate the effects of pregnancy, parturition, and anal sphincter laceration (with repair) on external anal sphincter morphology and neurophysiology and to define the time course of these effects after injury. Within 4 hours of vaginal delivery, 80 rats underwent either sham or anal sphincter laceration with repair. After 3 days, 3 weeks, and 3 and 6 months (n=20 for each time point), animals were killed, and the anal sphincter complexes dissected and removed for neurophysiologic studies. Twitch tension, peak tetanic force, fatigue, and maximal electrical field-stimulated force generation were determined. Sphincters were then fixed and serially sectioned (5-micrometer thickness) at 100-micrometer intervals for histologic analysis. Maximal electrical field-stimulated force generation, maximal tetanic contraction, and twitch tension were decreased in the external anal sphincter 3 days after anal sphincter laceration with repair compared with sham-operated parturient rats (3.3 g compared with 11.6 g, 4.5 g compared with 14.5 g, and 0.6 g compared with 2.0 g, respectively, all P<.02). Increased fatigability of the sphincter muscle was observed in all newly parturient rats-sham and anal sphincter laceration with repair; recovery occurred in the shams by 3 months. A gradual recovery occurred in all these neurophysiologic measures, with no significant differences between anal sphincter laceration with repair and shams by 6 months postpartum. Repaired anal sphincter transection in periparturient animals results in short-term severe compromise of neurophysiologic function of the external anal sphincter. Over time, however, force generation recovers and approximates that of postpartum rats with intact anal sphincters.


The prophylaxis and treatment potential of supplements for COVID-19.
The global impact of the new coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), infection that caused COVID-19 has been evident in the last few months from the unprecedented socioeconomic disruption to more than 600,000 deaths. The lack of vaccine and effective therapeutic agents for the disease prompted world-wide effort to test those antiviral therapeutics already in use for other diseases. Another interesting approach has been based on the pathological sequel of the disease that involve severe inflammatory reaction (or the cytokine storm) associated with pneumonia in critically ill patients. This article outlines the prophylaxis therapeutic potential of supplements vitamins and micronutrients in COVID-19. By ameliorating the inflammatory and oxidative stress associated with the disease and some direct antiviral effects, the application of these agents as adjuvants and other alternative approaches are discussed. Available clinical trials including those currently registered on these supplements are scrutinized.


Arbitrarily primed PCR, ribotyping, and plasmid pattern analysis applied to investigation of a nosocomial outbreak due to Enterobacter cloacae in a neonatal intensive care unit.
In December 1992, Enterobacter cloacae was isolated from the oropharynx and respiratory tract of six ventilated neonates hospitalized in the intensive care unit (ICU) of our hospital. To establish the spread of the outbreak, 41 strains of E. cloacae were analyzed for genotypic markers by three methods: plasmid profile analysis, ribotyping with EcoRI or PvuII endonuclease, and arbitrarily primed (AP) PCR. The tested strains included 12 isolates from the 6 epidemic cases, 4 isolates from the respiratory tract of 4 children hospitalized in other wards during the same period, 13 isolates from 12 children hospitalized in pediatric units before or after the outbreak, and 12 epidemiologically unrelated isolates. Ribotyping and AP PCR demonstrated that each of the last 12 strains exhibited distinct genomic patterns, as did each of the strains isolated from neonates hospitalized before or after the epidemic peak. Conversely, two clones of strains were found among the isolates recovered in December, with concordant results being obtained by the three typing methods: the first clone included seven strains from five ventilated children in the ICU and two children from another ward; another clone was shared by one neonate in the ICU and an infant from another ward. These results indicate that ribotyping and AP PCR-the latter applied, to our knowledge, for the first time to the genotypic analysis of E. cloacae--represent very discriminatory tools for the investigation of nosocomial outbreaks caused by this species.


The deletion of 22q13 region in both intracranial and spinal meningiomas in a patient (case report).
We present a 69 year old man with two simultaneous meningiomas in different compartment of neural axis, in both of which 22q13 locus is lost. Histologically the two tumours appeared to be different; meningotheliomatous and transitional with psammoma bodies, respectively. No numerical or structural chromosome abnormalities were seen in karyotype analysis of the cultured spinal and cranial meningioma samples. Since long arm structural aberrations and/or whole loss of chromosome 22 are frequently reported abnormalities of meningiomas, the tumours were also analysed by fluorescence in situ hybridisation (FISH) with different colour-labelled probes in respect to relevant chromosome. The metaphases and interphase nuclei of the samples were evaluated by the combined biotinylated 22q11 and digoxigenin-labelled 22q13 locus specific FISH probes, and 22q13 deletion was revealed in both of spinal and cranial tumour cells. In conclusion, since both tumours from the presented case show the same genetic alterations, multiplicity may be derived from the same clone of cells, and support the theory of development of multiple meningiomas from the spreading of tumour cells via cerebrospinal fluid as a possible mechanism.


Sulfur diagenesis in everglades peat and origin of pyrite in coal.
The pattern of sulfur transformation in peat across the Everglades basin indicates that pyrite formation in organic-rich swamps depends on the use of organic oxysulfur compounds in dissimilatory respiration by sulfur-reducing bacteria. This paragenesis explains the primary distribution of sulfur compounds in low-sulfur coals and possibly in most coals and many organic-rich soils and sediments. It also accounts for the occurrence of framboidal pyrite bound in fossil tissue in coal and sediments.


Gender Equality in Neurosurgery and Strategic Goals Toward a More Balanced Workforce.
The Women in Neurosurgery (WINS) and the American Association of Neurological Surgeons published a white paper in 2008 setting an ambitious goal for women to comprise 20% of neurosurgery residents by 2012 and 20% of practicing neurosurgeons by 2020. Although there has been steady progress, we have fallen short of these benchmarks. We take this opportunity to look back at the accomplishments made over the past decade and provide an update on our present status. We evaluate current barriers toward progress and propose new goals, highlighting the systemic changes necessary to accomplish them. We propose the following updated recommendations to recruit and retain diverse talent into the neurosurgical workforce. (1) Neurosurgical departments and societies should provide diverse, early formal mentorship opportunities for medical students, residents, and junior faculty members. (2) Parental leave policies must be delineated, promoted, and enforced for all neurosurgeons, with greater awareness of internal discrimination and normalization of the discussion surrounding this topic. (3) We need to strive for compensation equity, with transparency in compensation mechanisms and regular assessment of compensation metrics. (4) Departments and institutions must have a zero-tolerance policy for sexual harassment and discrimination and establish a safe reporting structure. Finally, we propose attainable benchmarks toward achieving gender balance in the neurosurgical workforce, with a goal for women to comprise 30% of the entering residency class by 2030 and to comprise 30% of practicing neurosurgeons by 2038. We hope that this will guide further progress toward our future of building a balanced workforce.


Increased tau burden in the cortices of progressive supranuclear palsy presenting with corticobasal syndrome.
The objective of this study is to better define the pathological characteristics of pathologically proven progressive supranuclear palsy (PSP) presenting with the corticobasal syndrome (CBS). PSP is characterized by early falls, vertical supranuclear ophthalmoplegia, and axial rigidity, whereas asymmetric limb features, including rigidity, bradykinesia, apraxia, alien limb phenomena, and cortical sensory loss are characteristic of CBS. We investigated clinicopathological characteristics of 5 cases of PSP that presented with CBS (CBS-PSP). Comprehensive pathological analysis was undertaken to determine the presence of concomitant pathological processes as well as quantitative tau burden in cortical regions of CBS-PSP, compared with 8 typical PSP cases (Typ-PSP). The clinical features in the CBS-PSP cases included asymmetrical features, apraxia, alien limb phenomena, and progressive aphasia. All cases had Parkinsonism, and vertical supranuclear ophthalmoplegia was noted in all but 1 case of CBS-PSP. Secondary neuropathological diagnoses included argyrophilic grain disease (AGD) in 1 of the 8 cases of Typ-PSP, whereas Alzheimer's disease (AD), Lewy body disease, AGD, and vascular disease was found in 3 cases of CBS-PSP. Image analysis of cortical tau burden performed in 8 Typ-PSP and 3 CBS-PSP cases revealed a significant increased tau burden in mid-frontal and inferior-parietal cortices in the CBS-PSP cases. This study demonstrates that when PSP presents as CBS, it is most likely due to either a concurrent cortical pathology from a secondary process such as AD or from the primary pathology of PSP extending into cortical areas that are primarily and commonly affected in CBD.


[Effects of lung- and liver-derived biomatrices on the attachment and growth of tumor cells].
The attachment and growth property of a human carcinoma cell line (Anip), which has high pulmonary metastatic potential, were investigated in vitro on the lung- or liver-derived biomatrix in culture medium supplemented with serum (SSM) or free of serum (SFM). Either in SFM or SSM, the carcinoma cells showed a higher rate of adhesion to the lung biomatrix than that to the liver biomatrix, In SFM, cells plated on plastic plates ceased proliferating and died; whereas those on the biomatrices, particularly the cells on the lung biomatrix survived and proliferated. Moreover, in SSM, carcinoma cells on the lung biomatrix had the highest clonal growth efficiency. These results indicate that biomatrices may provide conditions favourable for cell growth, and may compensate in part the deficiency of nutrition and growth factors when serum is absent in the culture medium. The different influences exerted on the growth and attachment rate of tumor cells by biomatrices derived from different organs also help to clarify the mechanism of organ specificity of tumor metastasis.


Development of a GC-MS method for the simultaneous analysis of latent fingerprint components.
Latent fingerprint residue is a complex mixture of organic and inorganic compounds. A full understanding of the composition of this mixture and how it changes after deposition is lacking. Three solvent systems were compared for the simultaneous extraction and derivatization with ethyl chloroformate of selected amino and fatty acids from a nonporous substrate (Mylar for subsequent analysis by gas chromatography-mass spectrometry. A solvent system comprised of sodium hydroxide, ethanol, and pyridine was found to be the most effective. This method was applied to the analysis of latent fingerprint residue deposited on Mylar and preliminary data are presented. Twelve amino acids (e.g., serine, glycine, and aspartic acid) and 10 fatty acids (e.g., tetradecanoic, hexadecanoic, and octadecanoic acids) were identified. The potential application of this method to further the understanding of latent fingerprint chemistry has been demonstrated.


Evaluating Family Group Conferencing: Towards a meaningful research methodology.
There is discussion on the most appropriate research methodology to examine the efficacy of Family Group Conferencing (FGC). Randomised controlled trials (RCTs), despite their pitfalls, are considered by many to be the 'golden standard', but the argument is not compelling. In this paper, the theory on programme evaluation is discussed which offers an alternative methodology to study FGC. It is argued that reaching a comprehensive image of truth in the social sciences is never within reach. A RCT is an abstraction of reality, it only provides a partial image of the complex reality of families and the impact that FGC has on safety issues and the quality of their lives. Moreover, the rigour of a study depends heavily on the researcher's interpretative skills. In studying the efficacy of a complex intervention, such as FGC, it is a challenge to provide a valid and reliable picture of its impact. The context of such a conference, where the lifeworld of families constantly interacts with the system world of professionals, is characterised by multiplicity, polyvalence and interference. The methodology used to examine the efficacy of FGC should meet this 'interplexitiy'.


Characterizing and Comparing Per- and Polyfluoroalkyl Substances in Commercially Available Biosolid and Organic Non-Biosolid-Based Products.
There is increasing concern over the presence of per- and polyfluoroalkyl substances (PFAS) in biosolids, while sales in commercially available biosolid-based products used as soil amendments are also increasing. Here, the occurrence of 17 perfluoroalkyl acids (PFAAs) present in 13 commercially available biosolid-based products, six organic composts (manure, mushroom, peat, and untreated wood), and one food and yard waste compost were studied. The PFAA concentration ranges observed are as follows: biosolid-based products (9.0-199 μg/kg) > food and yard waste (18.5 μg/kg) > other organic products (0.1-1.1 μg/kg). Analysis of 2014, 2016, and 2018 bags produced from one product line showed a temporal decrease in the total PFAAs (181, 101, and 74 μg/kg, respectively). The total oxidizable precursor (TOP) assay revealed the presence of PFAA precursors in the biosolid-based products at much higher levels, when the soluble carbon was removed by the ENVI-Carb clean-up prior to the TOP assay. Time-of-flight mass spectrometry confirmed the presence of three sulfonamides, two fluorotelomer sulfonates, and several polyfluoroalkyl phosphate diesters. Pore-water concentrations of water-saturated products were primarily of short-chain PFAAs and increased with increasing PFAA concentrations in the products. A strong positive log-linear correlation between organic carbon (OC)-normalized PFAA partition coefficients and the number of CFn units indicates that OC is a good predictor of PFAA release concentrations.


Medullary thyroid carcinoma: targeted therapies and future directions.
Medullary thyroid cancer (MTC) is a rare neuroendocrine neoplasm that accounts for approximately 5% of all thyroid malignancies. The natural history of MTC is characterized by early lymph node and distant metastases, making complete surgical cure often impossible. Conventional chemotherapy and external beam radiation have been largely ineffective in altering the natural history of MTC. Therefore, there is a great need to develop novel therapeutic strategies to affect symptom control and reduce tumor burden in patients with widely disseminated disease. Here, we review several pathways which have been shown to be vital in MTC tumorigenesis and focus on the pathways of interest for which targeted drug therapies are currently being developed.


Regional differences in dynamic cerebral autoregulation in the healthy brain assessed by magnetic resonance imaging.
A novel method is described for mapping dynamic cerebral blood flow autoregulation to assess autoregulatory efficiency throughout the brain, using magnetic resonance imaging (MRI). Global abnormalities in autoregulation occur in clinical conditions, including stroke and head injury, and are of prognostic significance. However, there is limited information about regional variations. A gradient-echo echo-planar pulse sequence was used to scan the brains of healthy subjects at a rate of 1 scan/second during a transient decrease in arterial blood pressure provoked by a sudden release of pressure in bilateral inflated thigh cuffs. The signal decrease and subsequent recovery were analyzed to provide an index of autoregulatory efficiency (MRARI). MRI time-series were successfully acquired and analyzed in eleven subjects. Autoregulatory efficiency was not uniform throughout the brain: white matter exhibited faster recovery than gray (MRARI = 0.702 vs. 0.672, p = 0.009) and the cerebral cortex exhibited faster recovery than the cerebellum (MRARI = 0.669 vs. 0.645, p = 0.016). However, there was no evidence for differences between different cortical regions. Differences in autoregulatory efficiency between white matter, gray matter and the cerebellum may be a result of differences in vessel density and vasodilation. The techniques described may have practical importance in detecting regional changes in autoregulation consequent to disease.


Team-based learning enhances long-term retention and critical thinking in an undergraduate microbial physiology course.
We used team-based learning to improve comprehension and critical thinking of students in an undergraduate microbial metabolism-physiology course. The course used well-known bacterial pathways to highlight themes of energy conservation and biodegradation. Prior to the introduction of team-based learning, student recall of this information was poor and students had difficulty extrapolating information to new organisms. Initially, individual and group quizzes were added to promote problem-solving and critical-thinking skills. This significantly improved student attitudes about the amount of information they learned and whether the instructor promoted critical thinking. However, retention of the material as judged by final examination scores was still poor. In the next year, two challenging projects were added to the course to complement the above themes: (i) postulating a pathway for the metabolism of a substrate by a bacterium, and (ii) modifying the current model for anaerobic sulfate reduction by incorporating recent genetic information. The inclusion of the team projects significantly improved final examination scores compared to the previous year without team projects. Overall, team-based learning with challenging projects improved the students' comprehension and retention of information, critical thinking, and attitudes about the course and focused student-instructor interactions on learning rather than grades.


Endothelium-denuded pulmonary resistance arteries from the fetal lamb: preparation and response to vasoactive agents.
A method was developed for preparing endothelium-denuded resistance arteries (mean internal diameter, 176 microns) from the term fetal lung to be used for tests in vitro. The endothelium was removed mechanically by passing a sandpaper-polished cat whisker through the lumen. Ultrastructurally, the preparation had a well-preserved internal elastic lamina which was facing the lumen without any endothelium superimposed. Its response to contractile agents (excess potassium, a thromboxane A2 analogue, endothelin-1) tended to be greater compared to the intact artery. Conversely, the relaxation to acetylcholine was abolished in the endothelium-denuded artery and, in its place, a modest contraction ensued. Sodium nitroprusside relaxation, unlike acetylcholine relaxation, was affected insignificantly by endothelium denudation. This preparation, in combination with the preparation of the intact resistance artery, is a useful, new tool for studying local factors responsible for pulmonary hemodynamic control in utero and through the transitional period at birth.


Pediatric vs. Adult Prodrome and Postdrome: A Window on Migraine Pathophysiology?
Few studies have been conducted on the prodromal and postdromal phases of the migraine attack in children and adolescents. Using a questionnaire, we found that 67% of 103 children and adolescents with migraine reported at least one prodromal symptom, with a mean number per subject of 1.8 (median 2.2). The most frequently reported prodromal symptoms were face changes, fatigue and irritability. In pediatric patients selected as having prodrome, fatigue, mood change and neck stiffness were the most frequently reported prodromal symptoms. Using a different design, Laurell et al. found that 71% of 137 pediatric patients reported at least one prodromal symptom with a mean number per subject of 1.9 ± 2.0. Studying postdrome was fraught with unexpected difficulties as our preliminary research showed. Patients reported 2 groups of symptoms occurring during the resolution phase of the headache: symptoms whose onset was before headache cessation and were persisting after it, and symptoms whose onset was after headache cessation. We referred to the former as persistent symptoms and to the latter as true postdromes. Ninety-one per cent of patients reported persistent symptoms, with a mean of 6.0 and a median of 2, asthenia, pallor, cognitive difficulties, anorexia, somnolence, and nausea being the more frequently reported. True postdromes were reported by 82% of patients, with a mean of 2.6 and a median of 2, thirst, somnolence, visual disturbances, food craving, paraesthesias, and ocular pain being the most frequent reported. Interestingly, several prodromal and postdromal symptoms are also encountered during the aura classic and/or accompany the headache phase. Functional imaging in migraine has showed that the activations in areas such as hypothalamus or brainstem may begin before headache onset and/or persist after headache relief. Thus, one may wonder whether prodromal and postdromal symptoms may indicate the involvement of the limbic system, dopaminergic pathways, the hypothalamus and the brainstem. Differences between children, adolescents and adults might contribute to the understanding of migraine neurobiology.


Suppression of experimental glomerulonephritis by the interleukin-1 receptor antagonist: inhibition of intercellular adhesion molecule-1 expression.
Interleukin-1 is a proinflammatory cytokine produced in glomerulonephritis. Blocking the action of interleukin-1 by the administration of the interleukin-1 receptor antagonist (IL-1ra) has been shown to prevent renal function impairment, reduce glomerular injury, inhibit leukocyte infiltration, and suppress tubulointerstitial damage in experimental antiglomerular basement membrane disease. A key mechanism in the entry of leukocytes into the kidney is the interaction between the interleukin-1 inducible intercellular adhesion molecule-1 (ICAM-1; CD54) and lymphocyte function-associated antigen-1 (CD11a/CD18). Therefore, this study investigated whether the inhibition of this mechanism was the means by which IL-1ra suppressed leukocyte infiltration in rat accelerated antiglomerular basement membrane glomerulonephritis. Disease was induced in two groups of six rats; animals were treated by constant sc infusion of recombinant human IL-1ra or saline from the initiation of disease until being euthanized 14 days later. In saline-treated animals, there was marked up-regulation of ICAM-1 in the glomerulus and interstitium, In which was associated with leukocyte infiltration. In particular, focal accumulation of CD11a+ and CD18+ cells was apparent in areas of tubulointerstitial damage exhibiting intense ICAM-1 expression. IL-1ra treatment partially reduced glomerular ICAM-1 expression and leukocyte infiltration. However, IL-1ra treatment resulted in a dramatic inhibition of interstitial ICAM-1 expression, interstitial leukocyte infiltration, and tubulointerstitial damage. In conclusion, this study has shown that interleukin-1 is a major inducer of ICAM-1 expression within the renal tubulo-interstitium--a process associated with focal leukocyte infiltration and tubulointerstitial damage.(ABSTRACT TRUNCATED AT 250 WORDS)


Oxidoreductive cellulose depolymerization by the enzymes cellobiose dehydrogenase and glycoside hydrolase 61.
Several members of the glycoside hydrolase 61 (GH61) family of proteins have recently been shown to dramatically increase the breakdown of lignocellulosic biomass by microbial hydrolytic cellulases. However, purified GH61 proteins have neither demonstrable direct hydrolase activity on various polysaccharide or lignacious components of biomass nor an apparent hydrolase active site. Cellobiose dehydrogenase (CDH) is a secreted flavocytochrome produced by many cellulose-degrading fungi with no well-understood biological function. Here we demonstrate that the binary combination of Thermoascus aurantiacus GH61A (TaGH61A) and Humicola insolens CDH (HiCDH) cleaves cellulose into soluble, oxidized oligosaccharides. TaGH61A-HiCDH activity on cellulose is shown to be nonredundant with the activities of canonical endocellulase and exocellulase enzymes in microcrystalline cellulose cleavage, and while the combination of TaGH61A and HiCDH cleaves highly crystalline bacterial cellulose, it does not cleave soluble cellodextrins. GH61 and CDH proteins are coexpressed and secreted by the thermophilic ascomycete Thielavia terrestris in response to environmental cellulose, and the combined activities of T. terrestris GH61 and T. terrestris CDH are shown to synergize with T. terrestris cellulose hydrolases in the breakdown of cellulose. The action of GH61 and CDH on cellulose may constitute an important, but overlooked, biological oxidoreductive system that functions in microbial lignocellulose degradation and has applications in industrial biomass utilization.


Does gene expression in laryngeal subsites differ between patients with laryngopharyngeal reflux and controls?
To identify laryngeal mRNA gene changes in patients with laryngopharyngeal reflux (LPR). Laryngeal biopsies from non-smoking LPR patients (n=10; Reflux Symptom Index (RSI) >12 and a Reflux Finding Score (RFS) >6) and controls (n=9; RSI <12 and RFS <6) were collected from four subsites (true vocal cord, false vocal cord, medial arytenoid and posterior commissure) of the larynx. qRT-PCR analyses were conducted on 20 reflux- and inflammation-related genes, including interleukins 6 and 8, cytokeratins 8 and 14, mucin genes MUC1, MUC2, MUC3B, MUC4, MUC5B, MUC6 and MUC7 and carbonic anhydrase III. Statistical analysis (Mann-Whitney U test) compared gene expression levels between LPR and control groups at each subsite. Site-specific differences in squamous metaplasia and gene expression were noted in LPR patients, with the majority present in the medial arytenoid region. Significant.differences were noted in genes related to mucosal defence and inflammation, including CRNN, CD1d, TGFβ-1, MUC2, MUC5B and CDH1. Whilst the posterior commissure is commonly identified as the area demonstrating the most significant macroscopic change in LPR, the histological changes and genes assessed here showed more pronounced LPR associated differences in the medial arytenoid. We identified differences in expression of mucin genes, cytokeratin-14 and molecular markers of inflammation. Whilst some of these changes may be metaplasia-related, further evaluation of the mRNA expression of these genes may provide a useful biomarker panel for diagnosis and therapeutic monitoring of LPR.


A CRM1-mediated nuclear export signal is essential for cytoplasmic localization of neurogenin 3 in neurons.
Neurogenin 3 (Ngn3), a proneural gene, regulates dendritogenesis and synaptogenesis in mouse hippocampal neurons. Ngn3 is transiently exported from the cell nucleus to the cytoplasm when neuronal polarity is initiated, suggesting that the nucleo-cytoplasmic transport of the protein is important for its action on neuronal development. In this study, we identified for the first time a functional nuclear export sequence (NES2; ¹³¹YIWALTQTLRIA¹⁴²) in Ngn3. The green fluorescent protein (EGFP)-NES2 fusion protein was localized in the cytoplasm and its nucleo-cytoplasmic shuttling was blocked by the CRM1 specific export inhibitor leptomycin B. Mutation of a leucine residue to alanine (L135A) in the NES2 motif resulted in both cytoplasmic and nuclear localization of the EGFP-NES2 fusion protein and in the nuclear accumulation of ectopic full-length myc-Ngn3. In addition, point mutation of the leucine 135 counteracted the effects of Ngn3 on neuronal morphology and synaptic inputs indicating that the cytoplasmic localization of Ngn3 is important for neuronal development. Pharmacological perturbation of the cytoskeleton revealed that cytoplasmic Ngn3 is associated with microtubules.


Seed Endophyte Microbiome of Crotalaria pumila Unpeeled: Identification of Plant-Beneficial Methylobacteria.
Metal contaminated soils are increasing worldwide. Metal-tolerant plants growing on metalliferous soils are fascinating genetic and microbial resources. Seeds can vertically transmit endophytic microorganisms that can assist next generations to cope with environmental stresses, through yet poorly understood mechanisms. The aims of this study were to identify the core seed endophyte microbiome of the pioneer metallophyte Crotalaria pumila throughout three generations, and to better understand the plant colonisation of the seed endophyte Methylobacterium sp. Cp3. Strain Cp3 was detected in C. pumila seeds across three successive generations and showed the most dominant community member. When inoculated in the soil at the time of flowering, strain Cp3 migrated from soil to seeds. Using confocal microscopy, Cp3-mCherry was demonstrated to colonise the root cortex cells and xylem vessels of the stem under metal stress. Moreover, strain Cp3 showed genetic and in planta potential to promote seed germination and seedling development. We revealed, for the first time, that the seed microbiome of a pioneer plant growing in its natural environment, and the colonisation behaviour of an important plant growth promoting systemic seed endophyte. Future characterization of seed microbiota will lead to a better understanding of their functional contribution and the potential use for seed-fortification applications.


Automatic Machine-Learning-Based Outcome Prediction in Patients With Primary Intracerebral Hemorrhage.
Background: A predictive model can provide physicians, relatives, and patients the accurate information regarding the severity of disease and its predicted outcome. In this study, we used an automated machine-learning-based approach to construct a prognostic model to predict the functional outcome in patients with primary intracerebral hemorrhage (ICH). Methods: We retrospectively collected data on demographic characteristics, laboratory studies and imaging findings of 333 patients with primary ICH. The functional outcomes at the 1st and 6th months after ICH were defined by the modified Rankin scale. All of the attributes were used for preprocessing and for automatic model selection with Automatic Waikato Environment for Knowledge Analysis. Confusion matrix and areas under the receiver operating characteristic curves (AUC) were used to test the predictive performance. Results: Among the models tested, the random forest provided the best predictive performance for functional outcome. The overall accuracy for predicting the 1st month outcome was 83.1%, with 77.4% sensitivity and 86.9% specificity, and the AUC was 0.899. The overall accuracy for predicting the 6th month outcome was 83.9%, with 72.5% sensitivity and 90.6% specificity, and the AUC was 0.917. Conclusions: Using an automatic machine learning technique to predict functional outcome after ICH is feasible, and the random forest model provides the best predictive performance across all tested models. This prediction model may provide information regarding functional outcome for clinicians that will help provide appropriate medical care for patients and information for their caregivers.


Mobyle: a new full web bioinformatics framework.
For the biologist, running bioinformatics analyses involves a time-consuming management of data and tools. Users need support to organize their work, retrieve parameters and reproduce their analyses. They also need to be able to combine their analytic tools using a safe data flow software mechanism. Finally, given that scientific tools can be difficult to install, it is particularly helpful for biologists to be able to use these tools through a web user interface. However, providing a web interface for a set of tools raises the problem that a single web portal cannot offer all the existing and possible services: it is the user, again, who has to cope with data copy among a number of different services. A framework enabling portal administrators to build a network of cooperating services would therefore clearly be beneficial. We have designed a system, Mobyle, to provide a flexible and usable Web environment for defining and running bioinformatics analyses. It embeds simple yet powerful data management features that allow the user to reproduce analyses and to combine tools using a hierarchical typing system. Mobyle offers invocation of services distributed over remote Mobyle servers, thus enabling a federated network of curated bioinformatics portals without the user having to learn complex concepts or to install sophisticated software. While being focused on the end user, the Mobyle system also addresses the need, for the bioinfomatician, to automate remote services execution: PlayMOBY is a companion tool that automates the publication of BioMOBY web services, using Mobyle program definitions. The Mobyle system is distributed under the terms of the GNU GPLv2 on the project web site (http://bioweb2.pasteur.fr/projects/mobyle/). It is already deployed on three servers: http://mobyle.pasteur.fr, http://mobyle.rpbs.univ-paris-diderot.fr and http://lipm-bioinfo.toulouse.inra.fr/Mobyle. The PlayMOBY companion is distributed under the terms of the CeCILL license, and is available at http://lipm-bioinfo.toulouse.inra.fr/biomoby/PlayMOBY/.


The mouse thymidylate synthase promoter: essential elements are in close proximity to the transcriptional initiation sites.
The promoter region of the mouse thymidylate synthase gene was analyzed by deletion and site-directed mutagenesis. Elimination of an upstream Sp1 element reduced expression threefold, whereas elimination of an adenovirus upstream stimulatory factor element had little effect. All of the upstream elements that are essential for promoter activity are located within 22 nucleotides of the first transcriptional initiation site.


Enzymatic hydrolysis of soy protein for nutritional fortification of low pH food.
Enzymatic hydrolysis of proteins is an attractive means of obtaining the changes in the functional properties often necessary for incorporating the protein in low pH food, e.g. soft drinks. The main problem associated with protein hydrolysis is the welldel-known formation of bitter-tasting peptides. The bitterness seems to be related to a too high degree of hydrolysis (DH). DH is defined as the percentage of cleaved peptide bonds and serves as the controlling parameter for the hydrolysis process. The choice of hydrolysis parameters is briefly discussed in relation to studies on the hydrolysis kinetics which in many respects can be described by classical enzyme kinetics. Several pilot plant productions of isoelectric soluble soy protein hydrolyzates have been carried out using soy isolate or soy concentrate as the raw material. The preferred enzyme for this type of hydrolysis is Alcalase 0.6 L, a food-grade liquid preparation of subtilism Carlsberg. The chosen hydrolysis parameters are : Substrate conc. : 8 per cent protein (N x 6.25). Enzyme-substrate ratio : 2 per cent. pH : 8.0. Temperature : 50 degrees C. The optimal DH value with regard to taste seems to be 10 per cent, corresponding to about 2 hours' hydrolysis time. The hydrolysis is terminated by the addition of citric or malic acids until pH reaches a value of 4.2. Separation of the supernatant from the unconverted protein is carried out in a continuous, solids-ejecting centrifuge. Preliminary rat trials show a satisfactory nutritional quality of the hydrolyzates. Possible end-uses for soy protein hydrolyzates are briefly discussed.


The study between the dynamics and the X-ray anatomy and regularizing effect of gallbladder on bile duct sphincter of the dog.
To study the relationship between the radiological anatomy and the dynamics on bile duct sphincter in bile draining and regularizing effect of gallbladder. Sixteen healthy dogs weighing 18 kg to 25 kg were divided randomly into control group and experimental group (cholecystectomy group). Cineradiography, manometry with perfusion, to effect of endogenous cholecystokinin and change of ultrastructure were employed. According to finding of the choledochography and manometry, in control group the intraluminal basal pressure of cephalic cyclic smooth muscle of choledochal sphincter cCS was 9.0+/-2.0 mmHg and that of middle oblique smooth muscle of choledochal sphincter (mOS) was 16.8+/-0.5 mmHg, the intraluminal basal pressure of cCS segment was obviously lower than that of mOS (P<0.01) in the interval period of bile draining, but significative difference of intraluminal basal pressure of the mOS segment was not found between the interval period of bile draining (16.8+/-0.5 mmHg) and the bile flowing period (15.9+/-0.9 mmHg) (P>0.05). The motility of cCS was mainly characterized by rhythmically concentric contraction, just as motility of cCS bile juice was pumped into the mOS segment in control group. And motility of mOS segment showed mainly diastolic and systolic activity of autonomically longitudinal peristalsis. There was spasmodic state in cCS and mOS segment and reaction to endogenous cholecystokinin was debased after cholecystectomy. The change of ultrastructure of cCS portion showed mainly that the myofilaments of cell line in derangement and mitochondria is swelling. During fasting, the cCS portion has a function as similar cardiac "pump" and it is main primary power source in bile draining, and mOS segment serves mainly as secondary power in bile draining. The existence of the intact gallbladder is one of the important factors in guaranteeing the functional coordination between the cCS and mOS of bile duct sphincter. There is dysfunction in the cCS and mOS with cholecystectomy.


Molecular recognition of a lipophilic guanosine derivative in Langmuir films at the air-water interface.
Molecular recognition of a lipophilic deoxyguanosine derivative at the air-water interface was investigated by film balance experiments and Brewster Angle Microscopy. Results showed that guanosine, despite strong tendency towards self-assembly, interacts with both complementary and noncomplementary liponucleosides (lipophilic derivatives of deoxycytidine and deoxythymidine). At surface pressures below 17mN/m, attractive and repulsive interactions were present in case of both mixed monolayers and were the strongest at guanosine molar fractions of 0.5 and 0.75. At higher values of surface pressure, deoxyguanosine-deoxycytidine interactions were strictly attractive and were present only for monolayers with guanosine molar fraction of 0.75. On the contrary, attractive and repulsive interactions remained present in case of deoxyguanosine-deoxythymidine mixed monolayers. This indicates that interactions between guanosine and cytidine are much stronger than guanosine-thymidine interactions. Interactions for none of the nucleoside pairs, however, are specific and π-stacking interactions between the aromatic planes of liponucleoside derivatives probably dominate over hydrogen bonding interactions. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio.


Iron-Based Metal-Organic Frameworks as Platform for H2S Selective Conversion: Structure-Dependent Desulfurization Activity.
Three classical Fe-MOFs, viz., MIL-100(Fe), MIL-101(Fe), and MIL-53(Fe), were synthesized to serve as platforms for the investigation of structure-activity relationship and catalytic mechanism in the selective conversion of H2S to sulfur. The physicochemical properties of the Fe-MOFs were characterized by various techniques. It was disclosed that the desulfurization performances of Fe-MOFs with well-defined microstructures are obviously different. Among these, MIL-100(Fe) exhibits the highest catalytic performance (ca. 100% H2S conversion and 100% S selectivity at 100-180 °C) that is superior to that of commercial Fe2O3. Furthermore, the results of systematic characterization and DFT calculation reveal that the difference in catalytic performance is mainly because of discrepancy in the amount of Lewis acid sites. A plausible catalytic mechanism has been proposed for H2S selective conversion over Fe-MOFs. This work provides critical insights that are helpful for rational design of desulfurization catalysts.


Mapping value based planning and extensively trained choice in the human brain.
Investigations of the underlying mechanisms of choice in humans have focused on learning from prediction errors, leaving the computational structure of value based planning comparatively underexplored. Using behavioral and neuroimaging analyses of a minimax decision task, we found that the computational processes underlying forward planning are expressed in the anterior caudate nucleus as values of individual branching steps in a decision tree. In contrast, values represented in the putamen pertain solely to values learned during extensive training. During actual choice, both striatal areas showed a functional coupling to ventromedial prefrontal cortex, consistent with this region acting as a value comparator. Our findings point toward an architecture of choice in which segregated value systems operate in parallel in the striatum for planning and extensively trained choices, with medial prefrontal cortex integrating their outputs.


Drop penetration into porous powder beds.
The kinetics of drop penetration were studied by filming single drops of several different fluids (water, PEG200, PEG600, and HPC solutions) as they penetrated into loosely packed beds of glass ballotini, lactose, zinc oxide, and titanium dioxide powders. Measured times ranged from 0.45 to 126 s and depended on the powder particle size, viscosity, surface tensions, and contact angle. The experimental drop penetration times were compared to existing theoretical predictions by M. Denesuk et al. (J. Colloid Interface Sci.158, 114, 1993) and S. Middleman ("Modeling Axisymmetric Flows: Dynamics of Films, Jets, and Drops," Academic Press, San Diego, 1995) but did not agree. Loosely packed powder beds tend to have a heterogeneous bed structure containing large macrovoids which do not participate in liquid flow but are included implicitly in the existing approach to estimating powder pore size. A new two-phase model was proposed where the total volume of the macrovoids was assumed to be the difference between the bed porosity and the tap porosity. A new parameter, the effective porosity epsilon(eff), was defined as the tap porosity multiplied by the fraction of pores that terminate at a macrovoid and are effectively blocked pores. The improved drop penetration model was much more successful at estimating the drop penetration time on all powders and the predicted times were generally within an order of magnitude of the experimental results.


[The application of stem cells, visible and infrared light in regenerative medicine. Part 1].
The present article was designed to overview the experimental studies of visible and infrared light irradiation of human and animal stem cells (SC) in vitro and in vivo for the evaluation of its photobiomodulatory effects. The results will be used to elaborate substantiation for the choice of the parameters of SC light irradiation and to develop recommendations for the application of this method in regenerative medicine (RM). The clinical application of light irradiation is a matter of contrsy, in the first place due to the difficulties encountered in the rational choice of irradiation parameters. The theoretical substantiation of such choice remains a stumbling block too despite the long history of photoghromotherapy. There is thus far no reliable theoretical basis for the adequate choice of such irradiation parameters as power density, radiation dose, and exposure time. The experiences with the light application for the purpose of regenerative medicine have never been summarized. The present review encompasses 78 articles selected for the basic analysis that report the studies with the use of a variety of SC types. The analysis has demonstrated that clinical investigations into the influence of light on the stem cells are still in their infancy. It was shown that the irradiation parameters need to be chosen taking into consideration the type of the stem cells. Different authors report the achievement of the maximum SC proliferation and differentiation rates at energy densities as high as 50 mW/sq.cm, small radiation doses (around 1 J/sq.cm) and exposure time (on the order of seconds). The general conclusion for Parts 1 and II of this communication will be presented in the next issue of this journal (number 2, 2015).


Postnatal changes in pulmonary mechanics and energetics of infants with respiratory distress syndrome following surfactant treatment.
Postnatal alterations in pulmonary mechanics, energetics and functional residual capacity (FRC) describe the structural maturation of the preterm respiratory system. To evaluate longitudinal changes in pulmonary function in infants with respiratory distress syndrome (RDS) treated with oxygen, positive pressure ventilation and synthetic surfactant (Exosurf). Serial pulmonary function tests were performed in surfactant-treated infants [mean +/- SD birth weight (BW) = 1,112 +/- 276 g, gestational age (GA) = 29 +/- 3 weeks] at postnatal ages: <3 days, 1, 2, 3, 4 and 6-8 weeks until term postmenstrual age (PMA). Tidal volume, pulmonary compliance (C(L)), pulmonary resistance (R(T)) and flow-resistive work were analyzed following simultaneous measurements of airflow and transpulmonary pressure signals. Serial FRC measurements were made in a randomly selected group. Prior to 28 weeks' PMA, C(L) was unchanged irrespective of GA. At age 1 week the likelihood ratio (LR) for bronchopulmonary dysplasia (BPD) based on C(L), R(T) and GA was predicted to be >90% for those with BW <750 g (LR >100) as compared to <10% probability (LR = 0.3) for infants >1,500 g. Significant linear increase in C(L) to PMA was evident >28 weeks' PMA (r = 0.86, p < 0.01) at 0.17 ml/cm H2O/kg/week. By term PMA, mean C(L) was 2.60 +/- 0.07 ml/cm H2O. Improvements in FRC of preterm infants with RDS who recovered occur at a more rapid rate ( approximately 25 ml/kg) compared to those who developed BPD ( approximately 20 ml/kg). Slow but incremental postnatal pulmonary improvement, minimal <28 weeks' PMA, were comparable for all infants. Along with diminished FRC, these changes reflect persistent deleterious effects of positive pressure ventilation, alveolar hyperoxia and unrecognized pulmonary overdistension.


Genotype shift in human coronavirus OC43 and emergence of a novel genotype by natural recombination.
Human coronavirus (HCoV) OC43 is the most prevalent HCoV in respiratory tract infections. Its molecular epidemiological characterization, particularly the genotyping, was poorly addressed. The full-length spike (S), RNA-dependent RNA polymerase (RdRp), and nucleocapsid (N) genes were amplified from each respiratory sample collected from 65 HCoV-OC43-positive patients between 2005 and 2012. Genotypes were determined by phylogenetic analysis. Recombination was analyzed based on full-length viral genome sequences. Clinical manifestations of each HCoV genotype infection were compared by reviewing clinical records. Sixty of these 65 samples belong to genotypes B, C and D. The remaining five strains had incongruent positions in the phylogenetic trees of the S, RdRp and N genes, suggesting a novel genotype emerging, designated as genotype E. Whole genome sequencing and bootscan analysis indicated that genotype E is generated by recombination between genotypes B, C and D. Temporal analysis revealed a sequential genotype replacement of C, B, D and E over the study period with genotype D being the dominant genotype since 2007. The novel genotype E was only detected in children younger than three years suffering from lower respiratory tract infections. Our results suggest that HCoV-OC43 genotypes are evolving. Such genotype shift may be an adapting mechanism for HCoV-OC43 maintaining its epidemic.


Tools of the trade: MicroCT reveals native structure and functional morphology of organs that drive caterpillar-ant interactions.
Caterpillars of many lycaenid butterflies are tended by ants that offer protection from predators and parasitoids. Specialized structures such as glands, ciliary organs and chitinous ornamentation in caterpillars play key roles in the underlying tactile, acoustic, and chemical communication between caterpillars and ants. Although the ecological, evolutionary, and behavioural aspects of these interactions are well studied, the mechanisms (i.e., the functional morphology) that drive the specialized interactive organs are poorly characterized. We used advanced X-ray microtomography (MicroCT) to delineate internal, native morphology of specialized larval dew patches, nectar glands, and tactile ciliary organs that mediate interactions between Crematogaster ants and caterpillars of the obligate myrmecophilous Apharitis lilacinus butterfly. Our non-destructive MicroCT analysis provided novel 3-D insights into the native structure and positions of these specialized organs in unmatched detail. This analysis also suggested a functional relationship between organ structures and surrounding muscles and nervation that operate the glands and tactile organs, including a 'lasso bag' control mechanism for dew patches and muscle control for other organs. This provided a holistic understanding of the organs that drive very close caterpillar-ant interactions. Our MicroCT analysis opens a door for similar structural and functional analysis of adaptive insect morphology.


Assessing Treatment of Floating Lateral Mass (FLM) Fractures of the Subaxial Cervical Spine.
Retrospective cohort. The purpose of the study was to evaluate differences across surgical approaches (anterior, posterior, or combined anterior-posterior) in terms of outcomes following treatment for floating lateral mass (FLM) fractures. Furthermore, we sought to determine whether operative approach to FLM fracture treatment remains superior to nonoperative treatment in terms of clinical outcomes. FLM fractures of the subaxial cervical spine involves separation of the lateral mass from the vertebrae via a disruption of both the lamina and pedicle, resulting in a disconnection of the superior and inferior articular processes. This subset of cervical spine fractures is highly unstable, making proper treatment selection of great importance. In this single-center, retrospective study, we identified patients meeting the definition of an FLM fracture. Radiological imaging from the date of injury was reviewed to ensure presence this injury pattern. Treatment course was assessed to determine nonoperative versus operative treatment. Operative treatment was divided into patients who underwent anterior, posterior, or combined anterior-posterior spinal fusion. We then reviewed postoperative complications among each of the subgroups. Forty-five patients were determined to have a FLM fracture over a 10-year span. The nonoperative group had n=25, and evidently, there were no patients that crossed over to surgery due to subluxation of the cervical spine after nonoperative treatment. The operative treatment group had n=20, and consisted of 6 anterior, 12 posterior, and 2 combined approaches. Complications appeared in posterior and combined groups. Two hardware failures were noted in the posterior group, along with two postoperative respiratory complications in the combined group. No complications were observed for the anterior group. None of the nonoperative patients in this study required further operation or management of their injury, indicating nonoperative treatment as a potentially satisfactory management for appropriately selected FLM fractures.


Intracranial oligodendrogliomas: imaging findings in 35 untreated cases.
The radiographic findings in 35 cases of untreated intracranial oligodendrogliomas were reviewed. The mean age of the patients was 34.6 years, and seizure disorder and headache were the most frequent presenting symptoms. Slightly less than two-thirds of the tumors were histologically pure and almost half were low-grade. Most lesions were cerebral and peripheral in location, and the majority were in the frontal lobes. On CT the tumors were usually hypo- or isodense. Contrast enhancement of tumor occurred in nearly half the cases, and was usually mild and poorly defined. Tumor calcification often occurred, and hemorrhage or cystic formation was not infrequent. Occasionally, calvarial erosion was associated with the tumors because of their peripheral location and slow-growing nature. The lesions were usually sharply demarcated and without edema. MR most frequently revealed hypointense lesions on T1-weighted images and abnormal hyperintensity on T2-weighted scans. In regard to grading or purity of oligodendrogliomas, no significant correlations were found except for a suggestion that higher-grade and mixed tumors tend to enhance more often on CT. The radiographic features of oligodendroglioma are quite characteristic but not pathognomic. A high preoperative suspicion might lead to more appropriate tumor management. MR, although less sensitive in detecting tumor calcification, is superior to CT in defining the tumor extent, which is beneficial for surgical and postsurgical radiotherapy planning.


Isolation of neuropeptide-containing vesicles from the guinea pig ileum.
Three distinct vesicle fractions enriched 40-60 times in the neuropeptides substance P, somatostatin, and vasoactive intestinal peptide (VIP) were prepared from the myenteric plexus of guinea pig ileum by density gradient centrifugation in a small zonal rotor. Mean densities (in g X ml-1) and diameters (in nm) of the three classes of vesicles were: substance P, 1.123, 65; somatostatin, 1.138, 37; VIP, 1.148, 110; standard deviations were about 5%. These peaks were distinct from the peak of acetylcholine-containing vesicles of density 1.066 g X ml-1 and diameter 61 nm. When a relatively mild method of homogenization was used a second peak of acetylcholine appeared in the same region of the gradient as VIP and the VIP was larger. This may represent a class of vesicles containing both acetylcholine and VIP, though cosedimentation of two classes of vesicles of almost the same density and similar fragility, one containing VIP and the other acetylcholine, cannot be entirely excluded on present evidence.