Neutrophil recruitment by fetal porcine endothelial cells: implications in scarless fetal wound healing.
Fetal dermal wounds heal scarlessly and with a minimal inflammatory response. When a robust inflammatory response is induced at the site of fetal dermal wounds by the application of cytokines, healing results in fibrosis. To test the hypothesis that the reduced inflammatory response in fetal wounds is due to impaired fetal leukocyte-endothelial interactions, the contributions of fetal endothelial cells to the inflammatory response in the fetus were investigated. Endothelial cells isolated from blood vessels of adult and mid-gestational fetal pigs were cultured in media until confluent monolayers were established. Adult porcine neutrophils were isolated and resuspended at a concentration of 1 million cells/mL. Interactions between neutrophils and endothelial cells were observed under static and flow conditions. Endothelial monolayers were exposed to neutrophils with and without prior stimulation of the endothelial cells with tumor necrosis factor alpha (TNF-alpha) for 4 h. The neutrophil-endothelial interactions were observed and analyzed for neutrophil adhesion, rolling velocity, and transmigration. Endothelial P-selectin mRNA expression was determined by real-time polymerase chain reaction (PCR). A novel in vitro model of fetal inflammation is described. Both adult and fetal endothelial cells demonstrated a dose-dependent increase in neutrophil adhesion and transmigration with increasing doses of TNF-alpha. The fetal response was significantly lower than the adult. As expected, rolling velocity was lower at higher cytokine concentrations and had an inverse correlation with P-selectin mRNA expression. Fetal endothelial cells are less permissive to adhesion and transmigration of neutrophils than adult endothelial cells. This may contribute to the paucity of inflammation seen in the fetal response to dermal injury.


[Profile of the antimigraine drug almotriptan].
Triptans are the most relevant advance in the symptomatic treatment of migraine in the last decade. In this paper the basic properties as well as the efficacy and tolerability data of the last launched triptan, almotriptan, are reviewed. Almotriptan exhibits the highest bioavailability among triptans, this absorption being unaffected by sex, the presence of food in the stomach or by a migraine attack. At the recommended dose of 12.5 mg, almotriptan shows a rapid onset of effect, at 30 minutes, with efficacy parameters comparable to those of placebo. The potential advantages of almotriptan seem to be three: recurrence rate in the low range, lack of interactions with other drugs and a placebo-like tolerability profile, with a very low rate of chest symptoms. This balanced profile between efficacy and tolerability makes almotriptan 12.5 mg a good option for the symptomatic treatment of migraine.


Presence of the JAK2 V617F mutation in a patient with chronic neutrophilic leukemia and effective response to interferon α-2b.
Chronic neutrophilic leukemia (CNL) is a rare type of leukemia characterized by a proliferation mainly of mature neutrophils, elevated neutrophil-alkaline phosphatase activity, and no presence of the Philadelphia chromosome. The prognosis is generally poor and there is no consensus therapeutic strategy for the treatment of this disease. The JAK2 V617F mutation has been detected in patients with classical myeloproliferative disorders (MPD) including polycythemia vera and essential thrombocythemia and idiopathic myelofibrosis. In contrast, this same mutation has been detected in only 4 patients with CNL to date, suggesting that the JAK2 V617F mutation is a rare event in patients with atypical MPD. Here, we report a case of CNL with presence of the JAK2 V617F mutation. After treatment with interferon alfa-2b with 3 million units every other day for 1 month, the patient's white blood cell count was well controlled below 10.0 ×10⁹/l. At present, our patient remains symptomatically well and is maintained on interferon alfa-2b (3 million units twice a week), and his neutrophil count now averages around 8.0-10.0 ×10⁹/l.


Microbial monitoring of spacecraft and associated environments.
Rapid microbial monitoring technologies are invaluable in assessing contamination of spacecraft and associated environments. Universal and widespread elements of microbial structure and chemistry are logical targets for assessing microbial burden. Several biomarkers such as ATP, LPS, and DNA (ribosomal or spore-specific), were targeted to quantify either total bioburden or specific types of microbial contamination. The findings of these assays were compared with conventional, culture-dependent methods. This review evaluates the applicability and efficacy of some of these methods in monitoring the microbial burden of spacecraft and associated environments. Samples were collected from the surfaces of spacecraft, from surfaces of assembly facilities, and from drinking water reservoirs aboard the International Space Station (ISS). Culture-dependent techniques found species of Bacillus to be dominant on these surfaces. In contrast, rapid, culture-independent techniques revealed the presence of many Gram-positive and Gram-negative microorganisms, as well as actinomycetes and fungi. These included both cultivable and noncultivable microbes, findings further confirmed by DNA-based microbial detection techniques. Although the ISS drinking water was devoid of cultivable microbes, molecular-based techniques retrieved DNA sequences of numerous opportunistic pathogens. Each of the methods tested in this study has its advantages, and by coupling two or more of these techniques even more reliable information as to microbial burden is rapidly obtained.


[Combined activity of meliacin and foscarnet against different strains of herpes simplex virus type 1 using a three-dimensional model].
We evaluated the in vitro antiviral activity of meliacin combined with foscarnet on the herpes simplex type 1 (HSV-1) strains F and B2006 (tk-) replication. The effective concentrations for 50% inhibition of HSV-1 (F) were 12.5 micrograms/ml for meliacin and 15.7 micrograms/ml for foscarnet, while for HSV-1 (B2006) were 3.1 micrograms/ml and 126 micrograms/ml, respectively. The data were analyzed for quantitation of synergism, additivity, and antagonism of multiple drug effect by the three-dimensional model. Some of the meliacin -foscarnet combinations synergistically inhibited HSV-1 (F) and HSV-1 (B2006) replication in vitro at concentrations that did not reduce cellular viability.


A double-blind, randomized, placebo-controlled trial of augmentation with lamotrigine or placebo in patients concomitantly treated with fluoxetine for resistant major depressive episodes.
Evidence of the antidepressant efficacy of lamotrigine is increasing, although there are no placebo-controlled trials of lamotrigine augmentation in depression. The aim of this study was to assess if augmentation with lamotrigine was superior to placebo in patients who were receiving fluoxetine for resistant major depressive episodes. Twenty-three patients who had experienced at least 1 major depressive episode that was resistant to at least 1 prior trial of antidepressant therapy were selected. These patients were treated with fluoxetine, 20 mg/day, and concomitantly randomly assigned to receive either lamotrigine (N = 13) or placebo (N = 10) for 6 weeks. The dose of lamotrigine was titrated upward from 25 mg/day to 100 mg/day. Patients suffering from bipolar II disorder (N = 8) or from major depressive disorder (N = 15) (DSM-IV criteria) were enrolled, resulting in heterogeneity of the sample. The primary outcome measure was Hamilton Rating Scale for Depression score. Data were collected from 2000-2001. Lamotrigine was statistically superior to placebo on the Clinical Global Impressions scale at endpoint, both in absolute terms (mean +/- SD Clinical Global Impressions-Severity of Illness scores: lamotrigine, 2.15 +/- 1.28; placebo, 3.40 +/- 1.17; p =.0308) and using a responder analysis, with response defined as a Clinical Global Impressions-Improvement score of 2 or less (lamotrigine, 84.62% [N = 11]; placebo, 30.00% [N = 3]; p =.013). The effect of lamotrigine on Clinical Global Impressions scale scores was seen in both major depressive disorder and bipolar II disorder. Lamotrigine, however, failed to separate statistically from placebo on the Hamilton Rating Scale for Depression and Montgomery-Asberg Depression Rating Scale. This failure to differentiate on a primary outcome measure is essentially a negative study result. This result is most likely an artifact of the small sample size used and the resultant limited power of the study. The results of this trial add to the literature suggesting potential efficacy of the antidepressant profile of lamotrigine. In addition, this study points to a possible role of lamotrigine as an augmentation agent in depression.


Prenylation of Rab8 GTPase by type I and type II geranylgeranyl transferases.
Rab GTPases are post-translationally modified by addition of geranylgeranyl moieties to carboxyl-terminal cysteine residues. For Rab proteins ending with xxCC xCxC and CCxx motifs this modification is catalysed by geranylgeranyltransferase type II (GGTaseII), and is entirely dependent on the Rab substrate being bound to Rab escort protein (REP). Several Rab proteins contain carboxyl-terminal CaaL prenylation motifs typical of members of the Rho family, which are modified in a REP-independent manner by geranylgeranyltransferase type I (GGTaseI). The present studies show that one such Rab protein (Rab8), which ends with a CVLL motif, is uniquely able to serve as a substrate for either REP/GGTaseII or GGTaseI in cell-free assays. The modification of Rab8 by GGTaseI did not require REP, indicating that a REP-induced conformational change is not essential for exposure of the Rab carboxyl-terminal cysteine prenylation site. To determine whether one enzyme plays a predominant role in Rab8 prenylation in vivo, the incorporation of [3H]mevalonate into Rab8 was measured in human embryonal kidney 293 cells under conditions where the activity of GGTaseI, but not GGTaseII, was blocked by the peptidomimetic inhibitor GGTI-298. The GGTaseI inhibitor did not prevent prenylation of either overexpressed Myc-tagged Rab8 or endogenous Rab8, whereas prenylation of a known GGTaseI substrate with the same carboxyl-terminal motif, Cdc42Hs, was completely blocked. To rule out the possibility that the apparent prenylation of Rab8 by GGTaseII occurs only when GGTaseI activity is eliminated, metabolic labelling studies were carried out in the absence of the GGTaseI inhibitor, using a REP-binding-deficient Rab8 construct (Y78D) that cannot serve as a substrate for GGTaseII, but is indistinguishable from wild-type Rab8 as a substrate for GGTaseI. Prenylation of the Y78D mutant was reduced by 60-70% in intact cells, consistent with the conclusion that the majority of Rab8 is prenylated by the REP/GGTaseII system in vivo.


Genome-wide SNP scan of pooled DNA reveals nonsense mutation in FGF20 in the scaleless line of featherless chickens.
Scaleless (sc/sc) chickens carry a single recessive mutation that causes a lack of almost all body feathers, as well as foot scales and spurs, due to a failure of skin patterning during embryogenesis. This spontaneous mutant line, first described in the 1950s, has been used extensively to explore the tissue interactions involved in ectodermal appendage formation in embryonic skin. Moreover, the trait is potentially useful in tropical agriculture due to the ability of featherless chickens to tolerate heat, which is at present a major constraint to efficient poultry meat production in hot climates. In the interests of enhancing our understanding of feather placode development, and to provide the poultry industry with a strategy to breed heat-tolerant meat-type chickens (broilers), we mapped and identified the sc mutation. Through a cost-effective and labour-efficient SNP array mapping approach using DNA from sc/sc and sc/+ blood sample pools, we map the sc trait to chromosome 4 and show that a nonsense mutation in FGF20 is completely associated with the sc/sc phenotype. This mutation, common to all sc/sc individuals and absent from wild type, is predicted to lead to loss of a highly conserved region of the FGF20 protein important for FGF signalling. In situ hybridisation and quantitative RT-PCR studies reveal that FGF20 is epidermally expressed during the early stages of feather placode patterning. In addition, we describe a dCAPS genotyping assay based on the mutation, developed to facilitate discrimination between wild type and sc alleles. This work represents the first loss of function genetic evidence supporting a role for FGF ligand signalling in feather development, and suggests FGF20 as a novel central player in the development of vertebrate skin appendages, including hair follicles and exocrine glands. In addition, this is to our knowledge the first report describing the use of the chicken SNP array to map genes based on genotyping of DNA samples from pooled whole blood. The identification of the sc mutation has important implications for the future breeding of this potentially useful trait for the poultry industry, and our genotyping assay can facilitate its rapid introgression into production lines.


The ALIGN Framework: A Parent-Informed Approach to Prognostic Communication for Infants With Neurologic Conditions.
Clinicians often communicate complex, uncertain, and distressing information about neurologic prognosis to parents of critically ill infants. Although communication tools have been developed in other disciplines and settings, none address the unique needs of the neonatal and pediatric neurology context. We aimed to develop a parent-informed framework to guide clinicians in communicating information about neurologic prognosis. Parents of infants with neurologic conditions in the intensive care unit were enrolled in a longitudinal study of shared decision-making from 2018 to 2020. Parents completed semistructured interviews following recorded family meetings with the health care team, at hospital discharge, and 6 months after discharge. All interviews targeted information about parent preferences for prognostic disclosure. We analyzed the data using a conventional content analysis approach. Two study team members independently coded all interview transcripts, and discrepancies were resolved in consensus. We used NVIVO 12 qualitative software to index and organize codes. Fifty-two parents of 37 infants completed 123 interviews. Parents were predominantly mothers (n = 37/52, 71%) with a median age of 31 (range 19-46) years. Half were Black (n = 26/52, 50%), and a minority reported Hispanic ethnicity (n = 2/52, 4%). Inductive analysis resulted in the emergence of 5 phases of prognostic communication (Approach, Learn, Inform, Give support, and Next steps: ALIGN): (1) Approach: parents appreciated receiving consistent information about their child's neurologic outcome from clinicians who knew their child well. (2) Learn: parents valued when clinicians asked them how they preferred receiving information and what they already knew about their child's outcome prior to information delivery. (3) Inform: parents valued honest, thorough, and balanced information that disclosed prognostic uncertainty and acknowledged room for hope. (4) Give support: parents valued empathic communication and appreciated clinicians who offered real-time emotional support. (5) Next steps: parents appreciated clinicians who connected them to resources, including peer support. The ALIGN framework offers a novel, parent-informed strategy to effectively communicate neurologic prognosis. Although ALIGN represents key elements of a conversation about prognosis, each clinician can adapt this framework to their own approach. Future work will assess the effectiveness of this framework on communication quality and prognostic understanding.


Novel metal-organic frameworks with high stability for selectively sensing nitroaromatics.
The stability and functionality of metal-organic frameworks (MOFs) are particularly important in considering their potential and/or practical use. Herein we report two novel aldehyde functional group-containing MOFs [Zn2(L)(BDC)2]·2DMF·MeOH·H2O (1) and [Co2(L)(BDC)2]·2DMF·2MeOH·H2O (2) (DMF = N,N-dimethylformamide, MeOH = methanol) achieved by reactions of mixed organic ligands of 2,4-bis[(pyridin-4-yl)methoxy]benzaldehyde (L) and 1,4-benzenedicarboxylic acid (H2BDC) with the corresponding metal salts under solvothermal conditions. Both 1 and 2 are 2-fold interpenetrated three-dimensional (3D) frameworks. Particularly, 1 exhibits excellent water and wide-range pH stability, and a large Qst value for carbon dioxide (CO2) adsorption. Furthermore, photoluminescence (PL) studies indicate that 1 could be considered as a potential luminescent probe for detection of nitroaromatics.


[Mechanism of activation of the external pathway of NADH oxidation in myocardial mitochondria during hemorrhagic shock].
The changes in the oxidative metabolism of rabbit myocardium in hemorrhagic shock were studied. The external pathway of NADH oxidation was shown to be activated, while the rates of glutamate oxidation in the presence of an uncoupler were diminished as compared to control values. These changes are probably due to the activation of phospholipase A2 and lipid peroxidation, since their inhibitors eliminate with the same efficiency the activation of the external pathway of NADH oxidation both in normal and damaged myocardium.


Fluorogen activating protein-affibody probes: modular, no-wash measurement of epidermal growth factor receptors.
Fluorescence is essential for dynamic live cell imaging, and affinity reagents are required for quantification of endogenous proteins. Various fluorescent dyes can report on different aspects of biological trafficking, but must be independently conjugated to affinity reagents and characterized for specific biological readouts. Here we present the characterization of a new modular platform for small anti-EGFR affinity probes for studying rapid changes in receptor pools. A protein domain (FAP dL5**) that binds to malachite-green (MG) derivatives for fluorescence activation was expressed as a recombinant fusion to one or two copies of the compact EGFR binding affibody ZEGFR:1907. This is a recombinant and fluorogenic labeling reagent for native EGFR molecules. In vitro fluorescence assays demonstrated that the binding of these dyes to the FAP-affibody fusions produced thousand-fold fluorescence enhancements, with high binding affinity and fast association rates. Flow cytometry assays and fluorescence microscopy demonstrated that these probes label endogenous EGFR on A431 cells without disruption of EGFR function, and low nanomolar surface Kd values were observed with the double-ZEGFR:1907 constructs. The application of light-harvesting fluorogens (dyedrons) significantly improved the detected fluorescence signal. Altering the order of addition of the ligand, probe, and dyes allowed differentiation between surface and endocytotic pools of receptors to reveal the rapid dynamics of endocytic trafficking. Therefore, FAP/affibody coupling provides a new approach to construct compact and modular affinity probes that label endogenous proteins on living cells and can be used for studying rapid changes in receptor pools involved in trafficking.


Ca2+ efflux mechanisms following depolarization evoked calcium transients in cultured rat sensory neurones.
1. We have used a combination of microfluorimetry and patch-clamp techniques to investigate cytoplasmic Ca2+ ([Ca2+]i) buffering in response to physiological Ca2+ loads in neurones cultured from the dorsal root ganglia of the rat. 2. In cells loaded with Indo-1 AM and using high resistance microelectrodes to initiate and record action potentials, single action potentials were associated with a measurable rise in [Ca2+]i. Short trains of action potentials evoked [Ca2+]i transients with monoexponential recovery rates with time constants of around 5 s. 3. Similar Ca2+ buffering properties were seen in cells perfused with patch-clamp pipettes in the whole-cell recording mode suggesting that the slow (seconds) Ca2+ buffering properties were not seriously perturbed by the recording technique. 4. In cells held under voltage clamp, reversal of the Na(+)-Ca2+ exchanger driving force had a small but significant effect on the rate of Ca2+ removal. 5. Increasing extracellular pH or adding vanadate (200 microM) to the internal solution dramatically slowed the rate of recovery. Addition of calmidazolium to the pipette solution also produced a significant but much less dramatic slowing of Ca2+ efflux. 6. The results demonstrate that the activity of a plasmalemmal Ca(2+)-ATPase is important for the removal of somatic Ca2+ loads of a similar amplitude to those generated by the firing of a few action potentials.


Food reward-induced neurotransmitter changes in cognitive brain regions.
Recent evidence indicates that mechanisms involved in reward and mechanisms involved in learning interact, in that reward includes learning processes and learning includes reward processes. In spite of such interactions, reward and learning represent distinct functions. In the present study, as part of an examination of the differences in learning and reward mechanisms, it was assumed that food principally affects reward mechanisms. After a brief period of fasting, we assayed the release of three neurotransmitters and their associated metabolites in eight brain areas associated with learning and memory as a response to feeding. Using microdialysis for the assay, we found changes in the hippocampus, cortex, amygdala, and the thalamic nucleus, (considered cognitive areas), in addition to those in the nucleus accumbens and ventral tegmental area (considered reward areas). Extracellular dopamine levels increased in the nucleus accumbens, ventral tegmental area, amygdala, and thalamic nucleus, while they decreased in the hippocampus and prefrontal cortex. Dopamine metabolites increased in all areas tested (except the dorsal hippocampus); changes in norepinephrine varied with decreases in the accumbens, dorsal hippocampus, amygdala, and thalamic nucleus, and increases in the prefrontal cortex; serotonin levels decreased in all the areas tested; although its metabolite 5HIAA increased in two regions (the medial temporal cortex, and thalamic nucleus). Our assays indicate that in reward activities such as feeding, in addition to areas usually associated with reward such as the mesolimbic dopamine system, other areas associated with cognition also participate. Results also indicate that several transmitter systems play a part, with several neurotransmitters and several receptors involved in the response to food in a number of brain structures, and the changes in transmitter levels may be affected by metabolism and transport in addition to changes in release in a regionally heterogeneous manner. Food reward represents a complex pattern of changes in the brain that involve cognitive processes. Although food reward elements overlap with other reward systems sharing some neurotransmitter compounds, it significantly differs indicating a specific reward to process for food consumption. Like in other rewards, both learning and cognitive areas play a significant part in food reward.


Roles and Responsibilities in the Transition to Working Independently: A Qualitative Study of Recently Graduated Radiographers' Perspectives in Australia.
Medical imaging features along the entire healthcare continuum and is known for its fast-paced technological evolution which enables it to keep up with the demands of the healthcare system to provide safe, quality services. The overall efficacy and efficiency of the system depends on practitioners' clinical competence, achieved through professional education and continuous professional development. Recent studies have revealed concerns regarding newly graduated healthcare professionals' preparedness and readiness to handle actual practice. We conducted qualitative face-to-face and telephonic interviews with a convenient and purposive sample of 23 participants consisting of recently graduated radiographers (n=14), radiography students (n=5) and supervising radiographers (n=4) in Australia. Verbatim transcriptions were analyzed inductively to identify themes pertaining to perspectives and experiences of the work readiness of novice radiographers. The findings of our study suggest that the workplace immersion and transitioning of recently graduated radiographers into their professional roles requires a process of experiential learning and honing of knowledge and skills if they are to function efficiently and independently in a team-oriented workplace. Radiographic services are spread across various levels of care and are an integral part of the organizational structure of a healthcare system. Maladaptive transitions to the workplace may be the result of low self-confidence, a lack of support, uncertainty in inter-collegial interactions, or unrealistic performance expectations. The overarching themes of communication and interaction emerged clearly as recently graduated radiographers navigated the four roles of coordinator, collaborator, mediator, and advocate. The application of radiographic skills is embedded in a workplace culture of communication and safety. Transitioning to independent practice takes place in a complex, multifaceted environment and is accompanied by internal and external expectations. Because each workplace has a unique context, system and culture, no novice radiographic professional can ever be fully prepared through pre-service training and workplace induction.


The structural properties and antigenicity of soybean glycinin by glycation with xylose.
Soybean glycinin is considered a major allergenic protein, and glycation is widely used to reduce the allergenic potential of present allergens. Glycation of soybean glycinin with xylose at 55 °C for different lengths of time was investigated. The extent of Maillard reaction was reflected through the content changing of free amino groups, color analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Alteration in the structural properties of glycinin was characterized by Fourier transform infrared spectroscopy, and antigenicity was evaluated by indirect competitive enzyme-linked immunosorbent assay. The changes in the color of glycinin-xylose samples and the reduction of free amino group content in proteins indicated that the Maillard reaction occurred. The degree of glycation increased in glycated samples with the increase in reaction time. Glycation induced the changes in the secondary structure of glycinin and the ordered structure of proteins increased during the glycation reaction. The antigenicity of glycinin was reduced with the increase in reaction time. After glycation for 12 h, the antigenicity of glycinin declined about 18% compared with native glycinin. The application of glycation may be an efficient method to reduce the antigenicity of soybean glycinin. © 2016 Society of Chemical Industry.


Trans-complementation of the C gene of human and the P gene of woodchuck hepadnaviruses.
A 5 bp insertion was introduced into the BstEII site at nucleotide 2815 in DNA of hepatitis B virus (HBV) and a mutant HBV genome was produced, which coded for envelope and core proteins, but not for DNA polymerase, due to a frameshift. Cultured hepatoma cells (HepG2) were simultaneously transfected with a plasmid harbouring a tandem dimer of the mutant HBV DNA and another plasmid harbouring a tandem dimer of DNA of woodchuck hepatitis virus or duck hepatitis B virus. The replication of mutant HBV DNA, incapable of encoding DNA polymerase, was accomplished by cotransfecting woodchuck hepatitis virus DNA, but not by duck hepatitis B virus DNA. These results indicated a trans-complementation of the C and P genes in mammalian hepadnaviruses beyond a species barrier.


Elevated factor Xa activity in the blood of asymptomatic patients with congenital antithrombin deficiency.
The presence of congenital antithrombin deficiency has been consistently shown to predispose patients to venous thrombosis. We have utilized the prothrombin fragment F1+2 radioimmunoassay to quantitate factor Xa activity in the blood of 22 asymptomatic individuals with this clinical disorder not receiving antithrombotic therapy. The mean level of F1+2 was significantly elevated in these patients as compared to normal controls (3.91 vs. 1.97 nM, P less than 0.001). The metabolic behavior of 131 I-F1+2 was found to be similar in antithrombin-deficient subjects and normal individuals. The hemostatic system hyperactivity as measured by the F1+2 assay could be specifically corrected by raising the plasma antithrombin levels of the above asymptomatic individuals into the normal range. This study provides the first demonstration that the prethrombotic state can be biochemically defined as an imbalance between the production and inhibition of factor Xa enzymatic activity within the human circulation. It is known that antithrombin and alpha 1-proteinase inhibitor (PI) are the major inhibitors of factor Xa in human plasma in the absence of heparin. To further evaluate the mechanism by which antithrombin functions as an inhibitor of factor Xa in humans, we studied five patients who exhibited severe congenital deficiencies of alpha 1-PI. Our results indicated that the plasma of these subjects showed virtually identical decreases in plasma antifactor Xa activity in the absence of heparin when compared to antithrombin-deficient individuals, but the plasma F1+2 levels in the alpha 1-PI deficient population were not significantly different than normal. This data suggests that alpha 1-PI does not function as a major inhibitor of factor Xa in vivo, and that a tonically active heparin-dependent mechanism exists in humans for accelerating the neutralization of this enzyme by antithrombin.


The population health record: concepts, definition, design, and implementation.
In 1997, the American Medical Informatics Association proposed a US information strategy that included a population health record (PopHR). Despite subsequent progress on the conceptualization, development, and implementation of electronic health records and personal health records, minimal progress has occurred on the PopHR. Adapting International Organization for Standarization electronic health records standards, we define the PopHR as a repository of statistics, measures, and indicators regarding the state of and influences on the health of a defined population, in computer processable form, stored and transmitted securely, and accessible by multiple authorized users. The PopHR is based upon an explicit population health framework and a standardized logical information model. PopHR purpose and uses, content and content sources, functionalities, business objectives, information architecture, and system architecture are described. Barriers to implementation and enabling factors and a three-stage implementation strategy are delineated.


Range-of-motion restriction and craniofacial tissue-interface pressure from four cervical collars.
Occipital pressure ulcers are well-known complications for trauma patients wearing cervical collars. We assessed the effects of four commercial cervical collars (Aspen, Philadelphia, Miami J, and Miami J with Occian back [Miami J/Occian]) on cervical range of motion (CROM) and mandibular and occipital tissue-interface pressure (TIP). Forty-eight healthy volunteers (24 men, 24 women; mean age, 38.9 years +/- 10.5 years) were stratified by body mass index. CROM was measured in the seated position without and with collars. Sagittal, coronal, and rotatory CROM was measured with a goniometer. Occipital and mandibular pressures were mapped with subjects in upright and supine positions. All collars significantly restricted CROM in all planes (p < 0.001). The Philadelphia and standard Miami J collars were the most restrictive. The Aspen collar was the least restrictive for flexion and rotation. The Miami J/Occian back was the least restrictive for extension and lateral flexion. For supine measurements, Miami J and Miami J/Occian back had the lowest mean TIP, whereas Aspen and Philadelphia collars had the greatest (p < 0.001). For upright measurements, the Miami J/Occian back produced the smallest mean TIPs; the other collars, ranked by ascending TIP, were Philadelphia, Miami J, and Aspen (p < 0.001). Philadelphia and Miami J collars had significant collar-body mass index interaction effects on supine occiput mean pressure (p = 0.04). Miami J and Philadelphia collars restricted CROM to the greatest extent. Miami J and Miami J/Occian back had the lowest levels of mandibular and occipital pressure; these collars may markedly reduce the risk of occipital pressure ulcers without compromising immobilization.


PKCdelta-dependent deubiquitination and stabilization of Gadd45 in A431 cells overexposed to EGF.
Epidermal growth factor (EGF) receptor-overexpressing p53-deficient A431 cells response to toxic dose of EGF by G1 arrest and apoptosis was studied. We previously reported an increased expression of growth arrest and DNA-damage-inducible gene, Gadd45, in EGF-overexposed A431 cells. The mechanism for this induction was increased half-lives of mRNA and protein. In this study, using phorbol ester (a PKC activator) and specific inhibitors of PKC isoforms, we showed that protein kinase C-delta (PKCdelta) was involved in the increase of Gadd45 protein stability. We further demonstrated that Gadd45 is ubiquitinated and is regulated by proteolysis. While EGF induced ubiquitination of total cellular proteins, there was a decrease in Gadd45 ubiquitination, which could be inhibited by Rottlerin, a PKCdelta-specific inhibitor. These results suggest that an increase in Gadd45 stability may involve PKCdelta-dependent ubiquitin-proteasome pathway.


Finding unexpected patterns in microarray data.
We describe the performance of a protocol based on the sequential application of unsupervised and supervised methods to analyze microarray samples defined by a combination of factors. Correspondence analysis is used to visualize the emerging patterns of three set of novel or previously published data: photoreceptor mutants of Arabidopsis grown under different light/dark conditions, Arabidopsis exposed to different types of biotic and abiotic stress, and human acute leukemia. We find, for instance, that light has a dramatic effect on plants despite the absence of the four major photoreceptors, that bacterial-, fungal-, and viral-induced responses converge at later stages of attack, and that sample preparation procedures used in different hospitals have large effects on transcriptome patterns. We use canonical discriminant analysis to identify the genes associated with these patters and hierarchical clustering to find groups of coregulated genes that are easily visualized in a second round of correspondence analysis and ordered tables. The unconventional combination of standard descriptive multivariate methods offers a previously unrecognized tool to uncover unexpected information.


Dynamic phase extraction in high-SNR DAS based on UWFBGs without phase unwrapping using scalable homodyne demodulation in direct detection.
We demonstrate a Distributed Acoustic Sensor (DAS) based on Ultra-Weak Fiber Bragg Gratings (UWFBGs) using a scalable homodyne demodulation in direct detection. We show that a distributed interferometric system using delay and mixing of backscattering from consecutive identical gratings can be combined with a Phase-Generated Carrier Differentiate and Cross-Multiply (PGC-DCM) demodulation algorithm to perform dynamic measurements with high SNR, employing a simple narrowband laser and a pin photodiode. The proposed homodyne demodulation technique is suitable for real-time monitoring using distributed measurements, as it does not require computationally costly phase unwrapping common in conventional schemes and is robust against detrimental harmonic distortions, while not requiring additional mechanisms to handle division-by-zero operations. The demodulation scheme is also scalable, as it involves symmetric ordinary differentiation and integration operations suitable for processing with FPGA-based or analogue systems which, thanks to readily realizable schemes for implementing fractional order calculus, are also candidates for small-scale integration. We experimentally demonstrate the effectiveness of the technique by monitoring the dynamic response of a generic 2.5 kHz vibration applied to a PZT placed at the end of a sensing fiber comprised of a 1 km array of 200 UWFBGs each with a reflectivity of ~-43 dB written at a spacing of 5 m, with an SNR of ~34.52 dB.


The aging face.
Aging is an inevitable process. Most patients identify the concept of self-image in their facial appearance. The face is also our interface with the outside world. Changes in the face secondary to aging are the most apparent. Some aspects of aging are fairly uncontrollable, and these are largely based on hereditary factors. Other factors are somewhat controllable and are largely the result of exposure to the elements and harmful habits. The processes of facial aging are slowly becoming understood. Although different patients appear to age at different rates, biologic aging does not seem necessarily to follow chronologic aging in the same fashion in different individuals. However, a fundamental pattern as well as sequence in aging is fairly predictable and describable. The effects of ultraviolet radiation and involutional changes are manifested in the skin as wrinkles. Involutional changes in the suspensory structures in the face result in skin laxity and dependency. In this article, the various factors involved in aging are reviewed. The effects of these factors in different regions are discussed, with representative illustrations to represent concepts.


Modulation of energy and protein supplies in sequential feeding in laying hens.
Sequential feeding (SF) consists of splitting energy (E) and protein/calcium (P) fractions temporally, improving the feed conversion ratio (FCR) of hens compared with a continuous distribution during the day. In a previous study, the E fraction (with a low level of protein) was provided in the morning, whereas the P fraction (with low level of energy) was given in the afternoon. However, there is no clear evidence that a requirement in energy or proteins is connected to these distribution sequences, whereas the requirement for calcium is known to be required in the afternoon. To evaluate the effects on performances of the modulation of energy and protein supplies in SF, five different sequential treatments were offered: E0P0/E0P0; E+P+/E-P-; E+P-/E-P+; E0P+/E0P- and E+P0/E-P0 where E+ represents a high energy level, E0 a moderate one and E- a low one (with the same meaning for P regarding protein supply). Afternoon fractions were provided with particulate calcium. A total of 168 Hendrix hens were housed in individual cages from 20 to 39 weeks of age in two environmentally contrasted rooms. Feed intake in the morning and afternoon fractions, egg production, egg weight, BW and weight of digestive organs were recorded. No diet effect was observed concerning feed intake, egg production and BW. These results suggested that hens are not able to fit their feed intake on energy or protein level of fractions within half-day duration, whereas at the day scale same protein and energy intakes were observed. Moreover, the time of nutrient distribution in feeding did not seem to have an impact on birds' performances. These studies have also demonstrated that, despite strong environmental pressure, the hens with SF had attenuated performance but continue to produce eggs.


Insertion of ISPa1635 in ISCR1 Creates a Hybrid Promoter for blaPER-1 Resulting in Resistance to Novel β-lactam/β-lactamase Inhibitor Combinations and Cefiderocol.
Eleven blaPER-1-positive Pseudomonas aeruginosa clinical isolates showed variable susceptibility to ceftazidime-avibactam (CZA). The genetic contexts of blaPER-1 were identical (ISCR1-blaPER-1-gst) except for the ST697 isolate HS204 (ISCR1-ISPa1635-blaPER-1-gst). The insertion of ISPa1635 in ISCR1 upstream of blaPER-1 created a hybrid promoter, which elevated the blaPER-1 transcription level and resulted in increased resistance to CZA, ceftolozane-tazobactam, cefepime-zidebactam, and cefiderocol. Diversity in the promoter activity of blaPER-1 partially explains the variable susceptibility to CZA in PER-producing isolates.


Transcriptional activation and transformation by FosB protein require phosphorylation of the carboxyl-terminal activation domain.
The transcription factor AP-1, composed of Fos-Jun dimers, mediates some aspects of the cellular response to growth factors. Transcriptional activation and neoplastic transformation by FosB, a member of the Fos family of proteins, require the presence of a potent C-terminal activation domain. Here we show by mutational analysis that the FosB C-terminal domain has a proline-based motif that is essential for both of these functions. Phosphopeptide mapping experiments show that the C terminus of FosB is phosphorylated within a cluster of functionally redundant serine residues that is adjacent to this proline-based motif. Mutation of these serine residues to alanine severely reduces the ability of this region to function as an activation domain and inhibits the ability of FosB protein to function as a transforming protein. Several observations suggest that the kinase responsible for phosphorylation of these sites is distinct from the mitogen-activation protein kinases and stress-activated protein kinases. Our results show that transcriptional activation and neoplastic transformation by the FosB protein are dependent on phosphorylation within the C terminus. This form of control may provide a potential mechanism of signal integration at the level of a single transcription factor.


Automated identification of multiple seizure-related and interictal epileptiform event types in the EEG of mice.
Visual scoring of murine EEG signals is time-consuming and subject to low inter-observer reproducibility. The Racine scale for behavioral seizure severity does not provide information about interictal or sub-clinical epileptiform activity. An automated algorithm for murine EEG analysis was developed using total signal variation and wavelet decomposition to identify spike, seizure, and other abnormal signal types in single-channel EEG collected from kainic acid-treated mice. The algorithm was validated on multi-channel EEG collected from γ-butyrolacetone-treated mice experiencing absence seizures. The algorithm identified epileptiform activity with high fidelity compared to visual scoring, correctly classifying spikes and seizures with 99% accuracy and 91% precision. The algorithm correctly identifed a spike-wave discharge focus in an absence-type seizure recorded by 36 cortical electrodes. The algorithm provides a reliable and automated method for quantification of multiple classes of epileptiform activity within the murine EEG and is tunable to a variety of event types and seizure categories.


Transcriptome Analysis of Pseudomonas aeruginosa Cultured in Human Burn Wound Exudates.
Pseudomonas aeruginosa is a severe opportunistic pathogen and is one of the major causes of hard to treat burn wound infections. Herein we have used an RNA-seq transcriptomic approach to study the behavior of P. aeruginosa PAO1 growing directly on human burn wound exudate. A chemical analysis of compounds used by this bacterium, coupled with kinetics expression of central genes has allowed us to obtain a global view of P. aeruginosa physiological and metabolic changes occurring while growing on human burn wound exudate. In addition to the numerous virulence factors and their secretion systems, we have found that all iron acquisition mechanisms were overexpressed. Deletion and complementation with pyoverdine demonstrated that iron availability was a major limiting factor in burn wound exudate. The quorum sensing systems, known to be important for the virulence of P. aeruginosa, although moderately induced, were activated even at low cell density. Analysis of bacterial metabolism emphasized importance of lactate, lipid and collagen degradation pathways. Overall, this work allowed to designate, for the first time, a global view of P. aeruginosa characteristics while growing in human burn wound exudate and highlight the possible therapeutic approaches to combat P. aeruginosa burn wound infections.


Radiosensitivity of human colon cancer cell enhanced by immunoliposomal docetaxel.
To enhance the radiosensitivity of human colon cancer cells by docetaxel. Immunoliposomal docetaxel was prepared by coupling monoclonal antibody against carcinoembryonic antigen to cyanuric chloride at the PEG terminus of liposome. LoVo adenocarcinoma cell line was treated with immunoliposomal docetaxel or/and irradiation. MTT colorimetric assay was used to estimate cytotoxicity of immunoliposomal docetaxel and radiotoxicity. Cell cycle redistribution and apoptosis were determined with flow cytometry. Survivin expression in LoVo cells was verified by immunohistochemistry. D801 morphologic analysis system was used to semi-quantify immunohistochemical staining of survivin. Cytotoxicity was induced by immunoliposomal docetaxel alone in a dose-dependent manner. Immunoli-posomal docetaxel yielded a cytotoxicity effect at a low dose of 2 nmol/L. With a single dose irradiation, the relative surviving fraction of LoVo cells showed a dose-dependent response, but there were no significant changes as radiation delivered from 4 to 8 Gy. Compared with liposomal docetaxel or single dose irradiation, strongly radiopotentiating effects of immunoliposomal docetaxel on LoVo cells were observed. A low dose of immunoliposomal docetaxel could yield sufficient radiosensitivity. Immunoliposomal docetaxel were achieved both specificity of the conjugated antibody and drug radiosensitization. Combined with radiation, immunoliposomal docetaxel significantly increased the percentage of G(2)/M cells and induced apoptosis, but significantly decreased the percentage of cells in G(2)/G(1) and S phase by comparison with liposomal docetaxel. Immunohistochemical analysis showed that the brown stained survivin was mainly in cytoplasm of LoVo cells. Semi-quantitative analysis of the survivin immunostaining showed that the expression of survivin in LoVo cells under irradiation with immunoliposomal docetaxel was significantly decreased. Immunoliposomal docetaxel is strongly effective for target radiosensitation in LoVo colon carcinoma cells, and may offer the potential to improve local radiotherapy.


Physically effectiveness of beet pulp-based diets in dairy cows as assessed by responses of feed intake, digestibility, chewing activity and milk production.
Nine multiparous Holstein cows (DIM = 56 ± 5 day) were randomly assigned to three experimental diets containing fine-dried (T1), normal-dried (T2) and pelleted (T3) beet pulp (BP) at 12% on a dry matter basis in a 3 × 3 change over design in three 21 day periods to evaluate the effects of three types of BP. Bulk density, functional specific gravity and water-holding capacity of ration and intake were similar, but intake of physically effective fibre was different among treatments. Without significant differences, nutrients' digestibility in T1 trended to be higher than T2 and T3. Ruminal liquid pH and N-NH3 concentration were lower in T1 and higher in T3 than T2. Grinding and pelleting of BP increased and decreased volatile fatty acid concentration respectively. Grinding decreased acetate and propionate and increased butyrate and lactic acid. In contrary to pelleting, grinding of BP increased the particulate ruminal passage rate, but decreased ruminal mean retention time and lower compartment mean retention time. Pelleting of BP decreased ruminal passage rate, but increased ruminal mean retention time and lower compartment mean retention time. Grinding increased 5.64, 5.9 and 5.8% eating time, rumination and total chewing activity in comparison with normal BP respectively. Pelleting increased 2.7%, 16.3% and 10.0% eating time, rumination and total chewing activity in comparison with T2 respectively. Milk yield had no significant difference, but milk yield adjusted to 4% fat, and milk fat (kg/day), fat, protein, casein and total solid of milk (%) were affected by the treatments. Results from this experiment show that based on milk fat assay, grinding did not reduce effectiveness of BP, but pelleting significantly increased effectiveness of BP. In addition, based on total chewing activity, grinding did not reduce physically effectiveness of BP, but pelleting of BP increased its physically effectiveness.


[The excitotoxicity theory of glaucoma].
Glaucoma can be defined as a disease in which one of the pathophysiological consequences of raised intra-ocular pressure is damage of the optic nerve, and subsequently the loss of retinal ganglion cells (RGCs). One of the main aims of modern glaucoma therapy is to alter the intraocular pressure, either surgically or pharmacologically. Recently it was shown that the vitreous of glaucoma patients contains increased levels of glutamate (27 microM as compared to 11 microM in controls). This concentration of glutamate is sufficient to induce retinal ganglion cell death. The rise in intraocular pressure is probably the initial insult, which enhances the increase or release of glutamate. Although the increase in intravitreal glutamate levels is an accompanying feature of glaucoma, it could contribute to the loss of retinal ganglion cells in humans itself. Therefore, despite efficient control of intra-ocular pressure, RGC's loss will continue resulting in further visual impairment, if the toxic effect of glutamate is not blocked. If it would be possible to understand the mechanism leading to excessive vitreous levels of glutamate in glaucoma or to block its toxic effects, then the resulting visual loss could be retarded. This review discusses various proposed mechanisms leading to intraocular glutamate toxicity and the role of neuroprotection in this disease. (Literature search by Medline).


Update of the drug resistance mutations in HIV-1: Fall 2005.
The International AIDS Society-USA (IAS-USA) Drug Resistance Mutations Group is marking 5 years as an independent volunteer panel of experts focused on identifying key HIV-1 drug resistance mutations. The goal of the effort is to quickly deliver accurate and unbiased information on these mutations to HIV clinical practitioners. The October/November 2005 version of the IAS-USA Drug Resistance Mutations Figures replaces the version published in this journal in March/April 2005. The IAS-USA Drug Resistance Mutations Figures are designed for use in identifying mutations associated with viral resistance to antiretroviral drugs and in making therapeutic decisions. Care should be taken when using this list of mutations for surveillance or epidemiologic studies of transmission of drug-resistant virus. A number of amino acid substitutions, particularly minor mutations, represent polymorphisms that in isolation may not reflect prior drug selective pressure or reduced drug susceptibility.


Transmission of Influenza A in a Student Office Based on Realistic Person-to-Person Contact and Surface Touch Behaviour.
Influenza A viruses result in the deaths of hundreds of thousands of individuals worldwide each year. In this study, influenza A transmission in a graduate student office is simulated via long-range airborne, fomite, and close contact routes based on real data from more than 3500 person-to-person contacts and 127,000 surface touches obtained by video-camera. The long-range airborne, fomite and close contact routes contribute to 54.3%, 4.2% and 44.5% of influenza A infections, respectively. For the fomite route, 59.8%, 38.1% and 2.1% of viruses are transmitted to the hands of students from private surfaces around the infected students, the students themselves and other susceptible students, respectively. The intranasal dose via fomites of the students' bodies, belongings, computers, desks, chairs and public facilities are 8.0%, 6.8%, 13.2%, 57.8%, 9.3% and 4.9%, respectively. The intranasal dose does not monotonously increase or decrease with the virus transfer rate between hands and surfaces. Mask wearing is much more useful than hand washing for control of influenza A in the tested office setting. Regular cleaning of high-touch surfaces, which can reduce the infection risk by 2.14%, is recommended and is much more efficient than hand-washing.


The association between repetitive, self-injurious and aggressive behavior in children with severe intellectual disability.
We evaluated the independent association between adaptive behavior, communication and repetitive or ritualistic behaviors and self-injury, aggression and destructive behavior to identify potential early risk markers for challenging behaviors. Data were collected for 943 children (4-18 years, M = 10.88) with severe intellectual disabilities. Odds ratio analyses revealed that these characteristics generated risk indices ranging from 2 to 31 for the presence and severity of challenging behaviors. Logistic regressions revealed that high frequency repetitive or ritualistic behavior was associated with a 16 times greater risk of severe self-injury and a 12 times greater risk of showing two or more severe challenging behaviors. High frequency repetitive or ritualistic behaviors independently predict challenging behavior and have the potential to be early risk markers for self-injury and aggression of clinical significance.


Physical and genetic characterization of a 75-kilobase deletion associated with al, a recessive lethal allele at the mouse agouti locus.
The agouti locus (A) of the mouse determines the timing and type of pigment deposition in the growing hair bulb, and several alleles at this locus are lethal when homozygous. Apparent instances of intragenic recombination and complementation between different recessive lethal alleles have suggested that the locus has a complex structure. We have begun to investigate the molecular basis of agouti gene action and recessive lethality by using a series of genetically linked DNA probes and pulsed field gel electrophoresis to detect structural alterations in radiation-induced agouti mutations. Hybridization probes from the Src and Emv-15 loci do not reveal molecular alterations in DNA corresponding to the ae, ax, and al alleles, but a probe from the parotid secretory protein gene (Psp) detects a 75-kilobase (kb) deletion in DNA containing the non-agouti lethal allele (al). The deletion is defined by a 75-kb reduction in the size of BssHII, NotI, NruI and SacII high molecular weight restriction fragments detected with the Psp probe and is located between 25 kb and 575 kb from Psp coding sequences. Because the genetic distance between A and Emv-15 is much less than A and Psp, there may be a preferred site of recombination close to Psp, or suppression of recombination between A and Emv-15. The al deletion has allowed us to determine the genotype of mice heterozygous for different recessive lethal alleles. We find that three different recessive lethal complementation groups are present at the agouti locus, two of which are contained within the al deletion.


Pathways for self-tolerance and the treatment of autoimmune diseases.
Antigen delivers both immunogenic and tolerogenic signals to lymphocytes. The outcome of antigen exposure represents a complex integration of the timing of antigen binding with signals from many other immunogenic and tolerogenic costimulatory pathways. A road map of these signalling pathways is only beginning to be charted, revealing the mechansim of action and limitations of current immunotherapeutic agents and the points of attack for new agents. Ciclosporin and tacrolimus interfere with tolerogenic signals from antigen in addition to blocking immunogenic signals, thus preventing active establishment of tolerance. Corticosteroids inhibit a key immunogenic pathway, NFkappaB, and more specific inhibitors of this pathway may allow tolerance to be actively established while immune responses are blocked. New experimental therapies aim to mimic tolerogenic antigen signals by chronically stimulating antigen receptors with antigen or antibodies to the receptor, or aim to block costimulatory pathways involving CD40 ligand, B7, or interleukin 2. Obtaining the desired response with these strategies is unpredictable because many of these signals have both tolerogenic and immunogenic roles. The cause of autoimune diseases has been determined for several rare monogenic disorders, revealing inherited deficiencies in tolerogenic costimulatory pathways such as FAS. Common autoimmune disorders may have a biochemically related pathogenesis.


Chronic AMP-kinase activation with AICAR reduces adiposity by remodeling adipocyte metabolism and increasing leptin sensitivity.
This study investigated the effect of chronic AMP-kinase (AMPK) activation with 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) on white adipose tissue (WAT) metabolism and the implications for visceral (VC) and subcutaneous (SC) adiposity, whole body-energy homeostasis, and hypothalamic leptin sensitivity. Male Wistar rats received daily single intraperitoneal injections of either saline or AICAR (0.7g/kg body weight) for 4 and 8 weeks and were pair-fed throughout the study. AICAR-treated rats had reduced adiposity with increased mitochondrial density in VC and SC fat pads, which was accompanied by reduced circulating leptin and time-dependent and depot-specific regulation of AMPK phosphorylation and FA oxidation. Interestingly, the anorectic effect to exogenous leptin was more pronounced in AICAR-treated animals than controls. This corresponded to reductions in hypothalamic AMPK phosphorylation and suppressor of cytokine signaling 3 content, whereas signal transducer and activator of transcription 3 phosphorylation was either unchanged or increased at 4 and 8 weeks in AICAR-treated rats. Ambulatory activity and whole-body energy expenditure (EE) were also increased with AICAR treatment. Altogether, chronic AICAR-induced AMPK activation increased WAT oxidative machinery, whole-body EE, and hypothalamic leptin sensitivity. This led to significant reductions in VC and SC adiposity without inducing energy-sparing mechanisms that oppose long-term fat loss.


Comparative analyses of six solanaceous transcriptomes reveal a high degree of sequence conservation and species-specific transcripts.
The Solanaceae is a family of closely related species with diverse phenotypes that have been exploited for agronomic purposes. Previous studies involving a small number of genes suggested sequence conservation across the Solanaceae. The availability of large collections of Expressed Sequence Tags (ESTs) for the Solanaceae now provides the opportunity to assess sequence conservation and divergence on a genomic scale. All available ESTs and Expressed Transcripts (ETs), 449,224 sequences for six Solanaceae species (potato, tomato, pepper, petunia, tobacco and Nicotiana benthamiana), were clustered and assembled into gene indices. Examination of gene ontologies revealed that the transcripts within the gene indices encode a similar suite of biological processes. Although the ESTs and ETs were derived from a variety of tissues, 55-81% of the sequences had significant similarity at the nucleotide level with sequences among the six species. Putative orthologs could be identified for 28-58% of the sequences. This high degree of sequence conservation was supported by expression profiling using heterologous hybridizations to potato cDNA arrays that showed similar expression patterns in mature leaves for all six solanaceous species. 16-19% of the transcripts within the six Solanaceae gene indices did not have matches among Solanaceae, Arabidopsis, rice or 21 other plant gene indices. Results from this genome scale analysis confirmed a high level of sequence conservation at the nucleotide level of the coding sequence among Solanaceae. Additionally, the results indicated that part of the Solanaceae transcriptome is likely to be unique for each species.


Circulatory functions during immersion and breath-hold dives in humans.
An unusual circulatory state exists in humans immersed in water at or near 35 degrees C. This circulatory state is characterized by a persistent and elevated stroke index (SI) but heart rate (HR) changes little; hence an increased cardiac index (CI) results. In comparison of measurements in air and immersion in water up to the neck level that are based on the weighted averages of 45 subjects from 8 studies, SI increased by 29%, CI increased by 24%, and HR decreased by 6%. Evidence is presented to support the finding that the increase in stroke volume is a result of enhanced preload of the heart, and that alterations in afterload and contractility play an unimportant role in affecting cardiac performance during immersion in thermoneutral water up to the level of the neck. The circulatory state represents hyperperfusion, since there is no concurrent rise in metabolic demand or heat stress. To what extent this preexisting circulatory state affects the cardiovascular responses to breath-hold (BH) diving is in most part unknown. However, the BH bradycardiac responses are more potent in a natural setting than in the laboratory where the breath hold is performed with only face immersion in water. In contrast, in the natural setting, the divers perform BH dives while they are immersed up to the neck in cold water (much lower than 35 degrees C) and are exercising. It is desirable in future studies to compare systematically other aspects of circulatory responses to BH dives while immersed in water and while exercising.


[Monoclonal gammopathy of undetermined significance : when and why to look for them ?]
Monoclonal gammopathy of undetermined significance (MGUS) is an asymptomatic premalignant plasma cell disorder. MGUS is a common disorder and the diagnosis is often made accidentally when a protein electrophoresis is performed in a routine blood test or during a biological assessment for other conditions. In the absence of biological abnormalities or clinical symptoms suggesting a plasma or lymphoplasma-cell disorder, there is no indication for routine screening of the monoclonal protein. When MGUS is diagnosed, the risk of transformation into myeloma or other lymphoproliferative disorders is estimated at 1 % per year. MGUS can also be associated with diseases that are not malignant disorders and in some cases, the monoclonal gammopathy is the witness of another rare but severe disorder wich will be critical not to be missdiagnosed.


A new process for preparation of soybean protein concentrate with hexane-aqueous ethanol mixed solvents.
A new process for the preparation of soybean protein concentrate (SPC) by directly extracting full-fat soy flour with a mixture of hexane and aqueous ethanol was established. Compared with conventional methods, it has some advantages, such as saving energy and reducing protein denaturation caused by heat action during solvent recovery, because this process saves one step of solvent recovery. The effects of aqueous ethanol concentration and the mixure ratio (hexane to ethanol) on the degree of protein denaturation and product quality were investigated, on the basis of which the orthogonal tests were performed. The optimum technical parameters were obtained by analyzing the results of the orthogonal tests with statistical methods. We found that SPC can be obtained by extracting full-fat soy flour under the following conditions: mixture ratio hexane: 90% ethanol, 9:1, v/v; extraction temperature, 45 degrees C; ratio of solid to solvents, (1:2 w/v); and 5 repeated extractions (15 min each time). The results of quality analysis showed that solubility of the product was improved significantly [nitrogen solubility index (NSI) 46.6%] compared with that for ethanol washing of protein concentrate (NSI 8.7%).


Interference of maltose, icodextrin, galactose, or xylose with some blood glucose monitoring systems.
Maltose, a disaccharide composed of two glucose molecules, is used in a number of biological preparations as a stabilizing agent or osmolality regulator. Icodextrin, which is converted to maltose, is present in a peritoneal dialysis solution. Galactose and xylose are found in some foods, herbs, and dietary supplements; they are also used in diagnostic tests. When some blood glucose monitoring systems are used--specifically, those that use test strips containing the enzymes glucose dehydrogenase-pyrroloquinolinequinone or glucose dye oxidoreductase--in patients receiving maltose, icodextrin, galactose, or xylose, interference of blood glucose levels can occur. Maltose, icodextrin, galactose, and xylose are misinterpreted as glucose, which can result in erroneously elevated serum glucose levels. This interference can result in the administration of insulin, which may lead to hypoglycemia. In severe cases of hypoglycemia, deaths have occurred. If patients are receiving maltose, icodextrin, galactose, or xylose, clinicians must review the package inserts of all test strips to determine the type of glucose monitoring system being used and to use only those systems whose tests strips contain glucose oxidase, glucose dehydrogenase-nicotinamide adenine dinucleotide, or glucose dehydrogenase-flavin adenine dinucleotide.


Stress induced birefringence in hybrid TIR/PBG guiding solid photonic crystal fibers.
We report on two types of polarization maintaining solid photonic crystal fibers that guide light by a combination of a photonic bandgap and total internal reflection. Group and phase birefringence are studied experimentally and numerically for stress-applying parts made from B-doped and F-doped silica. The stress field originating from Ge-doped cladding rods is shown to interfere with the stress field from the B-doped and F-doped rods. Since the differential expansion coefficients of B-doped and F-doped silica have opposite signs this interference is either destructive or constructive. Consequently, we found that the fiber with F-doped stress applying parts has the highest modal phase birefringence, and polarization cross talk is characterized by an h-parameter below 310(-5) m(-1).


Mitochondrial-derived reactive oxygen species (ROS) play a causal role in aging-related intervertebral disc degeneration.
Oxidative damage is a well-established driver of aging. Evidence of oxidative stress exists in aged and degenerated discs, but it is unclear how it affects disc metabolism. In this study, we first determined whether oxidative stress negatively impacts disc matrix metabolism using disc organotypic and cell cultures. Mouse disc organotypic culture grown at atmospheric oxygen (20% O(2)) exhibited perturbed disc matrix homeostasis, including reduced proteoglycan synthesis and enhanced expression of matrix metalloproteinases, compared to discs grown at low oxygen levels (5% O(2)). Human disc cells grown at 20% O(2) showed increased levels of mitochondrial-derived superoxide anions and perturbed matrix homeostasis. Treatment of disc cells with the mitochondria-targeted reactive oxygen species (ROS) scavenger XJB-5-131 blunted the adverse effects caused by 20% O(2). Importantly, we demonstrated that treatment of accelerated aging Ercc1(-/Δ) mice, previously established to be a useful in vivo model to study age-related intervertebral disc degeneration (IDD), also resulted in improved disc total glycosaminoglycan content and proteoglycan synthesis. This demonstrates that mitochondrial-derived ROS contributes to age-associated IDD in Ercc1(-/Δ) mice. Collectively, these data provide strong experimental evidence that mitochondrial-derived ROS play a causal role in driving changes linked to aging-related IDD and a potentially important role for radical scavengers in preventing IDD.


Role of TEM-1 β-Lactamase in the Predominance of Ampicillin-Sulbactam-Nonsusceptible Escherichia coli in Japan.
We investigated the epidemiology and resistance mechanisms of ampicillin-sulbactam-nonsusceptible Escherichia coli, focusing on the role of the TEM-1 β-lactamase. We collected all nonduplicate E. coli clinical isolates at 10 Japanese hospitals during December 2014 and examined their antimicrobial susceptibility, β-lactamases, TEM-1 transferability, TEM-1 β-lactamase activity, outer membrane protein profile, membrane permeability, and clonal genotypes. Among the 329 isolates collected, 95 were ampicillin-sulbactam nonsusceptible. Of these ampicillin-sulbactam-nonsusceptible isolates, β-lactamases conferring resistance to sulbactam, such as AmpC, were present in 33%. Hyperproduction of sulbactam-susceptible β-lactamases, TEMs with a strong promoter, were rare (5%). The remaining 59 isolates (62%) had only sulbactam-susceptible β-lactamases, including TEM-1 with a wild-type promoter (n = 28), CTX-Ms (n = 13), or both (n = 17). All 45 transconjugants from 96 donors with TEM-1 had higher ampicillin-sulbactam MICs (4 to 96 mg/liter) than the recipient (2 mg/liter). In donors with only TEM-1, TEM-1 activity correlated with the 50% inhibitory concentration of sulbactam and ampicillin-sulbactam MICs. The decreased membrane permeation of sulbactam was associated with an increased ampicillin-sulbactam MIC. The reduced permeation was partly attributable to deficient outer membrane proteins, which were observed in 57% of the ampicillin-sulbactam-nonsusceptible isolates with only TEM-1 and a wild-type promoter. Sequence type 131 (ST131) was the most common clonal type (52%). TEM-1 with a wild-type promoter primarily contributed to ampicillin-sulbactam nonsusceptibility in E. coli, with the partial support of other mechanisms, such as reduced permeation. Conjugative TEM-1 and the clonal spread of ST131 may contribute to the prevalence of Japanese ampicillin-sulbactam-nonsusceptible isolates.


Thermally activated switching in the presence of non-Gaussian noise.
We study the effect of a non-Gaussian noise on interstate switching activated primarily by Gaussian noise. Even weak non-Gaussian noise can strongly change the switching rate. The effect is determined by all moments of the noise distribution. It is expressed in a closed form in terms of the noise characteristic functional. The analytical results are compared with the results of simulations for an overdamped system driven by white Gaussian noise and a Poisson noise. Switching induced by a purely Poisson noise is also discussed.


Fertility and ageing.
The late 20th century trend to delay birth of the first child until the age at which female fecundity or reproductive capacity is lower has increased the incidence of age-related infertility. The trend and its consequences have also stimulated interest in the possible factors in the female and the male that may contribute to the decline in fecundity with age; in the means that exist to predict fecundity; and in the consequences for pregnancy and childbirth. In the female, the number of oocytes decreases with age until the menopause. Oocyte quality also diminishes, due in part to increased aneuploidy because of factors such as changes in spindle integrity. Although older male age affects the likelihood of conception, abnormalities in sperm chromosomes and in some components of the semen analysis are less important than the frequency of intercourse. Age is as accurate as any other predictor of conception with assisted reproductive technology. The decline in fecundity becomes clinically relevant when women reach their mid-30s, when even assisted reproduction treatment cannot compensate for the decline in fecundity associated with delaying attempts at conceiving. Pregnancies among women aged >40 years are associated with more non-severe complications, more premature births, more congenital malformations and more interventions at birth.


Most m6A RNA Modifications in Protein-Coding Regions Are Evolutionarily Unconserved and Likely Nonfunctional.
Methylation of the adenosine base at the nitrogen-6 position (m6A) is the most prevalent internal posttranscriptional modification of mRNAs in many eukaryotes. Despite the rapid progress in the transcriptome-wide mapping of m6As, identification of proteins responsible for writing, reading, and erasing m6As, and elucidation of m6A functions in splicing, RNA stability, translation, and other processes, it is unknown whether most observed m6A modifications are functional. To address this question, we respectively analyze the evolutionary conservation of yeast and human m6As in protein-coding regions. Relative to comparable unmethylated As, m6As are overall no more conserved in yeasts and only slightly more conserved in mammals. Furthermore, yeast m6As and comparable unmethylated As have no significant difference in single nucleotide polymorphism (SNP) density or SNP site frequency spectrum. The same is true in human. The methylation status of a gene, not necessarily the specific sites methylated in the gene, is subject to purifying selection for no more than ∼20% of m6A-modified genes. These observations suggest that most m6A modifications in protein-coding regions are nonfunctional and nonadaptive, probably resulting from off-target activities of m6A methyltransferases. In addition, our reanalysis invalidates the recent claim of positive selection for newly acquired m6A modifications in human evolution. Regarding the small number of evolutionarily conserved m6As, evidence suggests that a large proportion of them are likely functional; they should be prioritized in future functional characterizations of m6As. Together, these findings have important implications for understanding the biological significance of m6A and other posttranscriptional modifications.


[Mechanism of membrane fouling and filtration characteristics in a membrane bioreactor for industrial wastewater treatment].
The influence of mixed liquor suspended solids (MLSS), soluble microbial product (SMP), extracellular polymeric substance (EPS), colloidal particles and other factors contributed to membrane fouling was analyzed in this pilot test by membrane bioreactor (MBR) process for the leather printing and dyeing industrial park mixed wastewater treatment. The results showed that slight membrane fouling occurred after 120-day experiment with an observable increase in membrane resistance R20 from 1.5 x 10(12) m(-1) to 1.8 x 10(12) m(-1). Also, a linear correlation was found between the proportion of colloidal particles concentration in TOC of MBR former solution and membrane filtration resistance change. However, the change of MLSS, SMP, EPS and other factors was not correlated with the membrane filtration resistance change. Therefore, the colloidal particle was considered to be the main factor causing membrane fouling, which attached to the membrane surface and deposited to block the membrane pore.


Effect of rotavirus vaccine on reducing acute gastroenteritis in a large outpatient pediatric network.
To measure the effect of rotavirus vaccine (RVV) on acute gastroenteritis (AGE) managed by primary care physicians in the first 2 rotatvirus seasons following the introduction of RVV. Retrospective cohort study. Practice-based network composed of 33 ambulatory pediatric practices in Philadelphia, Pennsylvania. All children born from February 22, 2006 (date of RVV licensure), through February 29, 2008, and who received care at any network site. Receipt of RVV. The primary outcomes were AGE-related office visits, telephone calls, and episodes (composite outcome consisting of all visits and calls within a 10-day period). Rates of AGE visits in the 2 rotavirus seasons following the introduction of RVV steadily decreased from 3.0 AGE visits per 100 total office visits in the 2005 season to 1.8 in the 2008 season. In 2007, vaccinations were administered to 9351 of 13 951 vaccine-eligible children (67.0%), and in 2008, they were administered to 9958 of 10 728 (92.8%). Among RVV-immunized children in 2007, AGE calls and episodes were significantly reduced with vaccine effectiveness of 53% and 46%, respectively. No significant difference was seen between RVV-immunized and RVV-nonimmunized children for any outcome in 2008. Rotavirus vaccine was associated with a significant reduction in outpatient AGE calls and episodes among immunized children in our network in 2007. Despite a reduction in winter AGE rates in the network, no difference was detected between RVV-immunized and RVV-nonimmunized children for any outcome in 2008. Further study is needed to understand the lack of vaccine effect in 2008.


Pain Management in Acute Pancreatitis: A Systematic Review and Meta-Analysis of Randomised Controlled Trials.
Background: Pain management is an important priority in the treatment of acute pancreatitis (AP). Current evidence and guideline recommendations are inconsistent on the most effective analgesic protocol. This systematic review and meta-analysis of randomised controlled trials (RCTs) aimed to compare the safety and efficacy of analgesics for pain relief in AP. Methods: A literature search was performed to identify all RCTs assessing analgesics in patients with AP. The primary outcome was the number of participants who needed rescue analgesia. Study quality was assessed using Jadad score. Pooled odds ratios (ORs) or weighted mean differences (WMDs) with 95% confidence intervals (CI) were analysed using a random-effects model. Results: Twelve studies comprising 699 patients with AP (83% mild AP) were analysed. The tested analgesics significantly decreased the need for rescue analgesia (3 studies, OR.36, 95% CI 0.21 to 0.60) vs. placebo or conventional treatment. The analgesics also improved the pain score [Visual Analogue Scale (Δ-VAS)] at 24 h (WMD 18.46, 0.84 to 36.07) and by the 3rd to 7th days (WMD 11.57, 0.87 to 22.28). Opioids vs. non-opioids were associated with a decrease in the need for rescue analgesia (6 studies, OR 0.25, 95% CI 0.07 to 0.86, p = 0.03) but without significance in pain score. In subgroup analyses, opioids were similar to non-steroidal anti-inflammatory drugs (NSAIDs) regarding the primary outcome (4 studies, OR 0.56, 95% CI 0.24 to 1.32, p = 0.18). There were no significant differences in other clinical outcomes and rate of adverse events. Other studies, comparing epidural anaesthesia vs. patient-controlled analgesia and opioid (buprenorphine) vs. opioid (pethidine) did not show significant difference in primary outcome. Study quality issues significantly contributed to overall study heterogeneity. Conclusions: NSAIDs and opioids are equally effective in decreasing the need for rescue analgesia in patients with mild AP. The relative paucity of trials and high-quality data in this setting is notable and the optimal analgesic strategy for patients with moderately severe and severe AP still requires to be determined.


Asparagine-linked glycoprotein biosynthesis in rat epididymis. Presence of a mannosidase II-like enzyme.
Previous studies from this laboratory using p-nitrophenyl alpha-D-mannoside (p-NPM) as substrate provided no evidence for the presence of mannosidase II in the rat epididymis [Skudlarek & Orgebin-Crist (1988) J. Reprod. Fertil. 84, 611-617]. However, rat epididymal epithelial cells cultured in the presence of swainsonine, an inhibitor of mannosidase II, produce abnormally processed N-linked glycoproteins containing hybrid-type oligosaccharides instead of complex-type [Tulsiani, Skudlarek & Orgebin-Crist (1990) Biol. Reprod. 43, 130-138], a result providing indirect evidence for the presence of mannosidase II-like enzyme in rat epididymis. In the studies described here, we present evidence for the occurrence of this processing enzyme in rat epididymal Golgi membranes. This enzyme is an integral Golgi membrane component. Like liver mannosidase II, the epididymal enzyme cleaves alpha 1,3- and alpha 1,6-linked mannosyl residues from GlcNAcMan5GlcNAc. However, unlike liver mannosidase II, the epididymal enzyme shows no activity towards the synthetic substrate, p-NPM. The epididymal mannosidase cross-reacts with liver anti-(mannosidase II) antibody, a result suggesting that the two enzymes share a common antigenic site(s). Immunoblotting studies following resolution of liver and epididymal Golgi membranes on SDS/PAGE show that, whereas the liver mannosidase II was resolved as a doublet of Mr 120,000 and 122,000, only the Mr 120,000 band was observed in the epididymal Golgi membranes. Immunoblotting of the Golgi-rich fractions, resolved under non-denaturing conditions, showed different patterns of charge and/or size isomers from the two tissues. These studies demonstrate tissue-specific differences in processing enzymes with similar function.


Migraine: diagnosis, management, and new treatment options.
The safety and tolerability of medications used to treat acute migraine attacks are summarized, the classification of headaches and the causes of and diagnostic criteria for migraine are reviewed, and the clinical tolerability profiles and therapeutic benefits of second-generation triptans are presented. Migraine is a paroxysmal disorder characterized by attacks of headache, nausea, vomiting, photophobia, and phonophobia. Drugs used to prevent migraine and those that effectively treat acute migraine attacks are readily available. Mild or moderate migraines are often treated with aspirin, acetaminophen, nonsteroidal anti-inflammatory drugs, antiemetic drugs, or isometheptene. Triptans (5-HT1 receptor agonists) are used to treat moderate or severe migraine and when nonspecific medications have been ineffective. Because sumatriptan, the first triptan used, is effective but can induce adverse events, second-generation triptans (zolmitriptan, naratriptan, rizatriptan, and almotriptan) were developed to increase the benefit-to-risk ratio in migraine management. Important pharmacologic, pharmacokinetic, and clinical differences exist among those drugs, but the tolerability profile of the newer triptans is very good, and they provide rapid relief from headache and sustained duration of effect. Primary care physicians must manage migraine patients with treatments that demonstrate a balance between efficacy and tolerability.


[Diagnostics of the skeletal massiveness and human somatotype using hand bones].
The skeleton massiveness (SM) and the somatotype of human constitution were determined on the basis of osteometry of bones of carpal and metacarpal bones (MB) and of phalanxes. Seventy male and 13 female skeletons from the collection of the chair for anthropology, Moscow State University, were investigated. Described are the results of examinations of 8 carpal bones made according to 3 signs (length, width, and height), and of 5 metacarpal bones made according to 4 signs (length, base and head width, and base height); investigation findings of finger phalanxes (in full) are also presented. Methods of current multidimensional statistics were used within the case study, i.e. related with the key components--for SM specification and the discriminative analysis--for constitution specification. The SM determination accuracy according to type A was 40%, according to type B--80%, according to type C--37.5% and according to type D--52.9%. The classification accuracy of constitutions by carpal bones was 50.0%, by MB--46.4%, and by MB plus finger phalanxes--48.1%. It is pointed out that it was for the first time that the elaborated quantitative criteria of osteometry of hand bones could be used in the expertise practice for the purpose of personality identification by osseous remains.


Geospatial Investigation of Nigerian Honey and Detection of Anti-Enteric Biomarker.
Geospatial mapping and antibacterial biomarkers were investigated in Nigerian honey used for therapeutic purposes in several communities affected with prevalent antibiotic-resistant enteric bacilli. Randomly collected enteric bacilli from faecal samples were biotyped and phenotypically assayed for antibiotic resistance and profiled for R plasmids. R plasmid molecular weight and multiantibiotic resistance index (MARI) relatedness were evaluated for resistance among phylogroups. Honey cidal activity, time kill kinetics, and bioactive markers were determined and analysed for geospatial distribution. More than 30% enteric biotypes were resistant to cotrimoxazole, ciprofloxacin, and tetracycline at MIC ≥16 μg/ml (P=0.004). Two unrelated cluster complexes with diverse antibiotic resistance indices expressed high molecular weight plasmid (14.17 kbp) with 0.73 MARI to two classes of antibiotics. Among the resistant bacilli, only 24.3% (MIC90 500 mg/mL) and 8.1% (MBC90 1000 mg/mL) were susceptible to honey with evidence of 14.85% and 5.94% significant viable reduction at 2 × MIC to less than 2.50 Log10 CFU/mL (P < 0.05). Only alkaloids significantly regressed (P=0.028) with susceptibility of resistant bacilli significantly correlate with bacteria inhibition (r = 0.534, P=0.049) at optimal cutoff limit of 0.32 mg/ml. Antibacterial honey with significant alkaloid biomarkers was detected at 3°10'0-3°30'0E and 6°30'0-7°30'0N of Southwest Nigeria. Spatial mapping evidently indicated variation in honey physicochemical and bioactive compounds and identified geographical locations suitable for production of anti-enteric honey rich in alkaloids marker required for prevention and treatment of resistant enteric bacilli infections.


Beyond intuitive modeling: combining biophysical models with innovative experiments to move the circadian clock field forward.
Two major approaches have been used to model circadian clocks. Qualitative modeling, used prior to the recent wealth of detailed molecular knowledge, makes general predictions but cannot provide detailed mechanistic insights. The more recent biophysical approach, on the other hand, incorporates the biochemical events that drive the clock and can make detailed and testable molecular predictions. These predictions are being tested using new experimental techniques that measure reaction kinetics and the behavior of individual cells. A joint modeling and experimental approach has recently been used to understand how mutations affecting phosphorylation can lead to a short circadian period in tau mutant hamsters and in humans with familial advanced sleep phase syndrome (FASPS). Another recent study has revealed novel single-cell phenotypes of clock gene mutations, demanding revision of current biophysical models yet validating certain model predictions that were previously overlooked. A new paradigm for clock research is emerging in which modeling inspires new experimental efforts, experimental data inspire new modeling efforts, and joint modeling/experimental studies lead to a deeper understanding of mammalian circadian rhythms.


Crystal structure of the ternary complex of TCR, MHC class I and lipopeptides.
The covalent conjugation of a 14-carbon fatty acid (myristic acid) to the N-terminal Gly residue, termed N-myristoylation, occurs in some viral proteins to dictate their pathological function. This protein lipidation reaction, however, is monitored by host cytotoxic T lymphocytes that are capable of recognizing N-terminal lipopeptide fragments in the context of major histocompatibility complex (MHC) class I molecules. In a rhesus model of human AIDS, for example, the classical MHC class I allomorph, Mamu-B*05104, was shown to bind SIV Nef-derived 4-mer lipopeptides (myristic acid-Gly-Gly-Ala-Ile; C14nef4) and present them to the CD8+ T-cell line, SN45. These lipopeptides accommodated in MHC class I molecules expose much shorter peptide chains than conventional MHC class I-presented 8-10-mer peptides, and the molecular mechanisms by which αβ T-cell receptors (TCRs) recognize lipopeptides currently remain unclear. An X-ray crystallographic analysis of the SN45 TCR α and β heterodimer in a form that was co-crystallized with the C14nef4-bound Mamu-B*05104 complex indicated that the amide group of the N-myristoylated glycine residue offered a primary T-cell epitope by establishing a sole hydrogen bond between its nitrogen atom and the side chain of Glu at position 101 of CDR3β. Accordingly, the Glu to Ala mutation at this position resulted in the loss of lipopeptide recognition. On the other hand, TCRs were positioned remotely from the peptide portion of C14nef4, and strong interactions were not observed. Thus, these observations provide novel structural insights into lipopeptide recognition by TCRs, which contrast sharply with the general molecular principle of peptide recognition.


Queensland Centre for Mental Health Research: the first 17 years.
To reflect on the establishment and evolution of the Queensland Centre for Mental Health Research. Narrative historical review. First established as an inpatient research unit in December 1987, the focus of the Centre evolved in concert with the skills of the staff. After the structure was revised in 1996 and 1999, the Centre has evolved into a group with four main research streams--epidemiology, developmental neurobiology, genetics and policy and economics. Although the group maintains a strong focus on serious mental disorders such as schizophrenia, our policy and economic work has a wider perspective. The Queensland Centre for Mental Health Research is based in an historic mental health service, with laboratories in collaborating universities and institutes. Key lessons learnt by the group along the way relate to the importance of focusing on a restricted range of research topics in order to build a critical mass. Given a facilitating environment, hospital-based research groups can prosper. Over the last 17 years, a cost-efficient, focused and productive research group has evolved that has made contributions to international research.


Bacterial cytokinesis: From Z ring to divisome.
Ancestral homologues of the major eukaryotic cytoskeletal families, tubulin and actin, play critical roles in cytokinesis of bacterial cells. FtsZ is the ancestral homologue of tubulin and assembles into the Z ring that determines the division plane. FtsA, a member of the actin family, is involved in coordinating cell wall synthesis during cytokinesis. FtsA assists in the formation of the Z ring and also has a critical role in recruiting downstream division proteins to the Z ring to generate the divisome that divides the cell. Spatial regulation of cytokinesis occurs at the stage of Z ring assembly and regulation of cell size occurs at this stage or during Z ring maturation.


Corrections and clarifications.
In the table in the article "Can sustainable farming win the battle of the bottom line?" by Bob Holmes (Environment & the Economy, 25 June, p. 1893), the second and third lines in the legend should have read, "Alt. #1: conventional corn-soybean rotation with reduced tillage. Alt. #2: organic corn-beans-(wheat + clover)-clover-corn rotation with reduced tillage."


No initial therapy for stage III and IV non-Hodgkin's lymphomas of favorable histologic types.
The question of whether initial treatment is necessary in relatively asymptomatic patients with stage III and IV non-Hodgkin's lymphomas of favorable histologic types was studied by retrospective analysis. Two groups of patients were studied: [1] 44 nonprotocol patients, followed since 1963, in whom initial treatment was withheld until required to evaluate the pace of disease and the necessity of treatment; and [2] 112 previously untreated patients who have participated in prospectively randomized clinical trials since 1971. For all 44 "deferred" patients, the median time before requiring treatment was 31 months, and there have been 19 patients who have not yet required therapy for periods of 3 to 104 months. The median actuarial survival for all 44 patients was 121 months. At 4 years, the actuarial survival of the 44 patients with deferred treatment is 77.3%, compared with 83.2% for the 112 protocol patients (P = 0.60). Careful observation without initiation of therapy is an appropriate option in the management of patients with relatively asymptomatic advanced non-Hodgkin's lymphomas of favorable histologic types.


Integrated Proteomic and Phosphoproteomic Analysis of the Hippocampus in a Mouse Model of Early Life Inflammation.
Inflammation in early life is a risk factor for the development of neuropsychiatric diseases later in adolescence and adulthood, yet the underlying mechanism remains elusive. In the present study, we performed an integrated proteomic and phosphoproteomic analysis of the hippocampus to identify potential molecular mechanisms of early life inflammation-induced cognitive impairment. Both female and male mice received a single intraperitoneal injection of 100 μg/kg lipopolysaccharide (LPS) on postnatal day 10 (P10). Behavioral tests, including open field, elevated plus-maze, and Y-maze tests, were performed on P39, P40, and P41, respectively. After behavioral tests, male mice were sacrificed. The whole brain tissues and the hippocampi were harvested on P42 for proteomic, phosphoproteomic, Western blot, and Golgi staining. Early life LPS exposure induced cognitive impairment in male mice but not in female mice, as assessed by the Y-maze test. Therefore, following biochemical tests were conducted on male mice. By proteomic analysis, 13 proteins in LPS group exhibited differential expression. Among these, 9 proteins were upregulated and 4 proteins were downregulated. For phosphoproteomic analysis, a total of 518 phosphopeptides were identified, of which 316 phosphopeptides were upregulated and 202 phosphopeptides were downregulated in the LPS group compared with the control group. Furthermore, KEGG analysis indicated that early life LPS exposure affected the glutamatergic synapse and neuroactive ligand-receptor interaction, which were associated with synaptic function and energy metabolism. Increased level of brain protein i3 (Bri3), decreased levels of PSD-95 and mGLUR5, and dendritic spine loss after early life LPS exposure further confirmed the findings of proteomic and phosphoproteomic analysis. Our findings demonstrated that neuroinflammation and impaired synapse may be involved in early life inflammation-induced cognitive impairment. Future studies are required to confirm our preliminary results.


Reductive elimination pathway for homocysteine to methionine conversion in cobalamin-dependent methionine synthase.
Density functional theory has been applied to investigate the methyl transfer from methylcobalamin (MeCbl) cofactor to homocysteine (Hcy) as catalyzed by methionine synthase (MetH). Specifically, the S(N)2 and the reductive elimination pathways have been probed as the possible mechanistic pathways for the methyl transfer reaction. The calculations indicate that the activation barrier for the reductive elimination reaction (24.4 kcal mol(-1)) is almost four times higher than that for the S(N)2 reaction (7.3 kcal mol(-1)). This high energy demand of the reductive elimination pathway is rooted in the structural distortion of the corrin ring that is induced en route to the formation of the triangular transition state. Furthermore, the reductive elimination reaction demands the syn accommodation of the methyl group and the substrate over the upper face of the corrin ring, which also accounts for the high energy demand of the reaction. Consequently, the reductive elimination pathway for MetH-catalyzed methyl transfer from MeCbl to Hcy cannot be considered as one of the possible mechanistic routes.


Aristolochic acid I aggravates oxidative stress-mediated apoptosis by inhibiting APE1/Nrf2/HO-1 signaling.
Nephrotoxicity induced by aristolochic acid I (AAI) is related to redox stress and apoptosis. Apurinic/apyrimidine endonuclease 1 (APE1) has antioxidant and anti-apoptotic effects. This study investigated the potential role of APE1 in AAI-induced nephrotoxicity. Renal injury was successfully induced in C57BL/6J mice by intraperitoneal injection of AAI every other day for 28 days. Expressions of APE1, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase 1 (HO-1) in renal tissues of the model mice was inhibited, accompanied by oxidative damage and apoptosis. Similar results were obtained in vitro in human proximal tubular (HK-2) cells damaged by AAI. In the presence of a low concentration of the APE1 inhibitor E3330, expression of Nrf2 and HO-1 proteins in HK-2 cells was decreased and AAI-induced apoptosis was aggravated. Overexpression of APE1 in HK-2 cells promoted the expression of Nrf2 and HO-1, and alleviated apoptosis and renal injury induced by AAI. The collective findings demonstrate that AAI can inhibit the induction of oxidative stress and apoptosis by the APE1/Nrf2/HO-1 axis, leading to AAI renal injury. Targeting APE1 may be an effective therapeutic strategy to treat AA nephrotoxicity.


The cohesin complex regulates immunoglobulin class switch recombination.
Immunoglobulin (Ig) class switch recombination (CSR) is initiated by the transcription-coupled recruitment of activation-induced cytidine deaminase (AID) to switch regions and by the subsequent generation of double-stranded DNA breaks (DSBs). These DNA breaks are ultimately resolved through the nonhomologous end joining (NHEJ) pathway. We show that during CSR, AID associates with subunits of cohesin, a complex previously implicated in sister chromatid cohesion, DNA repair, and the formation of DNA loops between enhancers and promoters. Furthermore, we implicate the cohesin complex in the mechanism of CSR by showing that cohesin is dynamically recruited to the Sμ-Cμ region of the IgH locus during CSR and that knockdown of cohesin or its regulatory subunits results in impaired CSR and increased usage of microhomology-based end joining.