Childhood interleukin-6, C-reactive protein and atopic disorders as risk factors for hypomanic symptoms in young adulthood: a longitudinal birth cohort study

$\textbf{Background.}$ There are no existing longitudinal studies of inflammatory markers and atopic disorders in childhood and risk of hypomanic symptoms in adulthood. This study examined if childhood: (1) serum interleukin-6 (IL-6) and C-reactive protein (CRP); and (2) asthma and/or eczema are associated with features of hypomania in young adulthood. $\textbf{Method.}$ Participants in the Avon Longitudinal Study of Parents and Children, a prospective general population UK birth cohort, had non-fasting blood samples for IL-6 and CRP measurement at the age of 9 years ($n$ = 4645), and parents answered a question about doctor-diagnosed atopic illness before the age of 10 years ($n$ = 7809). These participants completed the Hypomania Checklist at age 22 years ($n$ = 3361). $\textbf{Results.}$ After adjusting for age, sex, ethnicity, socio-economic status, past psychological and behavioural problems, body mass index and maternal postnatal depression, participants in the top third of IL-6 values at 9 years, compared with the bottom third, had an increased risk of hypomanic symptoms by age 22 years [adjusted odds ratio 1.77, 95% confidence interval (CI) 1.10-2.85, $p$ < 0.001]. Higher IL-6 levels in childhood were associated with adult hypomania features in a dose-response fashion. After further adjustment for depression at the age of 18 years this association remained (adjusted odds ratio 1.70, 95% CI 1.03-2.81, $p$ = 0.038). There was no evidence of an association of hypomanic symptoms with CRP levels, asthma or eczema in childhood. $\textbf{Conclusions.}$ Higher levels of systemic inflammatory marker IL-6 in childhood were associated with hypomanic symptoms in young adulthood, suggesting that inflammation may play a role in the pathophysiology of mania. Inflammatory pathways may be suitable targets for the prevention and intervention for bipolar disorder.The UK Medical Research Council and the Wellcome Trust (grant reference 102215/2/13/2) and the University of Bristol provide core support for ALSPAC. J.F.H. is supported by a Medical Research Council Population Health Scientist Fellowship (grant code: MR/K021362/1)

Further Characterisation of the Molecular Signature of Quiescent and Activated Mouse Muscle Satellite Cells

Satellite cells are the resident stem cells of adult skeletal muscle. To date though, there is a paucity of native markers that can be used to easily identify quiescent satellite cells, with Pax7 probably being the best that is currently available. Here we have further characterized a number of recently described satellite cell markers, and also describe novel ones. Caveolin-1, integrin α7 and the calcitonin receptor proved reliable markers for quiescent satellite cells, being expressed by all satellite cells identified with Pax7. These three markers remained expressed as satellite cells were activated and underwent proliferation. The nuclear envelope proteins lamin A/C and emerin, mutations in which underlie Emery-Dreifuss muscular dystrophy, were also expressed in both quiescent and proliferating satellite cells. Conversely, Jagged-1, a Notch ligand, was not expressed in quiescent satellite cells but was induced upon activation. These findings further contribute to defining the molecular signature of muscle satellite cells

Integrated Functions of Pax3 and Pax7 in the Regulation of Proliferation, Cell Size and Myogenic Differentiation

Pax3 and Pax7 are paired-box transcription factors with roles in developmental and adult regenerative myogenesis. Pax3 and Pax7 are expressed by postnatal satellite cells or their progeny but are down regulated during myogenic differentiation. We now show that constitutive expression of Pax3 or Pax7 in either satellite cells or C2C12 myoblasts results in an increased proliferative rate and decreased cell size. Conversely, expression of dominant-negative constructs leads to slowing of cell division, a dramatic increase in cell size and altered morphology. Similarly to the effects of Pax7, retroviral expression of Pax3 increases levels of Myf5 mRNA and MyoD protein, but does not result in sustained inhibition of myogenic differentiation. However, expression of Pax3 or Pax7 dominant-negative constructs inhibits expression of Myf5, MyoD and myogenin, and prevents differentiation from proceeding. In fibroblasts, expression of Pax3 or Pax7, or dominant-negative inhibition of these factors, reproduce the effects on cell size, morphology and proliferation seen in myoblasts. Our results show that in muscle progenitor cells, Pax3 and Pax7 function to maintain expression of myogenic regulatory factors, and promote population expansion, but are also required for myogenic differentiation to proceed

Die Surfactantkonversion als enzymatischer Prozeß : Ist das Surfactantprotein SP-B ein Substrat der Konvertase?

Das in der Alveole der Säugerlungen vorkommende Surfactantmaterial kann in sogenannte small und large surfactant aggregates aufgetrennt werden. Zu den large surfactant aggregates zählen Lamellarkörperchen und tubuläres Myelin, also die biophysikalisch hochaktiven Präkursoren des interfacialen Surfactantfilms. Unter den Prämissen einer akuten respiratorischen Insuffizienz ist wiederholt festgestellt worden, dass die Verteilung zwischen den large surfactant aggregates und small surfactant aggregates sehr zugunsten der small surfactant aggregates verschoben ist. Hieraus resultierend findet sich ein Übergewicht dieser, biophysikalisch weitgehend inaktiven, Abbauprodukte des Grenzflächenfilms. Vor diesem Hintergrund wurde in der vorliegenden Doktorarbeit der Fragestellung nachgegangen, wodurch die alveoläre Umwandlung der large in die small surfactant aggregates, ein als Surfactantkonversion bezeichneter Vorgang, vermittelt wird, und ob diese Surfactantkonversion ein enzymatisch getriggerter Prozess ist. Zur Beantwortung dieser Frage wurde als Ausgangsmaterial eine gepoolte bronchoalveoläre Lavage von gesunden Kaninchen, sowie ein rekonstituiertes Surfactantmaterial verwendet. Methodisch kamen weiterhin chromatographische, elektrophoretische, biophysikalische Verfahren, sowie Enzymaktivitäts-Assays zur Anwendung. Zunächst einmal konnte festgestellt werden, dass für die weitreichende Konversion von Surfactant in vitro in der Tat die Gegenwart einer Esterase notwendig ist. Weiterhin ergab sich im Rahmen der Rekonstitutionsversuche mit variablen Surfactant-Apoproteinen ebenfalls der Hinweis, dass vor allen Dingen der relative Gehalt an SP-B einen weitreichenden Einfluss auf den Konversionsgrad ausübt. Bei der Untersuchung der Herkunft der Esteraseaktivität in der BAL zeigte sich, dass im Überstand der resuspendierten Zellen der bronchoalveolären Lavage, wie auch im Zelllysat erhebliche Mengen an Esteraseaktivität nachweisbar waren. Weiterhin wurde festgestellt, dass unter den Bedingungen einer in vitro Konversion die Esteraseaktivität in den Subfraktionen alveolären Surfactans zeitabhängig abfiel. So war in den large surfactant aggregates 42 min nach Beginn der in vitro Konversion überhaupt keine Esteraseaktivität und nur noch etwa ein Viertel der Amidaseaktivität nachweisbar. Auf der Suche nach dem möglichen Substrat dieser Esterase wurde sowohl für die natürliche, wie auch für isoliert mit - an Sepharose gekoppelter - Esterase inkubiertem Surfactantprotein B der Nachweis erbracht, dass im Rahmen des Konversionsprozesses das dimere SP-B abgebaut und ein Spaltprodukt in einem Molekulargewichtsbereich von 11-14 kDa neu auftritt. Eine aminoterminale Sequenzierung dieses Spaltproduktes ergab zweifelsfrei den Nachweis eines Surfactantprotein B entstammenden Proteins und zwar des aminoterminalen Anteils des SP-B. Dieses Spaltprodukt konnte durch ein neu entwickeltes HPLC-Verfahren zur Auftrennung der hydrophoben Surfactantproteine aus der BAL weiter aufgereinigt werden. Zusammenfassend ergibt sich auf der Basis der hier vorliegenden Daten der Befund, dass die Umwandlung von large in small surfactant aggregates und der hiermit verbundene Verlust der Oberflächenaktivität nicht nur von der Größe der Oberflächenveränderung, sondern zudem von der Gegenwart einer enzymatischen Aktivität abhängig sind. Im Rahmen der hier durchgeführten Untersuchung konnte der Nachweis einer Esteraseaktivität sowohl in den Zellen der BAL, wie auch im zellfreien Überstand erbracht werden. Als mögliches Substrat dieser Aktivität konnte das Surfactantprotein B identifiziert werden, für welches das Auftreten eines 11-14 kDa großen Spaltproduktes einwandfrei belegt werden konnte. Aus der Kenntnis dieser Ergebnisse leiten sich mögliche neue Therapieoptionen für das Acute Respiratory Distress Syndrome, wie auch für den Ventilator Induced Lung Injury ab, bei denen Verschiebungen des alveolären Surfactantpools zugunsten der small surfactant aggregates wiederholt beschrieben worden sind.The alveolar surfactant pool can be separated into the \u27large surfactant aggregates\u27 (LSA) and the \u27small surfactant aggregates\u27 (SSA). The LSA, including lamellar bodies and tubular myelin, represent the biophysically highly active precursors of the interfacial surfactant film. Under cyclic area changes LSA are converted into the SSA (surfactant conversion). In contrast to LSA the SSA are clearly less surface active. Under clinical conditions of the acute respiratory distress syndrome, the balance of LSA to SSA is found to be switched in favour of SSA. Under these conditions, the alveolar surfactant pool predominantly consists of the largely inactive small surfactant aggregates, thus favouring impairment of gas exchange and lung function. Drawn against this background we aimed to elucidate the mechanisms of the conversion process. To answer this question pooled bronchoalveolar lavages of healthy rabbits and reconstituted surfactant preparations were subjected to repetitive surface area changes in vitro and extend of conversion was analysed. Besides chromatographic, electrophoretic and biophysical techniques, enzyme activity assays were applied for experimental investigations. It was found that an esterase activity is necessary for the induction of surfactant conversion under cyclic surface area changes. Experiments with various concentrations of the different surfactant proteins SP-A, SP-B or SP-C in reconstituted lipid mixtures revealed that only SP-B has a profound impact on the extent of in vitro conversion. Enzyme activity assays showed high esterase activity in complete cell suspensions of bronchoalveolar lavage and cell lysates. Under conditions of in vitro conversion, the esterase activity was found to decline in dependency of the incubation time, resulting in complete loss of esterase activity after 42 min of in vitro conversion. With emphasis on the potential role of surfactant proteins as substrates of esterase activity, we could show that in vitro conversion of BAL as well as incubation of isolated SP-B with sepharose linked esterase would result in a cleavage of dimeric SP-B and detection of a new protein band with a molecular range of 11-14 kilodalton. Amino terminal sequencing revealed that this protein truly represents a cleavage product of the amino terminal part of SP-B. Further purification of the cleavage product was performed by a new developed HPLC method for separation from other hydrophobic surfactant proteins and phospholipids. In summary the presented data support the conclusion that conversion of large surfactant aggregates to small surfactant aggregates not only depends on cyclic changes of the air-liquid interface, but also on the presence of an esterase activity. This esterase activity was detected in the cytosolic fraction of BAL cells, mostly alveolar macrophages. A SP-B cleavage product with a molecular range of 11-14 kilodalton was identified upon in vitro incubation of esterase with SP-B and after in vitro conversion of a rabbit BAL pool, suggesting that SP-B is a substrate of the alveolar esterase. These data may help to identify new molecular targets to treat acute respiratory distress syndrome and ventilator induced lung injury

Three-Dimensional Human iPSC-Derived Artificial Skeletal Muscles Model Muscular Dystrophies and Enable Multilineage Tissue Engineering

Generating human skeletal muscle models is instrumental for investigating muscle pathology and therapy. Here, we report the generation of three-dimensional (3D) artificial skeletal muscle tissue from human pluripotent stem cells, including induced pluripotent stem cells (iPSCs) from patients with Duchenne, limb-girdle, and congenital muscular dystrophies. 3D skeletal myogenic differentiation of pluripotent cells was induced within hydrogels under tension to provide myofiber alignment. Artificial muscles recapitulated characteristics of human skeletal muscle tissue and could be implanted into immunodeficient mice. Pathological cellular hallmarks of incurable forms of severe muscular dystrophy could be modeled with high fidelity using this 3D platform. Finally, we show generation of fully human iPSC-derived, complex, multilineage muscle models containing key isogenic cellular constituents of skeletal muscle, including vascular endothelial cells, pericytes, and motor neurons. These results lay the foundation for a human skeletal muscle organoid-like platform for disease modeling, regenerative medicine, and therapy development

Profiles of physical, emotional and psychosocial wellbeing in the Lothian birth cohort 1936

<p>Abstract</p> <p>Background</p> <p>Physical, emotional, and psychosocial wellbeing are important domains of function. The aims of this study were to explore the existence of separable groups among 70-year olds with scores representing physical function, perceived quality of life, and emotional wellbeing, and to characterise any resulting groups using demographic, personality, cognition, health and lifestyle variables.</p> <p>Methods</p> <p>We used latent class analysis (LCA) to identify possible groups.</p> <p>Results</p> <p>Results suggested there were 5 groups. These included High (n = 515, 47.2% of the sample), Average (n = 417, 38.3%), and Poor Wellbeing (n = 37, 3.4%) groups. The two other groups had contrasting patterns of wellbeing: one group scored relatively well on physical function, but low on emotional wellbeing (Good Fitness/ Low Spirits,n = 60, 5.5%), whereas the other group showed low physical function but relatively well emotional wellbeing (Low Fitness/Good Spirits, n = 62, 5.7%). Salient characteristics that distinguished all the groups included smoking and drinking behaviours, personality, and illness.</p> <p>Conclusions</p> <p>Despite there being some evidence of these groups, the results also support a largely one-dimensional construct of wellbeing in old age—for the domains assessed here—though with some evidence that some individuals have uneven profiles.</p

A whole-cell biosensor for the detection of gold

Geochemical exploration for gold (Au) is becoming increasingly important to the mining industry. Current processes for Au analyses require sampling materials to be taken from often remote localities. Samples are then transported to a laboratory equipped with suitable analytical facilities, such as Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) or Instrumental Neutron Activation Analysis (INAA). Determining the concentration of Au in samples may take several weeks, leading to long delays in exploration campaigns. Hence, a method for the on-site analysis of Au, such as a biosensor, will greatly benefit the exploration industry. The golTSB genes from Salmonella enterica serovar typhimurium are selectively induced by Au(I/III)-complexes. In the present study, the golTSB operon with a reporter gene, lacZ, was introduced into Escherichia coli. The induction of golTSB::lacZ with Au(I/III)-complexes was tested using a colorimetric β-galactosidase and an electrochemical assay. Measurements of the β-galactosidase activity for concentrations of both Au(I)- and Au(III)-complexes ranging from 0.1 to 5 µM (equivalent to 20 to 1000 ng g⁻¹ or parts-per-billion (ppb)) were accurately quantified. When testing the ability of the biosensor to detect Au(I/III)-complexes(aq) in the presence of other metal ions (Ag(I), Cu(II), Fe(III), Ni(II), Co(II), Zn, As(III), Pb(II), Sb(III) or Bi(III)), cross-reactivity was observed, i.e. the amount of Au measured was either under- or over-estimated. To assess if the biosensor would work with natural samples, soils with different physiochemical properties were spiked with Au-complexes. Subsequently, a selective extraction using 1 M thiosulfate was applied to extract the Au. The results showed that Au could be measured in these extracts with the same accuracy as ICP-MS (P<0.05). This demonstrates that by combining selective extraction with the biosensor system the concentration of Au can be accurately measured, down to a quantification limit of 20 ppb (0.1 µM) and a detection limit of 2 ppb (0.01 µM).Carla M. Zammit, Davide Quaranta, Shane Gibson, Anita J. Zaitouna, Christine Ta, Joël Brugger, Rebecca Y. Lai, Gregor Grass, Frank Reit

Quantification of the Frequency and Multiplicity of Infection of Respiratory- and Lymph Node–Resident Dendritic Cells During Influenza Virus Infection

Background: Previous studies have demonstrated that DC differentially regulate influenza A virus (IAV)–specific CD8 T cell responses in vivo during high and low dose IAV infections. Furthermore, in vitro infection of DC with IAV at low versus high multiplicities of infection (MOI) results in altered cytokine production and a reduced ability to prime naïve CD8 T cell responses. Flow cytometric detection of IAV proteins within DC, a commonly used method for detection of cellular IAV infection, does not distinguish between the direct infection of these cells or their uptake of viral proteins from dying epithelial cells. Methods/Principal Findings: We have developed a novel, sensitive, single-cell RT-PCR–based approach to assess the infection of respiratory DC (rDC) and lymph node (LN)-resident DC (LNDC) following high and low dose IAV infections. Our results show that, while a fraction of both rDC and LNDC contain viral mRNA following IAV infection, there is little correlation between the percentage of rDC containing viral mRNA and the initial IAV inoculum dose. Instead, increasing IAV inoculums correlate with augmented rDC MOI. Conclusion/Significance: Together, our results demonstrate a novel and sensitive method for the detection of direct IAV infection at the single-cell level and suggest that the previously described ability of DC to differentially regulate IAV-specific T cell responses during high and low dose IAV infections could relate to the MOI of rDC within the LN rather than th

Acute interaction between hydrocortisone and insulin alters the plasma metabolome in humans

With the aim of identifying biomarkers of glucocorticoid action and their relationship with biomarkers of insulin action, metabolomic profiling was carried out in plasma samples from twenty healthy men who were administered either a low or medium dose insulin infusion (n = 10 each group). In addition, all subjects were given metyrapone (to inhibit adrenal cortisol secretion) +/-hydrocortisone (HC) in a randomised crossover design to produce low, medium and high glucocorticoid levels. The clearest effects of insulin were to reduce plasma levels of the branched chain amino acids (BCAs) leucine/isoleucine and their deaminated metabolites, and lowered free fatty acids and acylcarnitines. The highest dose of hydrocortisone increased plasma BCAs in both insulin groups but increased free fatty acids only in the high insulin group, however hydrocortisone did not affect the levels of acyl carnitines in either group. The clearest interaction between HC and insulin was that hydrocortisone produced an elevation in levels of BCAs and their metabolites which were lowered by insulin. The direct modulation of BCAs by glucocorticoids and insulin may provide the basis for improved in vivo monitoring of glucocorticoid and insulin action

Prolonged Antigen Presentation Is Required for Optimal CD8+ T Cell Responses against Malaria Liver Stage Parasites

Immunization with irradiated sporozoites is currently the most effective vaccination strategy against liver stages of malaria parasites, yet the mechanisms underpinning the success of this approach are unknown. Here we show that the complete development of protective CD8+ T cell responses requires prolonged antigen presentation. Using TCR transgenic cells specific for the malaria circumsporozoite protein, a leading vaccine candidate, we found that sporozoite antigen persists for over 8 weeks after immunization—a remarkable finding since irradiated sporozoites are incapable of replication and do not differentiate beyond early liver stages. Persisting antigen was detected in lymphoid organs and depends on the presence of CD11c+ cells. Prolonged antigen presentation enhanced the magnitude of the CD8+ T cell response in a number of ways. Firstly, reducing the time primed CD8+ T cells were exposed to antigen in vivo severely reduced the final size of the developing memory population. Secondly, fully developed memory cells expanded in previously immunized mice but not when transferred to naïve animals. Finally, persisting antigen was able to prime naïve cells, including recent thymic emigrants, to become functional effector cells capable of eliminating parasites in the liver. Together these data show that the optimal development of protective CD8+ T cell immunity against malaria liver stages is dependent upon the prolonged presentation of sporozoite-derived antigen