A novel TBP-TAF complex on RNA Polymerase II-transcribed snRNA genes

Initiation of transcription of most human genes transcribed by RNA polymerase II (RNAP II) requires the formation of a preinitiation complex comprising TFIIA, B, D, E, F, H and RNAP II. The general transcription factor TFIID is composed of the TATA-binding protein and up to 13 TBP-associated factors. During transcription of snRNA genes, RNAP II does not appear to make the transition to long-range productive elongation, as happens during transcription of protein-coding genes. In addition, recognition of the snRNA gene-type specific 3′ box RNA processing element requires initiation from an snRNA gene promoter. These characteristics may, at least in part, be driven by factors recruited to the promoter. For example, differences in the complement of TAFs might result in differential recruitment of elongation and RNA processing factors. As precedent, it already has been shown that the promoters of some protein-coding genes do not recruit all the TAFs found in TFIID. Although TAF5 has been shown to be associated with RNAP II-transcribed snRNA genes, the full complement of TAFs associated with these genes has remained unclear. Here we show, using a ChIP and siRNA-mediated approach, that the TBP/TAF complex on snRNA genes differs from that found on protein-coding genes. Interestingly, the largest TAF, TAF1, and the core TAFs, TAF10 and TAF4, are not detected on snRNA genes. We propose that this snRNA gene-specific TAF subset plays a key role in gene type-specific control of expression

Socio-semiotics and the symbiosis of humans, horses, and objects in later Iron Age Britain

This is an Accepted Manuscript of an article published by Taylor & Francis in Archaeological Journal on 14/03/2018, available online: https://www.tandfonline.com/doi/full/10.1080/00665983.2018.1441105Using an approach derived from material culture studies and semiotics, this paper addresses possible relationships between humans and horses in the British Iron Age.Through a study of the dominance of horse imagery found on Iron Age British coinage, specifically the Western coinage traditionally attributed to the 'Dobunni', the author explores how it may reflect possible relationships between humans and horses and their personhood therein. Drawing on wider faunal and metalwork evidence it is argued that these coins could be interpreted as a manifestation of the complex perspectives surrounding a symbiotic relationship between humans and horses

Remote ischaemic preconditioning protects against cardiopulmonary bypass-induced tissue injury: a preclinical study

Objectives: To test the hypothesis that remote ischaemic preconditioning (rIPC) reduces injury after cardiopulmonary bypass (CPB). Design: Randomised study with an experimental model of CPB (3 h CPB with 2 h of cardioplegic arrest). Twelve 15 kg pigs were randomly assigned to control or rIPC before CPB and followed up for 6 h. Intervention: rIPC was induced by four 5 min cycles of lower limb ischaemia before CPB. Main outcome measures: Troponin I, glial protein S-100B, lactate concentrations, load-independent indices (conductance catheter) of systolic and diastolic function, and pulmonary resistance and compliance were measured before and for 6 h after CPB. Results: Troponin I increased after CPB in both groups but during reperfusion the rIPC group had lower concentrations than controls (mean area under the curve 257.3 (SEM 7.3) v 89.0 (11.6) ng?h/ml, p = 0.02). Lactate increased after CPB in both groups but during reperfusion the control group had significantly more prolonged hyperlactataemia (p = 0.04). S-100B did not differ between groups. Indices of ventricular function did not differ. There was a tendency to improved lung compliance (p = 0.07), and pulmonary resistance changed less in the rIPC than in the control group during reperfusion (p = 0.02). Subsequently, peak inspiratory pressure was lower (p = 0.001). Conclusion: rIPC significantly attenuated clinically relevant markers of myocardial and pulmonary injury after CPB. Transient limb ischaemia as an rIPC stimulus has potentially important clinical applications

Expression of Trichoderma reesei cellulases CBHI and EGI in Ashbya gossypii

To explore the potential of Ashbya gossypii as a host for the expression of recombinant proteins and to assess whether protein secretion would be more similar to the closely related Saccharomyces cerevisiae or to other filamentous fungi, endoglucanase I (EGI) and cellobiohydrolase I (CBHI) from the fungus Trichoderma reesei were successfully expressed in A. gossypii from plasmids containing the two micron sequences from S. cerevisiae, under the S. cerevisiae PGK1 promoter. The native signal sequences of EGI and CBHI were able to direct the secretion of EGI and CBHI into the culture medium in A. gossypii. Although CBHI activity was not detected using 4- methylumbelliferyl-β-D-lactoside as substrate, the protein was detected by Western blot using monoclonal antibodies. EGI activity was detectable, the specific activity being comparable to that produced by a similar EGI producing S. cerevisiae construct. More EGI was secreted than CBHI, or more active protein was produced. Partial characterization of CBHI and EGI expressed in A. gossypii revealed overglycosylation when compared with the native T. reesei proteins, but the glycosylation was less extensive than on cellulases expressed in S. cerevisiae.Fundação para a Ciência e a Tecnologia (FCT

Non-invasive assessment of ventricular force–frequency relations in the univentricular circulation by tissue Doppler echocardiography: a novel method of assessing myocardial performance in congenital heart disease

Objective: To describe the first clinical application of a novel tissue Doppler derived index of contractility, isovolumic acceleration (IVA), in the assessment of the ventricular myocardial force–frequency relation (FFR) in the univentricular heart (UVH). Design: Prospective study. Setting: Tertiary referral centre. Interventions: Non-invasive assessment of the myocardial FFR by tissue Doppler echocardiography during atrial pacing. Results: IVA was used to measure the FFR of the systemic ventricle in patients with structurally normal hearts and in patients with UVHs. Basal IVA of the normal hearts (mean (SD) 1.9 (0.3) m/s(2)) was significantly greater than that of UVHs in patients with a dominant right ventricle (RV) (1.0 (0.3) m/s(2)) or left ventricle (LV) (0.8 (0.7) m/s(2); p < 0.05 for both). Neither the absolute nor percentage change from basal to peak values of IVA with pacing differed between the three groups. Peak force developed by the normal LV was significantly greater than that of the UVH, dominant LV group but not different from that of the UVH, dominant RV group. Conclusion: Contractility at basal heart rate is depressed in patients with UVH compared with the normal LV. Analysis of ventricular FFRs exposes further differences in myocardial contractility. There is no evidence that contractile function of the dominant RV is inferior to that of the dominant LV over a physiological range of heart rates

Assessment of the relationship between cerebral and splanchnic oxygen saturations measured by near‐infrared spectroscopy and direct measurements of systemic haemodynamic variables and oxygen transport after the Norwood procedure

OBJECTIVES: To evaluate the clinical utility of near‐infrared spectroscopic (NIRS) monitoring of cerebral (Sco(2)) and splanchnic (Sso(2)) oxygen saturations for estimation of systemic oxygen transport after the Norwood procedure. METHODS: Sco(2) and Sso(2) were measured with NIRS cerebral and thoracolumbar probes (in humans). Respiratory mass spectrometry was used to measure systemic oxygen consumption (V̇o(2)). Arterial (Sao(2)), superior vena caval (Svo2) and pulmonary venous oxygen saturations were measured at 2 to 4 h intervals to derive pulmonary (Qp) and systemic blood flow (Qs), systemic oxygen delivery (Do(2)) and oxygen extraction ratio (ERo(2)). Mixed linear regression was used to test correlations. A study of 7 pigs after cardiopulmonary bypass (study 1) was followed by a study of 11 children after the Norwood procedure (study 2). RESULTS: Study 1. Sco(2) moderately correlated with Svo(2), mean arterial pressure, Qs, Do(2) and ERo(2) (slope 0.30, 0.64. 2.30, 0.017 and −32.5, p < 0.0001) but not with Sao(2), arterial oxygen pressure (Pao(2)), haemoglobin and V̇o(2). Study 2. Sco(2) correlated well with Svo(2), Sao(2), Pao(2) and mean arterial pressure (slope 0.43, 0.61, 0.99 and 0.52, p < 0.0001) but not with haemoglobin (slope 0.24, p > 0.05). Sco(2) correlated weakly with V̇o(2) (slope −0.07, p  =  0.05) and moderately with Qs, Do(2) and ERo(2) (slope 3.2, 0.03, −33.2, p < 0.0001). Sso(2) showed similar but weaker correlations. CONCLUSIONS: Sco(2) and Sso(2) may reflect the influence of haemodynamic variables and oxygen transport after the Norwood procedure. However, the interpretation of NIRS data, in terms of both absolute values and trends, is difficult to rely on clinically