A Modular Digital Twinning Framework for Safety Assurance of Collaborative Robotics.

Digital twins offer a unique opportunity to design, test, deploy, monitor, and control real-world robotic processes. In this paper we present a novel, modular digital twinning framework developed for the investigation of safety within collaborative robotic manufacturing processes. The modular architecture supports scalable representations of user-defined cyber-physical environments, and tools for safety analysis and control. This versatile research tool facilitates the creation of mixed environments of Digital Models, Digital Shadows, and Digital Twins, whilst standardising communication and physical system representation across different hardware platforms. The framework is demonstrated as applied to an industrial case-study focused on the safety assurance of a collaborative robotic manufacturing process. We describe the creation of a digital twin scenario, consisting of individual digital twins of entities in the manufacturing case study, and the application of a synthesised safety controller from our wider work. We show how the framework is able to provide adequate evidence to virtually assess safety claims made against the safety controller using a supporting validation module and testing strategy. The implementation, evidence and safety investigation is presented and discussed, raising exciting possibilities for the use of digital twins in robotic safety assurance.

An improved method for an efficient and easily accessible eukaryotic ribosome display technology.

Ribosome display is a powerful in vitro technology for the selection and directed evolution of proteins. However, this technology has so far been perceived as being technically challenging owing to comparatively difficult protocols and the absence of tailored commercial reagents, particularly when using prokaryotic cell-free expression systems. Eukaryotic ribosome display is potentially a more accessible alternative because of the availability of suitable commercial reagents, yet despite published protocols, this method has been less widely used. For eukaryotic ribosome display, a novel mechanism of mRNA recovery compared with that of the well-proven prokaryotic method has been proposed. We have examined the eukaryotic ribosome display process with the aims of investigating the proposed mechanism of sequence recovery and of identifying aspects of the protocol that may have lead to poor performance and therefore so far limited its use. We demonstrate that the proposed novel method is in fact mechanistically comparable to the prokaryotic method and we provide a step-by-step protocol for eukaryotic ribosome display that is 20-fold more efficient than current published methods. Our findings should increase the ease of operating ribosome display technology, making it more accessible to the scientific community.

Effects of tumour necrosis factor-alpha in the human forearm: blood flow and endothelin-1 release.

Increased circulating concentrations of tumour necrosis factor-alpha (TNF-alpha) are seen in several pathological conditions associated with vascular disease. TNF-alpha induces the synthesis of endothelin-1 (ET-1), a potent vasoconstictor, by the endothelium. However, there is profound vasodilatation in sepsis, where circulating levels of both ET-1 and TNF-alpha are elevated. The details of the interaction between ET-1 and TNF-alpha and the predominant resulting haemodynamic effect in healthy humans are unclear. The aim of the present study was to determine the effects of intra-arterial TNF-alpha on ET-1 spillover, vascular tone and endothelial function in the healthy human forearm. Brachial arterial and deep venous blood samples, forearm plasma flow measurements and blood flow responses to acetylcholine and sodium nitroprusside were obtained in six healthy subjects before and during a 6 h infusion of TNF-alpha into the brachial artery. Forearm blood flow was significantly greater than baseline during exposure to TNF-alpha [median (lower quartile, upper quartile): baseline, 2.6 (2.1, 2.8) ml.min(-1).100 ml(-1); TNF-alpha, 4.6 (4.5, 5.1) ml.min(-1).100 ml(-1); P <0.05]. The rate of release of ET-1 was significantly greater than baseline after 30 and 260 min of TNF-alpha infusion [median (lower quartile, upper quartile): baseline, 0.8 (0.6, 1.1) pg.min(-1).100 ml(-1); 30 min, 2.4 (1.9, 3.2) pg.min(-1).100 ml(-1); 260 min, 4.1 (3.1, 4.2) pg.min(-1).100 ml(-1); P <0.05]. The vasodilatory response to acetylcholine was diminished during TNF-alpha infusion, whereas the response to sodium nitroprusside remained unchanged. We thus demonstrate for the first time that local TNF-alpha increases ET-1 spillover from the human forearm and impairs endothelium-dependent vasodilatation. In spite of this action, TNF-alpha has a vasodilatory effect, resulting in an increase in forearm blood flow.

Inter-relationships among gonadotrophins, reproductive steroids and inhibin in freemartin ewes.

Freemartins are sterile XX/XY chimaeras that occur as a result of placental fusion between male and female fetuses during early pregnancy. Freemartins occur predominantly in cattle, although the prevalence of ovine freemartinism is increasing. In this study, the reproductive endocrinology of ovine freemartins was compared with that of normal sheep. Freemartins had significantly (P < 0.001) higher basal concentrations of LH and FSH than did normal ewes or rams, although the response of LH to GnRH (10 microg) was similar in freemartins, ewes and rams. Resting concentrations of oestradiol were similar in freemartins and ewes and were increased in both after eCG administration. Testosterone concentrations were higher in freemartins than in ewes, but were unresponsive to GnRH or eCG. Administration of 62.5 mg progesterone or 25 lg oestradiol twice a day for 3 days suppressed LH concentrations to baseline values in freemartins, ewes and rams. In ewes, 500 microg oestradiol administered twice a day caused preovulatory surges in LH concentrations, but suppressed LH in freemartins to baseline values. Thus, LH secretion can potentially be regulated in freemartins by gonadal steroids. FSH concentrations in freemartins were not suppressed by doses of inhibin that were effective in ewes and rams. Therefore, freemartins behave in part like castrated animals, as they have high basal concentrations of LH and FSH, which can be stimulated by GnRH and suppressed by gonadal steroids. Conversely, inhibin does not suppress FSH concentrations in freemartins, and freemartins have circulating concentrations of steroids intermediate between those of castrated and normal animals.

Endothelin-1 synthesis reduced by red wine.

Statistical evidence of reduced coronary heart disease in areas of high wine consumption has led to the widespread belief that wine affords a protective effect. Although moderate drinking of any alcohol helps to reduce the incidence of coronary heart disease, there is no clear evidence that red wine confers an additional benefit. Here we show that red wines strongly inhibit the synthesis of endothelin-1, a vasoactive peptide that is crucial in the development of coronary atherosclerosis. Our findings indicate that components specific to red wine may help to prevent coronary heart disease.

Effects of transforming growth factor-beta1 on renal extracellular matrix components and their regulating proteins.

Transforming growth factor-beta1 (TGF-beta1) is widely regarded as a potent fibrogenic renal growth factor. In cell culture, TGF-beta1 has been shown to increase various extracellular matrix (ECM) proteins and tissue inhibitors of metalloproteinases (TIMP), while decreasing matrix metalloproteinases (MMP), providing the optimum environment for progressive ECM accumulation. This study, which uses the isolated perfused rat kidney (IPRK), describes for the first time in a whole kidney preparation the action of TGF-beta1 on factors associated with ECM processing. This model allows the study of the intact rat kidney with physiologic cell-cell interactions in the absence of confounding systemic influences. Left kidneys were removed from male Wistar rats by a nonischemic technique and perfused with a sterile, apyrogenic, endotoxin-free perfusate, based on the plasma volume expander Hemaccel (polygeline), at constant pressure in a recirculating IPRK system. Kidneys were perfused for 1 h either with (n = 3) or without (n = 3) recombinant human TGF-beta1 (20 ng/ml). The effects of perfusion were controlled by comparison with the nonperfused contralateral kidney (n = 6). TGF-beta1 was measured in the perfusate and urine, at the start and end of the experiment using an enzyme-linked immunosorbent assay to its biologically active form. After perfusion, sections of the kidneys were analyzed for changes in mRNA by Northern blotting. Significant increases in mRNA for fibronectin (7.5-fold, P < 0.01), heparan sulfate proteoglycan core protein (53-fold, P < 0.001), laminin beta1 (12-fold, P < 0.001), collagen alpha1(IV) (17-fold, P < 0.001), collagen alpha1(III) (fourfold, P < 0.001), and MMP9 (twofold, P < 0.05) were observed after perfusion with TGF-beta1. Measurement of TIMP1, TIMP2, TIMP3, MMP1, and MMP2 mRNA demonstrated no detectable change, whereas determination of mRNA for tissue transglutaminase, an enzyme capable of cross-linking many ECM components, showed an eightfold increase (P < 0.01). This study suggests that in the IPRK and in the absence of other exogenous growth factors, TGF-beta1 selectively increases the synthesis of ECM and tissue transglutaminase without changes that would result in the reduction of ECM degradation.

Transglutaminase transcription and antigen translocation in experimental renal scarring.

It was recently demonstrated that renal tissue transglutaminase (tTg) protein and its catalytic product the epsilon(gamma-glutamyl) lysine protein cross-link are significantly increased in the subtotal (5/6) nephrectomy model (SNx) of renal fibrosis in rats. It was proposed that the enzyme had two important physiologic functions in disease development; one of stabilizing the increased extracellular matrix (ECM) by protein cross-linking, the other in a novel form of tubular cell death. This study, using the same rat SNx model, demonstrates first by Northern blotting that expression of tTg mRNA when compared with controls is increased by day 15 (+70% increase, P < 0.05), then rises steadily, peaking at day 90 (+391%, P < 0.01), and remains elevated at 120 d (+205%, P < 0.05) when compared with controls. In situ hybridization histochemistry demonstrated that the tubular cells were the major site of the additional tTg synthesis. Immunohistochemistry on cryostat sections revealed a sixfold increase (P < 0.001) in ECM-bound tTg antigen at 90-d post-SNx, whereas in situ transglutaminase activity demonstrated by the incorporation of fluorescein cadaverine into cryostat sections indicated a 750% increase (P < 0.001) on day 90 in SNx animals. This increased activity was extracellular and predominantly found in the peritubular region. These results indicate that increased tTg gene transcription by tubular cells underlies the major changes in renal tTg protein reported previously in SNx rats, and that the presence of the epsilon(gamma-glutamyl) lysine cross-links in the extracellular environment is the result of the extracellular action of tTg. These changes may be in response to tubular cell injury during the scarring process and are likely to contribute to the progressive expansion of the ECM in renal fibrosis.

Effect of timing and size of photoperiod change on plasma FSH concentration and the correlation between FSH and age at first egg in pullets.

1. ISA Brown pullets were transferred at 6, 9, 12, 15, 18 or 20.3 weeks of age from an 8 h photoperiod to an 8, 10, 13 or 16 h photoperiod. Plasma follicle stimulating hormone (FSH) concentration was measured at transfer at 7 and 14 d afterwards, and age at first egg (AFE) was recorded. 2. Plasma FSH concentration in pullets reared on constant 8 h photoperiods generally increased with age but with a trough at 12 weeks. Plasma FSH increased during the first 14 d of photostimulation to a significantly higher concentration, compared with constant 8 h controls, when the photoperiod was increased to 13 or 16 h at 9, 12 or 15 weeks; but for the increase from 8 h to 10 h photoperiods FSH was only significantly higher than controls when the change was made at 12 weeks. 3. The change in plasma FSH concentration 14 d after photostimulation was significantly correlated with mean AFE (reported in Lewis et al., 1997) and appears to be a better predictor of gonadal development than concurrent changes in plasma LH concentration previously reported (Lewis et al., 1994).

Effect of constant and of changing photoperiod on plasma LH and FSH concentrations and age at first egg in layer strains of domestic pullets.

1. ISA Brown pullets were transferred from 8 to 14 h or from 14 to 8 h photoperiods at 35 or 56 d of age. Controls were maintained on constant 8 or 14 h photoperiods from day 1. 2. Blood samples were obtained immediately before each daylength change and subsequently at 7 d intervals until 1st egg in the treated groups and at 70 d of age and then at 14 d intervals until 1st egg in the constant photoperiod controls. Plasma luteinising hormone (LH) and follicle stimulating hormone (FSH) concentrations were determined using homologous radioimmunoassays. 3. Prior to 16 weeks, LH was consistently higher in birds on constant 14 h photoperiods than in those on constant 8 h, but was down-regulated as birds approached maturity so that LH concentrations in the 2 groups were similar during the final 10 d before the first egg was laid. FISH concentrations rose steadily with age but with a tendency for concentrations to be higher in the 8 h than in the 14 h treatment. Birds on constant 8 h daylengths matured 18.3 d later than those on constant 14 h photoperiods. 4. A 6 h increment in photoperiod given at 35 d or 56 d, resulted in an increase in LH within 7 d in both cases. FSH concentration did not respond to an increase in photoperiod at 35 d but rose following the same increase at 56 d. This was associated with a 3-week advance in sexual maturity, whilst age at 1st egg in birds photostimulated at 35 d was similar to the age with a constant 14 h photoperiod. 5. LH concentration fell when photoperiod was reduced from 14 to 8 h at either 35 or 56 d and remained below the constant 8 h controls for many weeks before rising to a concentration not significantly different from other groups in the final 10 d before 1st egg. FSH concentrations in birds exposed to a decreased daylength at 35 d, although more oscillatory, were similar to the constant 8 h photoperiod controls. In birds exposed to the same decrease at 56 d, FSH concentration initially tumbled but was similar in the 2 groups during the latter stages of rearing; neither differed significantly from the constant daylength controls during the 60 d before 1st egg. Sexual maturity in both groups given a reduction in photoperiod was delayed by about 2 weeks compared with constant 8 h controls. 6. Change in FSH concentration following an increase in daylength was a better predictor of age at 1st egg than change in LH. However, FSH concentrations after 14 weeks of age were rather similar in short day and long day controls and in the 2 groups given reductions in photoperiod at 35 d and 56 d, despite differences of nearly 5 weeks in mean age at 1st egg amongst these 4 treatments.

Responses of prepubertal and mature rams to thyroidectomy.

Thyroidectomy of seasonally breeding birds and mammals prevents the return to a state of sexual quiescence at the end of the breeding season. In starlings, thyroidectomy also causes premature sexual maturity. In this study, the effect of thyroidectomy upon the time of sexual maturity of prepubertal (8 week-old) ram lambs was examined. Thyroidectomy of four prepubertal and six mature rams was performed early in the spring. These and sham-operated controls were maintained in ambient photoperiods (south-west England). Scrotal circumference and serum LH, FSH, prolactin and thyroxine were measured every 2 weeks. In both the prepubertal lambs and the mature rams, scrotal circumference increased significantly within 5 weeks of thyroidectomy. FSH concentrations increased significantly in the mature rams after thyroidectomy. The relatively high FSH concentrations of thyroidectomized animals at the start of the experiment were maintained, but the FSH concentrations of intact lambs decreased during the late spring. These results provide the first indication that the timing of puberty in seasonally breeding mammals is a thyroid-dependent phenomenon.