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Sigma factors are transcriptional regulators that are part of complex regulatory networks for major cellular processes, as well as for growth phase-dependent regulation and stress response. Actinoplanes sp. SE50/110 is the natural producer of acarbose, an α-glucosidase inhibitor that is used in diabetes type 2 treatment. Acarbose biosynthesis is dependent on growth, making sigma factor engineering a promising tool for metabolic engineering. ACSP50_0507 is a homolog of the developmental and osmotic-stress-regulating Streptomyces coelicolor σHSc. Therefore, the protein encoded by ACSP50_0507 was named σHAs. Here, an Actinoplanes sp. SE50/110 expression strain for the alternative sigma factor gene ACSP50_0507 (sigHAs) achieved a two-fold increased acarbose yield with acarbose production extending into the stationary growth phase. Transcriptome sequencing revealed upregulation of acarbose biosynthesis genes during growth and at the late stationary growth phase. Genes that are transcriptionally activated by σHAs frequently code for secreted or membrane-associated proteins. This is also mirrored by the severely affected cell morphology, with hyperbranching, deformed and compartmentalized hyphae. The dehydrated cell morphology and upregulation of further genes point to a putative involvement in osmotic stress response, similar to its S. coelicolor homolog. The DNA-binding motif of σHAs was determined based on transcriptome sequencing data and shows high motif similarity to that of its homolog. The motif was confirmed by in vitro binding of recombinantly expressed σHAs to the upstream sequence of a strongly upregulated gene. Autoregulation of σHAs was observed, and binding to its own gene promoter region was also confirmed.

Abstract Background Canine mast cell tumour proliferation depends to a large extent on the activity of KIT, a tyrosine kinase receptor. Inhibitors of the KIT tyrosine kinase have recently been introduced and successfully applied as a therapeutic agent for this tumour type. However, little is known on the downstream target genes of this signaling pathway and molecular changes after inhibition. Results Transcriptome analysis of the canine mast cell tumour cell line C2 treated for up to 72 hours with the tyrosine kinase inhibitor masitinib identified significant changes in the expression levels of approximately 3500 genes or 16% of the canine genome. Approximately 40% of these genes had increased mRNA expression levels including genes associated with the pro-proliferative pathways of B- and T-cell receptors, chemokine receptors, steroid hormone receptors and EPO-, RAS and MAP kinase signaling. Proteome analysis of C2 cells treated for 72 hours identified 24 proteins with changed expression levels, most of which being involved in gene transcription, e.g. EIA3, EIA4, TARDBP, protein folding, e.g. HSP90, UCHL3, PDIA3 and protection from oxidative stress, GSTT3, SELENBP1. Conclusions Transcriptome and proteome analysis of neoplastic canine mast cells treated with masitinib confirmed the strong important and complex role of KIT in these cells. Approximately 16% of the total canine genome and thus the majority of the active genes were significantly transcriptionally regulated. Most of these changes were associated with reduced proliferation and metabolism of treated cells. Interestingly, several pro-proliferative pathways were up-regulated, which may represent attempts of masitinib treated cells to activate alternative pro-proliferative pathways. These pathways may contain hypothetical targets for a combination therapy with masitinib to further improve its therapeutic effect.

Magnetic nanofibers were prepared by electrospinning polymer/metal solutions of poly(acrylonitrile) (PAN) with magnetite (Fe3O4) nanoparticles. At a mass ratio of PAN:magnetite of 2:1, the total solid content in the dimethyl sulfoxide (DMSO) solution was varied between 15 wt.% and 25 wt.%, which represents the limits of the spinnable range. The results show that the most homogeneous nanofiber mats were 21 wt.% solid contents. At 15 wt.% solid contents, a nano-membrane with some fibrous regions was produced. Nanofibers at 25 wt.% had a much larger and more inhomogeneous diameter. Nevertheless, the magnetic properties of all samples were very similar, indicating that the distribution of magnetite nanoparticles in the fibers is comparable in all samples. The results also suggested that the samples spun from solutions with near-ideal solid contents (19–21 wt.%) contain agglomerations of the nanoparticles inside the nanofibers.

Large uncertainties in model parameterizations and input data sets make projections of future sea level rise contributions of outlet glaciers challenging. Here we introduce a novel technique for weighing large ensemble model simulations that uses information of key observables. The approach is robust to input errors and yields calibrated means and error estimates of a glacier's mass balance. We apply the technique to Jakobshavn Isbræ, using a model that includes a dynamic calving law, and closely reproduce the observed behavior from 1985 to 2018 by forcing the model with ocean temperatures only. Our calibrated projection suggests that the glacier will continue to retreat and contribute about 5.1 mm to eustatic sea level rise by 2100 under present-day climatic forcing. Our analysis shows that the glacier's future evolution will strongly depend on the ambient oceanic setting.

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Sigma factors are transcriptional regulators that are part of complex regulatory networks for major cellular processes, as well as for growth phase-dependent regulation and stress response. Actinoplanes sp. SE50/110 is the natural producer of acarbose, an α-glucosidase inhibitor that is used in diabetes type 2 treatment. Acarbose biosynthesis is dependent on growth, making sigma factor engineering a promising tool for metabolic engineering. ACSP50_0507 is a homolog of the developmental and osmotic-stress-regulating Streptomyces coelicolor σHSc. Therefore, the protein encoded by ACSP50_0507 was named σHAs. Here, an Actinoplanes sp. SE50/110 expression strain for the alternative sigma factor gene ACSP50_0507 (sigHAs) achieved a two-fold increased acarbose yield with acarbose production extending into the stationary growth phase. Transcriptome sequencing revealed upregulation of acarbose biosynthesis genes during growth and at the late stationary growth phase. Genes that are transcriptionally activated by σHAs frequently code for secreted or membrane-associated proteins. This is also mirrored by the severely affected cell morphology, with hyperbranching, deformed and compartmentalized hyphae. The dehydrated cell morphology and upregulation of further genes point to a putative involvement in osmotic stress response, similar to its S. coelicolor homolog. The DNA-binding motif of σHAs was determined based on transcriptome sequencing data and shows high motif similarity to that of its homolog. The motif was confirmed by in vitro binding of recombinantly expressed σHAs to the upstream sequence of a strongly upregulated gene. Autoregulation of σHAs was observed, and binding to its own gene promoter region was also confirmed. [ABSTRACT FROM AUTHOR]

Glacier-front dynamics is an important control on Greenland's ice mass balance. Warmer ocean waters trigger ice-front retreats of marine-terminating glaciers, and the corresponding loss in resistive stress leads to glacier acceleration and thinning. Here we present an approach to quantify the sensitivity and vulnerability of marine-terminating glaciers to ocean-induced melt. We develop a plan view model of Store Gletscher that includes a level set-based moving boundary capability, a parameterized ocean-induced melt, and a calving law with complete and precise land and fjord topographies to model the response of the glacier to increased melt. We find that the glacier is stabilized by a sill at its terminus. The glacier is dislodged from the sill when ocean-induced melt quadruples, at which point the glacier retreats irreversibly for 27 km into a reverse bed. The model suggests that ice-ocean interactions are the triggering mechanism of glacier retreat, but the bed controls its magnitude.

Calving is a major mechanism of ice discharge of the Antarctic and Greenland ice sheets, and a change in calving front position affects the entire stress regime of marine terminating glaciers. The representation of calving front dynamics in a 2-D or 3-D ice sheet model remains non-trivial. Here, we present the theoretical and technical framework for a level-set method, an implicit boundary tracking scheme, which we implement into the Ice Sheet System Model (ISSM). This scheme allows us to study the dynamic response of a drainage basin to user-defined calving rates. We apply the method to Jakobshavn Isbræ, a major marine terminating outlet glacier of the West Greenland Ice Sheet. The model robustly reproduces the high sensitivity of the glacier to calving, and we find that enhanced calving triggers significant acceleration of the ice stream. Upstream acceleration is sustained through a combination of mechanisms. However, both lateral stress and ice influx stabilize the ice stream. This study provides new insights into the ongoing changes occurring at Jakobshavn Isbræ and emphasizes that the incorporation of moving boundaries and dynamic lateral effects, not captured in flow-line models, is key for realistic model projections of sea level rise on centennial timescales.

The nature of the phase transition has been studied in MnNi _1− _x Co _x Ge _0.97 Al _0.03 ( x = 0.20–0.50) through magnetization, differential scanning calorimetry and x-ray diffraction measurements; and the associated reversibility in the magnetocaloric effect has been examined. A small amount of Al substitution for Ge can lower the structural phase transition temperature, resulting in a coupled first-order magnetostructural transition (MST) from a ferromagnetic orthorhombic to a paramagnetic hexagonal phase in MnNi _1− _x Co _x Ge _0.97 Al _0.03 . Interestingly, a composition-dependent triple point (TP) has been detected in the studied system, where the first-order MST is split into an additional phase boundary at higher temperature with a second-order transition character. The critical-field-value of the field-induced MST decreases with increasing Co concentration and disappears at the TP ( x = 0.37) resembling most field-sensitive MST among the studied compositions. An increase of the hexagonal lattice parameter a _hex near the TP indicates a lattice softening associated with an enhancement of the vibrational amplitude in the Ni/Co site. The lattice softening leads to a larger field-induced structural entropy change (structural entropy change≫ magnetic entropy change, for this class of materials) with the application of a lower field, which results in a larger reversibility of the low-field entropy change (|Δ S _rev | = 6.9 J kg ^−1 K for Δ μ _0 H = 2 T) at the TP.

Calving is a major mechanism of ice discharge of the Antarctic and Greenland ice sheets, and a change in calving front position affects the entire stress regime of marine terminating glaciers. The representation of calving front dynamics in a 2-D or 3-D ice sheet model remains non-trivial. Here, we present the theoretical and technical framework for a level-set method, an implicit boundary tracking scheme, which we implement into the Ice Sheet System Model (ISSM). This scheme allows us to study the dynamic response of a drainage basin to user-defined calving rates. We apply the method to Jakobshavn Isbræ, a major marine terminating outlet glacier of the West Greenland Ice Sheet. The model robustly reproduces the high sensitivity of the glacier to calving, and we find that enhanced calving triggers significant acceleration of the ice stream. Upstream acceleration is sustained through a combination of mechanisms. However, both lateral stress and ice influx stabilize the ice stream. This study provides new insights into the ongoing changes occurring at Jakobshavn Isbræ and emphasizes that the incorporation of moving boundaries and dynamic lateral effects, not captured in flow-line models, is key for realistic model projections of sea level rise on centennial timescales.

In this review article, we conceptually investigated the requirements of magnetic nanoparticles for their application in biosensing and related them to example systems of our thin-film portfolio. Analyzing intrinsic magnetic properties of different magnetic phases, the size range of the magnetic particles was determined, which is of potential interest for biosensor technology. Different e-beam lithography strategies are utilized to identify possible ways to realize small magnetic particles targeting this size range. Three different particle systems from 500 μm to 50 nm are produced for this purpose, aiming at tunable, vertically magnetized synthetic antiferromagnets, martensitic transformation in a single elliptical, disc-shaped Heusler Ni50Mn32.5Ga17.5 particle and nanocylinders of Co2MnSi-Heusler compound. Perspectively, new applications for these particle systems in combination with microfluidics are addressed. Using the concept of a magnetic on–off ratchet, the most suitable particle system of these three materials is validated with respect to magnetically-driven transport in a microfluidic channel. In addition, options are also discussed for improving the magnetic ratchet for larger particles.

The article presents Jehovah’s witnesses’ moral teaching on the subject of preparation for marital and family life. According to Jehovah’s Witnesses, proper preparation for a married life and family forecasts good prospects for the future, people in personality development and tread breeding of a moral and religious attitude. Man realizes himself when his relationship does not change. Duly conceived moral principles sensitize people to the problems they enter in fiancée relations. Issues regarding the selection of future reasonable, non-misplaced love. Jehovah’s Witnesses show the difference between infatuation, and the true love of the other person. In the modern era, many brides explain common sexual conditions and obligations as something normal. Paradoxually the sins of people’s connections. The elders of the congregation did not begin to drink in connection with another thing. In the pages of the Watchtower Society articles, the issue of the contraception problem leaves the objections. All behavior is unacceptable. The constant motif that is repeated in learning Jehovah’s Witnesses provides self-esteem for self-satisfaction and directing trials to the Creator. Only Jehovah’s intervention can help to preserve the patterns of moral life.

The aim of the present study was to elucidate the effects of the probiotic feed additive Enterococcus faecium NCIMB 10415 (E. faecium) on porcine jejunal epithelial cells (IPEC-J2) during an in vitro challenge with enterotoxigenic Escherichia coli (ETEC). Cells were incubated with E. faecium, ETEC, or both, and the effects on barrier function and structure and intra- and intercellular signaling were determined. Coincubation with E. faecium abolished the ETEC-induced decrease in transepithelial resistance (Rt) (p≤0.05). No differences were seen in the expression levels of the intercellular connecting tight junction proteins examined. However, for the first time, a reorganization of the monolayer was observed in ETEC-infected cells but not in coincubated cells. ETEC induced an increase in cytotoxicity that was prevented by coincubation (p≤0.05), whereas apoptosis rates were not affected by bacterial treatment. ETEC increased the mRNA expression and release of proinflammatory cytokines TNF-α, IL-1α, and IL-6 which could be prevented by coincubation for TNF-α mRNA expression and IL-6 protein (p≤0.05). Likewise, cAMP concentrations elevated by ETEC were reduced in coincubated cells (p≤0.05). These findings indicate a protective effect of the probiotic E. faecium on inflammatory responses during infection with ETEC.

In chronic respiratory disease, matrix metalloproteinases (MMPs) contribute to pathological tissue destruction when expressed in excess, while tissue inhibitors of metalloproteinases (TIMPs) counteract MMPs with overexpression leading to fibrosis formation. They may be out of balance in equine pneumopathies and serve as biomarkers of pulmonary inflammation. We hypothesized that MMPs and TIMPs correlate to clinical findings and bronchoalveolar lavage fluid cytology in different equine chronic pneumopathies. Using a scoring system, 61 horses were classified controls as free of respiratory disease (n=15), recurrent airway obstruction (RAO, n=17), inflammatory airway disease (IAD, n=18), or chronic interstitial pneumopathy (CIP, n=11). Zymography and equine MMP and TIMP assays were used to detect MMP-2, MMP-8, MMP-9 as well as TIMP-1, and TIMP-2 in BALF supernatant. MMP-2, TIMP-1, and TIMP-2 concentrations were significantly increased in RAO and IAD compared to controls. MMP-9 concentration and MMP-8 activity evaluated by fluorimetry were significantly increased in RAO, IAD, and CIP. These results were confirmed by zymography for MMP-2 and MMP-9 activity in 52 horses. In conclusion, MMPs and TIMPs correlate well with clinical and cytologic findings. These findings support the usefulness of MMPs, TIMPs, and their ratios to evaluate the severity of respiratory disease and may help to identify subclinical cases.