IL1R1+ cancer-associated fibroblasts drive tumor development and immunosuppression in colorectal cancer.

Fibroblasts have a considerable functional and molecular heterogeneity and can play various roles in the tumor microenvironment. Here we identify a pro-tumorigenic IL1R1+, IL-1-high-signaling subtype of fibroblasts, using multiple colorectal cancer (CRC) patient single cell sequencing datasets. This subtype of fibroblasts is linked to T cell and macrophage suppression and leads to increased cancer cell growth in 3D co-culture assays. Furthermore, both a fibroblast-specific IL1R1 knockout and IL-1 receptor antagonist Anakinra administration reduce tumor growth in vivo. This is accompanied by reduced intratumoral Th17 cell infiltration. Accordingly, CRC patients who present with IL1R1-expressing cancer-associated-fibroblasts (CAFs), also display elevated levels of immune exhaustion markers, as well as an increased Th17 score and an overall worse survival. Altogether, this study underlines the therapeutic value of targeting IL1R1-expressing CAFs in the context of CRC.

PexLoc-Parallel excitation using local encoding magnetic fields with nonlinear and nonbijective spatial profiles.

With the recent proposal of using magnetic fields that are nonlinear by design for spatial encoding, new flexibility has been introduced to MR imaging. The new degrees of freedom in shaping the spatially encoding magnetic fields (SEMs) can be used to locally adapt the imaging resolution to features of the imaged object, e.g., anatomical structures, to reduce peripheral nerve stimulation during in vivo experiments or to increase the gradient switching speed by reducing the inductance of the coils producing the SEMs and thus accelerate the imaging process. In this work, the potential of nonlinear and nonbijective SEMs for spatial encoding during transmission in multidimensional spatially selective excitation is explored. Methods for multidimensional spatially selective excitation radiofrequency pulse design based on nonlinear encoding fields are introduced, and it is shown how encoding ambiguities can be resolved using parallel transmission. In simulations and phantom experiments, the feasibility of selective excitation using nonlinear, nonbijective SEMs is demonstrated, and it is shown that the spatial resolution with which the target distribution of the transverse magnetization can be realized varies locally. Thus, the resolution of the target pattern can be increased in some regions compared with conventional linear encoding. Furthermore, experimental proof of principle of accelerated two-dimensional spatially selective excitation using nonlinear SEMs is provided in this study.

CYP98A3 from Arabidopsis thaliana is a 3'-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway.

The 4- and 5-hydroxylations of phenolic compounds in plants are catalyzed by cytochrome P450 enzymes. The 3-hydroxylation step leading to the formation of caffeic acid from p-coumaric acid remained elusive, however, alternatively described as a phenol oxidase, a dioxygenase, or a P450 enzyme, with no decisive evidence for the involvement of any in the reaction in planta. In this study, we show that the gene encoding CYP98A3, which was the best possible P450 candidate for a 3-hydroxylase in the Arabidopsis genome, is highly expressed in inflorescence stems and wounded tissues. Recombinant CYP98A3 expressed in yeast did not metabolize free p-coumaric acid or its glucose or CoA esters, p-coumaraldehyde, or p-coumaryl alcohol, but very actively converted the 5-O-shikimate and 5-O-d-quinate esters of trans-p-coumaric acid into the corresponding caffeic acid conjugates. The shikimate ester was converted four times faster than the quinate derivative. Antibodies directed against recombinant CYP98A3 specifically revealed differentiating vascular tissues in stem and root. Taken together, these data show that CYP98A3 catalyzes the synthesis of chlorogenic acid and very likely also the 3-hydroxylation of lignin monomers. This hydroxylation occurs on depsides, the function of which was so far not understood, revealing an additional and unexpected level of networking in lignin biosynthesis.

Cloning of Arabidopsis thaliana glutathione synthetase (GSH2) by functional complementation of a yeast gsh2 mutant.

Glutathione (L-gamma-glutamyl-L-cysteinylglycine, GSH) plays an important role in the protection of plants against various types of stress caused by reactive oxygen species, gazeous pollutants, heavy metals and xenobiotics. A cDNA fragment containing the entire coding unit for glutathione synthetase (GSH2) of Arabidopsis thaliana was cloned by complementation of the methylglyoxal sensitivity of a gsh2 mutant of the yeast Saccharomyces cerevisiae. The cDNA encodes a protein of 478 amino acids (deduced Mr: 53783), bearing clear sequence similarities to GSH2 products from frog embryos (Xenopus laevis), rat kidney (Rattus norvegicus) and from the fission yeast (Schizosaccharomyces pombe). A highly conserved glycine-rich domain close to the carboxy-terminus was found in the GSH2 product and appears to be typical for eukaryotic glutathione synthetases. The Mr is similar to those of soluble animal enzymes, suggesting that the Arabidopsis gene also codes for a cytosolic protein. Genomic DNA-blot analysis indicates the presence of a single GSH2 gene. The yeast gsh2 mutant becomes resistant to methylglyoxal and cadmium after transformation with the plasmid bearing the Arabidopsis GSH2 cDNA. Moreover, this increased resistance is correlated to the restoration of GSH content from below detectability in mutants to about 50% of the wild-type levels in transformed cells.

UDP-glucose sterol beta-D-glucosyltransferase, a plasma membrane-bound enzyme of plants: enzymatic properties and lipid dependence.

UDP-glucose sterol beta-D-glucosyltransferase (UDPG-SGTase) catalyzes the glucosylation of plant sterols. This enzyme has been shown to be membrane-bound, most of its activity being associated with plasma membrane in etiolated maize coleoptiles. After solubilization with detergents, total delipidation and purification, kinetic studies performed with a purified enzyme preparation in the presence of detergent and soybean phosphatidylcholine (PC) strongly suggest an ordered bi-bi mechanism for the glucosylation of sterols. A reduced sulfhydryl group and an arginyl residue were shown to be essential for activity. Lipid dependence studies have been performed on the delipidated enzyme in two systems: a micellar one composed of a mixture of enzyme, detergent and phospholipids and another one where the enzymatic activity was reconstituted in unilamellar lipid vesicles. In both systems it was shown that the UDPG-SGTase activity was stimulated to a large extent by negatively charged phospholipids. Enzymatic assays were performed with membrane fractions originating from plants whose sterol content was profoundly modified following treatment with a sterol biosynthesis inhibitor. Results showed that the sterol glucosylating activity was strongly inhibited in these fractions in accordance with sterol substrate specificity studies. All these results show that the UDPG-SGTase is exquisitely sensitive to its lipid environment. Physiological implications of these data are discussed in the light of the putative role of sterols in the plant cell.

[The effectiveness of electromyostimulation following operations of the capsule-ligament system of the knee joint in athletes]. / Wirksamkeit der Elektromyostimulation (EMS) nach Operationen am Kapsel-Band-Apparat des Kniegelenkes bei Sportlern.

The most important factor in rehabilitation after knee joint surgery is the influencing of the atrophy of the femoral musculature and the recovery of the arthromuscular balance. Electromyostimulation (EMS) should be an additional possibility of treatment. Longitudinal section studies were performed over 6 months in a group of 45 patients after cruciate ligament surgery and for a period of 3 months after meniscus surgery in a group of 61 patients, in each case with different aftertreatment programmes (with EMS vs without EMS). Muscle function control methods were used to assess the condition of the status of function of the musculature (M. vastus medialis). The effect achieved by EMS can be objectivated especially by means of the muscular tone parameters "elastic retraction" and "total compressibility". In the group of sportsmen with EMS treatment (low frequency constant frequency programme with mini-stimulant twice daily 15 minutes) these parameters are about 30% above those of the group without EMS. Timely and on-target use of EMS after knee joint surgery yields a better starting level for rehabilitation and is in our opinion mandatory as a matter of principle especially in high-performance sportsmen.

Cyclopropyl sterol and phospholipid composition of membrane fractions from maize roots treated with fenpropimorph.

Maize (Zea mays L.) caryopses were grown in the presence of fenpropimorph, a systemic fungicide, for 7 days in the dark. Membrane fractions enriched, respectively, in endoplasmic reticulum, plasma membrane, and mitochondria were isolated from control and treated maize roots and analyzed for their free sterol, phospholipid, and fatty acid composition. In treated plants, the intracellular distribution of free sterols was dramatically modified both qualitatively and quantitatively. The normally occurring Delta(5)-sterols disappeared almost completely and were replaced by 9beta, 19-cyclopropyl sterols, mainly cycloeucalenol and 24-methyl pollinastanol. These new compounds were found to accumulate in all the membrane fractions in such a way that the endoplasmic reticulum-rich fraction became the richest one in free sterols instead of the plasma membrane. In contrast, the fenpropimorph treatment of maize roots was shown not to affect either the relative proportions or the amounts of the individual phospholipids, but an increase in the unsaturation index of phospholipid-fatty acyl chains of the endoplasmic reticulum-rich fraction was observed. The present data suggest that, in higher plant membranes, cyclopropyl sterols could play a structural role similar to that of the bulk of Delta(5)-sterols.

[Objective assessment of the reactive effectiveness of middle frequency electromyostimulation using mechanical muscle diagnosis (myomechanography)]. / Mitteilung zur Objektivierung der reaktiven Wirksamkeit mittelfrequenter Elektromyostimulation mit Hilfe muskelmechanischer Diagnostik (Myomechanographie).

Using a new method of measuring passive-mechanical tonus-parameters of muscle tissue in situ ("Myomechanographie, MMG") we evaluated in 12 healthy men reactive changes of the muscle tonus immediate after one middle-frequent Electromyostimulations (EMS)-period (duration: 15 min). With the MMG was proved, that the applied EMS cause an organic effectiveness; the muscle in situ is more "elastic". This showes especially the increase of the single parameters 'elastic retraction' and 'total compressibility'. In the comparison group (n = 12) we obtained contrary after relaxly steady position (duration: 15 min.) clear reduced single parameters. The muscle is reactive less "elastic".

Regulation by Phospholipids and Kinetic Studies of Plant Membrane-Bound UDP-Glucose Sterol beta-d-Glucosyl Transferase.

Solubilization and partial purification of the microsomal UDP-glucose sterol glucosyl transferase activity from maize coleoptiles by chromatography on DEAE-cellulose resulted in a highly delipidated (>95%) and inactive enzymic preparation. Addition of sterols revealed part of the activity and subsequent addition of phospholipids further increased the activity. Negatively charged phospholipids were shown to be by far the best activators. The purification step also produced the elimination of two interfering microsomal enzymic activities: UDPase and steryl glucoside acyl transferase. The removal of these two enzymic activities was a prerequisite for kinetic studies including product-inhibition studies, since the substrates of these two latter enzymes are the products of UDPG-SGTase activity. The results of the kinetic studies strongly suggest an ordered bi-bi mechanism for the glucosylation of sterols. Finally the effect of different phospholipids on the kinetic parameters of the reaction was studied. Both phosphatidylcholine and phosphatidylglycerol significantly decrease K(m-sterol) (and not K(m-UDPglucose)) and increase the reaction V(max). The decrease of K(m-sterol) is similar with both phospholipids whereas the increase of V(max) is much greater with phosphatidylglycerol than with phosphatidylcholine.