Exonuclease-enhanced prime editors.

Prime editing (PE) is a powerful gene-editing technique based on targeted gRNA-templated reverse transcription and integration of the de novo synthesized single-stranded DNA. To circumvent one of the main bottlenecks of the method, the competition of the reverse-transcribed 3' flap with the original 5' flap DNA, we generated an enhanced fluorescence-activated cell sorting reporter cell line to develop an exonuclease-enhanced PE strategy ('Exo-PE') composed of an improved PE complex and an aptamer-recruited DNA-exonuclease to remove the 5' original DNA flap. Exo-PE achieved better overall editing efficacy than the reference PE2 strategy for insertions ≥30 base pairs in several endogenous loci and cell lines while maintaining the high editing precision of PE2. By enabling the precise incorporation of larger insertions, Exo-PE complements the growing palette of different PE tools and spurs additional refinements of the PE machinery.

Erratum: Therapeutic Atmosphere in Psychotherapy Sessions. Int. J. Environ. Res. Public Health 2020, 17, 4105.

The authors wish to make the following correction to their paper [...].

Therapeutic Atmosphere in Psychotherapy Sessions.

There is uncertainty concerning what the active ingredients in psychotherapy are. The First Experimental Study of Transference interpretations (FEST) was a randomized controlled trial of the effects of transference work (TW) in psychodynamic psychotherapy. Women with low quality of object relations (QOR) showed a large positive effect of transference work, while men with high QOR showed a slight negative effect. The present study aimed to expand the knowledge from the FEST by investigating the therapeutic atmosphere with Structural Analysis of Social Behavior (SASB). Two-way ANOVAs were conducted to investigate differences between SASB cluster scores between subgroups. The therapeutic atmosphere was characterized by Protect-Trust, Affirm-Disclose and Control-Submit. Multilevel modeling was used to assess the relationship between a therapist variable and outcomes for men and women. Contrary to expectations, no significant differences in therapeutic atmosphere between subgroups (with or without TW in women with low QOR and men with high QOR) were observed using the process measure SASB.

Application-oriented fundamental research on concrete and reinforced concrete structures: selected findings from an Austro-Chinese research project.

In this paper, the significance of application-oriented fundamental research on concrete and reinforced concrete structures for progress regarding practical applications to structural design is addressed based on four examples. They were treated in a joint research project of Vienna University of Technology and Tongji University. The first topic refers to sudden heating or cooling of concrete structures, the second one to high-dynamic strength of specimens made of cementitious materials, the third one to structural analysis of segmental tunnel rings used in mechanized tunneling, and the fourth one to serviceability and ultimate limit states of concrete hinges used in integral bridge construction. The first two topics deal with exceptional load cases. Results from the fundamental research call for improvements of state-of-the-art simulation approaches used in civil engineering design. The last two topics refer to reinforced concrete hinges used in mechanized tunneling and integral bridge construction, respectively. Integrative research has led to progress regarding the verification of serviceability and ultimate limit states. In all four examples, results from fundamental research are used to scrutinize state-of-the-art approaches used in practical structural design of civil engineering structures. This allows for identifying interesting directions for the future development of design guidelines and standards.

Early-Age Evolution of Strength, Stiffness, and Non-Aging Creep of Concretes: Experimental Characterization and Correlation Analysis.

Six different concretes are characterized during material ages between 1 and 28 days. Standard tests regarding strength and stiffness are performed 1, 3, 7, 14, and 28 days after production. Innovative three-minute-long creep tests are repeated hourly during material ages between one and seven days. The results from the standard tests are used to assess and to improve formulas of the fib Model Code 2010: the correlation formula between the 28-day values of the strength and the stiffness, and the evolution formulas describing the early-age evolution of the strength and the stiffness during the first four weeks after production. The results from the innovative tests are used to develop a correlation formula between the 28-day values of Young's modulus and the creep modulus, and an evolution formula describing the early-age evolution of the creep modulus during the first four weeks after production. Particularly, the analyzed CEM I concretes develop stiffness and strength significantly faster than described by the formulas of the Model Code. The creep modulus of the investigated concretes evolves significantly slower than their strength and stiffness. Thus, concrete loaded at early ages is surprisingly creep active, even if the material appears to be quite mature in terms of its strength and stiffness.

The lymphocyte migration inhibitor FTY720 attenuates experimental hypertensive nephropathy.

The lymphocyte migration inhibitor FTY720 attenuates experimental hypertensive nephropathy. Infiltration with lymphocytes is found in both immune and nonimmune chronic kidney diseases. In a rat model of immune-initiated progressive glomerulosclerosis, selective inhibition of lymphocyte infiltration by FTY720 showed significant beneficial effects on renal fibrosis. To test whether this translates into hypertensive nephropathy (HN), the lymphocyte migration inhibitor was administered to rats following nephrectomy. Two days after surgery, male Wistar rats were allocated to the following groups: Sham surgery, nephrectomy (HN), and HN + FTY720 (0.3 mg/kg body wt). Therapy was continued for 6 wk. Treatment with FTY720 was found to selectively reduce blood lymphocyte counts by 85% (P < 0.001 vs. HN) and renal lymphocyte infiltration (CD-3 positive cells) by 63% (P < 0.01 vs. HN) as was anticipated. Lymphocyte depletion went along with a significant reduction in proteinuria (-28%), whereas hypertensive systemic blood pressure remained unchanged (160 +/- 5 vs. 161 +/- 5 mmHg, P = not significant). The markedly increased histological tubulointerstitial and glomerular matrix protein accumulation, collagen, laminin, and fibronectin deposition were all significantly impeded in the FTY720-treated animals. The anti-fibrotic effects of FTY720 were paralleled by significant reductions in renal transforming growth factor (TGF)-beta overexpression, macrophage infiltration, and cell proliferation. In conclusion, the lymphocyte migration inhibitor FTY720 significantly limits histological and molecular fibrosis in a model of hypertensive nephropathy without affecting increased systemic blood pressure. Prevention of renal lymphocytes' infiltration by FTY720 was followed by significant reductions in TGF-beta overexpression, macrophage infiltration, and renal cell proliferation. These results suggest that infiltrating lymphocytes play an active, profibrotic role in the progression of hypertensive renal tissue injury.

Low-dose mTOR inhibition by rapamycin attenuates progression in anti-thy1-induced chronic glomerulosclerosis of the rat.

Treatment options in human mesangioproliferative glomerulonephritis/sclerosis, mostly IgA nephropathy, are limited. Progressive mesangioproliferative nephropathy represents a major cause of end-stage kidney disease. The present study explores the efficacy of low-dose mTOR inhibition by rapamycin in a chronic-progressive model of mesangioproliferative glomerulosclerosis (cGS). cGS was induced by high-dose anti-thy1 antibody injection into uninephrectomized rats. Rapamycin administration (2.5 mg.kg(-1).body wt(-1)) was started 10 days after antibody injection and continued until week 20. cGS was characterized by advancing proteinuria, increased blood pressure, marked tubulointerstitial and glomerular fibrosis, cell proliferation and round cell infiltration, and impaired renal function. Kruskal-Wallis and Mann-Whitney U-tests were used for statistical analysis. The course of chronic anti-thy1-induced glomerulosclerosis was significantly attenuated by low-dose rapamycin treatment. In week 20, this was demonstrated by improvements in proteinuria (-38%), systolic blood pressure (-16 mmHg), tubulointerstitial and glomerular histological matrix accumulation (-61 and -24%), transforming growth factor-beta1 overexpression (-41 and -47%), collagen I deposition (-53 and -65%), cell proliferation (-90 and -76%), and leukocyte number (macrophages -52 and -53%; lymphocytes -58 and 51%), respectively. Rapamycin improved renal function as well (blood creatinine -0.68 mg/dl, urea -66.7 mg/day, and creatinine clearance +0.13 ml.min(-1).100 g body wt(-1)). In conclusion, low-dose mTOR inhibition by rapamycin limits the progressive course of anti-thy1-induced renal disease toward chronic glomerulosclerosis, tubulointerstitial fibrosis, and renal insufficiency. Renoprotection by rapamycin involved significant beneficial effects on multiple key pathways in the progression of chronic renal disease, i.e., proteinuria, extracellular matrix accumulation, renal cell proliferation, and inflammatory cell infiltration.

Microbiologicals for deactivating mycotoxins.

Mycotoxins are secondary metabolites of fungi affecting human and animal health. Five classes of mycotoxins are of major concern in animal husbandry, namely aflatoxins, trichothecenes, zearalenone, ochratoxins, and fumonisins. Due to their diverse structure these fungal toxins are able to cause a great variety of acute symptoms in animals. Clay minerals have been used in animal nutrition to bind mycotoxins, but the binders are only very specific for aflatoxins but not for other toxins. A novel strategy to control the problem of mycotoxicoses in animals is the application of microorganisms capable of biotransforming mycotoxins into nontoxic metabolites. The microbes act in the intestinal tract of animals prior to the resorption of the mycotoxins. A Eubacterium (BBSH 797) strain is able to deactivate trichothecenes by reduction of the epoxide ring (CAST, Mycotoxins, Risks in Plant, Animal and Human Systems, Task Force Report 139, Council of Agricultural Science and Technology, Ames Iowa 2003, p. 10.; Binder, E. M., Binder, J., Ellend, N., Schaffer, E. et al., in: Miraglia, M., van Egmond, H., Brera, C., Gilbert, J. (Eds.), Mycotoxins and Phycotoxins--Developments in Chemistry, Toxicology and Food Safety, Alaken, Fort Collins 1996, pp. 279-285). This strain was isolated out of bovine rumen fluid and the mode of action was proven in vitro and also in vivo. Further a novel yeast strain, capable of degrading ochratoxin A and zearalenone was isolated and characterized (Bruinink, A., Rasonyi, T., Sidler, C., Nat. Toxins 1999, 6, 173-177; Schatzmayr, G., Heidler, D., Fuchs, E., Mohnl, M. et al., Mycotoxin Res. 2003, 19, 124-128.) Due to the yeasts affiliation to the genus of Trichosporon and its property to degrade mycotoxins this strain was named Trichosporon mycotoxinivorans (Trichosporon MTV, 115).