CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells.

The prototypical genetic autoimmune disease is immune dysregulation polyendocrinopathy enteropathy X-linked (IPEX) syndrome, a severe pediatric disease with limited treatment options. IPEX syndrome is caused by mutations in the forkhead box protein 3 (FOXP3) gene, which plays a critical role in immune regulation. As a monogenic disease, IPEX is an ideal candidate for a therapeutic approach in which autologous hematopoietic stem and progenitor (HSPC) cells or T cells are gene edited ex vivo and reinfused. Here, we describe a CRISPR-based gene correction permitting regulated expression of FOXP3 protein. We demonstrate that gene editing preserves HSPC differentiation potential, and that edited regulatory and effector T cells maintain their in vitro phenotype and function. Additionally, we show that this strategy is suitable for IPEX patient cells with diverse mutations. These results demonstrate the feasibility of gene correction, which will be instrumental for the development of therapeutic approaches for other genetic autoimmune diseases.

Ha-ras(val12) induces HSP70b transcription via the HSE/HSF1 system, but HSP70b expression is suppressed in Ha-ras(val12)-transformed cells.

Heat shock proteins (Hsps) are overexpressed in many tumors, but are downregulated in some tumors. To check for a direct effect of Ha-Ras(val12) on HSP70 transcription, we transiently expressed the oncoprotein in Rat1 fibroblasts and monitored its effect on HSP70b promoter-driven reporter gene. We show that expression of Ha-Ras(val12) induced this promoter. Promoter analysis via systematic deletions and point mutations revealed that Ha-Ras(val12) induces HSP70b transcription via heat shock elements (HSEs). Also, Ha-Ras(val12) induction of HSE-mediated transcription was dramatically reduced in HSF1-/- cells. Yet, residual effect of Ha-Ras(val12) that was still measured in HSF1-/- cells suggests that some of the Ha-Ras(val12) effect is Hsf1-independent. When HSF1-/- cells, stably expressing Ha-Ras(val12), were grown on soft agar only small colonies were formed suggesting a role for heat shock factor 1 (Hsf1) in Ha-Ras(val12)-mediated transformation. Although Ha-ras(Val12) seems to be an inducer of HSP70's expression, we found that in Ha-ras(Val12-)transformed fibroblasts expression of this gene is suppressed. This suppression is correlated with higher sensitivity of Ha-ras(val12)-transformed cells to heat shock. We suggest that Ha-ras(Val12) is involved in Hsf1 activation, thereby inducing the cellular protective response. Cells that repress this response are perhaps those that acquire the capability to further proliferate and become transformed clones.

Time-resolved ARPES with sub-15 fs temporal and near Fourier-limited spectral resolution.

An experimental setup for time- and angle-resolved photoelectron spectroscopy with sub-15 fs temporal resolution is presented. A hollow-fiber compressor is used for the generation of 6.5 fs white light pump pulses, and a high-harmonic-generation source delivers 11 fs probe pulses at a photon energy of 22.1 eV. A value of 13 fs full width at half-maximum of the pump-probe cross correlation signal is determined by analyzing a photoemission intensity transient probing a near-infrared interband transition in 1T-TiSe2. Notably, the energy resolution of the setup conforms to typical values reported in conventional time-resolved photoemission studies using high harmonics, and an ultimate resolution of 170 meV is feasible.

Granzymes in age-related cardiovascular and pulmonary diseases.

Chronic inflammation is a hallmark of age-related cardiovascular and pulmonary diseases. Granzymes are a family of serine proteases that have been traditionally viewed as initiators of immune-mediated cell death. However, recent findings suggest that the pathophysiological role of granzymes is complex. Emerging functions for granzymes in extracellular matrix degradation, autoimmunity, and inflammation suggests a multifactorial mechanism by which these enzymes are capable of mediating tissue damage. Recent discoveries showing that granzymes can be produced and secreted by nonimmune cells during disease provide an additional layer of intricacy. This review examines the emerging biochemical and clinical evidence pertaining to intracellular and/or extracellular granzymes in the pathogenesis of aging and cardiopulmonary diseases.

[Quantitative evaluation of neurovegetative systemic changes based on the degree of minute-rhythmic coupling and control quality in acute and chronic diazépam therapy of neuroses]. / Quantitative Bewertung von neurovegetativen Systemveränderungen anhand des minutenrhythmischen Kopplungsgrades und der Regelgüte während akuter und chronischer Diazepamtherapie bei neurotischen Patienten

In 10 patients with definite forms of neurosis, controlphysiologic and biorhythmometric investigations were carried out to establish the effectiveness of diazepam therapy. After acute application of 10 mg diazepam, an increase in the degree of minute-rhythmic coupling correlated positively with a decrease of the control area, of the time adjustment of the heart rate after load-related deflection, and with an increase in a derived complex parameter of control quality. Chronic therapy with diazepam reversed the positive tendency of the biorhythmometric and control parameters. The results permit the conclusion that the minute-rhythmic coupling degree lends itself to diagnostic evaluation of the actual synchronization state or the related neurovegetative reaction state.