The nucleoside diphosphate kinase gene Nme3 acts as quantitative trait locus promoting non-Mendelian inheritance.

The t-haplotype, a variant form of the t-complex region on mouse chromosome 17, acts as selfish genetic element and is transmitted at high frequencies (> 95%) from heterozygous (t/+) males to their offspring. This phenotype is termed transmission ratio distortion (TRD) and is caused by the interaction of the t-complex responder (Tcr) with several quantitative trait loci (QTL), the t-complex distorters (Tcd1 to Tcd4), all located within the t-haplotype region. Current data suggest that the distorters collectively impair motility of all sperm derived from t/+ males; t-sperm is rescued by the responder, whereas (+)-sperm remains partially dysfunctional. Recently we have identified two distorters as regulators of RHO small G proteins. Here we show that the nucleoside diphosphate kinase gene Nme3 acts as a QTL on TRD. Reduction of the Nme3 dosage by gene targeting of the wild-type allele enhanced the transmission rate of the t-haplotype and phenocopied distorter function. Genetic and biochemical analysis showed that the t-allele of Nme3 harbors a mutation (P89S) that compromises enzymatic activity of the protein and genetically acts as a hypomorph. Transgenic overexpression of the Nme3 t-allele reduced t-haplotype transmission, proving it to be a distorter. We propose that the NME3 protein interacts with RHO signaling cascades to impair sperm motility through hyperactivation of SMOK, the wild-type form of the responder. This deleterious effect of the distorters is counter-balanced by the responder, SMOK(Tcr), a dominant-negative protein kinase exclusively expressed in t-sperm, thus permitting selfish behaviour and preferential transmission of the t-haplotype. In addition, the previously reported association of NME family members with RHO signaling in somatic cell motility and metastasis, in conjunction with our data involving RHO signaling in sperm motility, suggests a functional conservation between mechanisms for motility control in somatic cells and spermatozoa.

Lactic fermentation to improve the aroma of protein extracts of sweet lupin (Lupinus angustifolius).

Lupin protein extracts (LPE) are prone to the emission of a beany off-flavour during storage, which confines its application in foods. Fermentation of LPE using several lactic acid bacteria was conducted to reduce off-flavour formation in stored samples. The aroma profile of untreated LPE was compared to those of fermented protein extracts (LPEF). Hexanal and n-hexanol were used as indicator substances of progressing lipid oxidation. The most powerful odourants were evaluated by GC-olfactometry-flavour dilution analysis and identified according to their mass spectra, odour descriptions, and retention indices. Twenty two volatile substances with dilution factors equal to or higher than 100 were determined in both LPE and LPEF, amongst them n-pentanal, n-hexanal, 1-pyrroline, dimethyl trisulfide, 1-octen-3-one, 3-octen-2-one, 1-octen-3-ol, and ß-damascenone. The aroma profile was significantly modified by the fermentation process and the off-flavours were reduced and/or masked by newly formed compounds.

Author Correction: High-dimensional single-cell analysis predicts response to anti-PD-1 immunotherapy.

In the version of this article initially published, Figs. 5a,c and 6a were incorrect because of an error in a metadata spreadsheet that led to the healthy donor patient 2 (HD2) samples being used twice in the analysis of baseline samples and in the analysis at 12 weeks of anti-PD-1 therapy, while HD3 samples had not been used.

High-dimensional single-cell analysis predicts response to anti-PD-1 immunotherapy.

Immune-checkpoint blockade has revolutionized cancer therapy. In particular, inhibition of programmed cell death protein 1 (PD-1) has been found to be effective for the treatment of metastatic melanoma and other cancers. Despite a dramatic increase in progression-free survival, a large proportion of patients do not show durable responses. Therefore, predictive biomarkers of a clinical response are urgently needed. Here we used high-dimensional single-cell mass cytometry and a bioinformatics pipeline for the in-depth characterization of the immune cell subsets in the peripheral blood of patients with stage IV melanoma before and after 12 weeks of anti-PD-1 immunotherapy. During therapy, we observed a clear response to immunotherapy in the T cell compartment. However, before commencing therapy, a strong predictor of progression-free and overall survival in response to anti-PD-1 immunotherapy was the frequency of CD14+CD16-HLA-DRhi monocytes. We confirmed this by conventional flow cytometry in an independent, blinded validation cohort, and we propose that the frequency of monocytes in PBMCs may serve in clinical decision support.

The safety of anti PD-1 therapeutics for the treatment of melanoma.

INTRODUCTION: The introduction of immunotherapies into clinical practice has substantially improved the prognosis of metastatic melanoma patients as well as patients suffering from other cancers. The two FDA-approved checkpoint inhibitors against PD-1 (nivolumab and pembrolizumab) have been shown to significantly improve patient survival while being less toxic than previous treatment options. Areas covered: The current scientific literature on safety and adverse events (AEs) related to anti-PD-1 therapies has been investigated with special attention to case reports and to the latest results announced at the major clinical cancer and melanoma meetings, including ASCO (American Society of Clinical Oncology), ESMO (European Society of medical Oncology) and EADO (European Association of Dermato-Oncology) annual meetings. Expert opinion: Even though anti-PD-1 therapies are better tolerated than conventional chemo- or other immune-therapies, they still induce a plethora of AEs. Given the mechanism of action, it is supposed that most if not all of them are immune related. Fortunately, the majority are mild and manageable. However, due to the increase in patients' life expectancy, there is a substantial need to understand and prevent severe cutaneous, pulmonary, neurological and other AEs which have major impact on the quality of life. The safety profile after long term use of these medications is still unclear. In addition, non-steroid based immune interventions to control autoimmunity are still to be developed.

MEK inhibition and immune responses in advanced melanoma.

phase II and III clinical trials demonstrated modest anti- tumor activity of Binimetinib (MEK162) - a potent allosteric inhibitor of MEK1 and MEK2- in patients with advanced NRAS mutant melanoma. The analysis of the NEMO study in NRAS mutated melanoma, has shown that pre-treatment with immunotherapy improved the outcome of binimetinib therapy. We discuss this finding in the context of in vitro and in vivo effects of MEK inhibition on immuno-critical pathways and interactions.

Crystal structures of the viral protease Npro imply distinct roles for the catalytic water in catalysis.

Npro is a key effector protein of pestiviruses such as bovine viral diarrhea virus and abolishes host cell antiviral defense mechanisms. Synthesized as the N-terminal part of the viral polyprotein, Npro releases itself via an autoproteolytic cleavage, triggering its immunological functions. However, the mechanisms of its proteolytic action and its immune escape were unclear. Here, we present the crystal structures of Npro to 1.25 Å resolution. Structures of pre- and postcleavage intermediates identify three catalytically relevant elements. The trapping of the putative catalytic water reveals its distinct roles as a base, acid, and nucleophile. The presentation of the substrate further explains the enigmatic latency of the protease, ensuring a single in cis cleavage. Additionally, we identified a zinc-free, disulfide-linked conformation of the TRASH motif, an interaction hub of immune factors. The structure opens additional opportunities in utilizing Npro as an autocleaving fusion protein and as a pharmaceutical target.

Aroma development in high pressure treated beef and chicken meat compared to raw and heat treated.

Chicken breast and beef muscle were treated at 400 and 600 MPa for 15 min at 5 degrees C and compared to raw meat and a heated sample (100 degrees C for 15 min). Vacuum-packed beef meat with a smaller fraction of unsaturated fatty acids showed better oxidative stability during 14 days of cold storage, as shown by a low steady-state level of hydroperoxide values, than vacuum-packed chicken meat. Accordingly, the critical pressures of 400 MPa and 600 MPa for chicken breast and beef sirloin, respectively, were established. Volatiles released after opening of the meat bags or during storage of open meat bags, simulating consumer behaviour, were measured under conditions mimicking eating. Quantitative and olfactory analysis of pressurised meat gave a total of 46 flavour volatiles, mainly alcohols (11), aldehydes (15), and ketones (11), but all in low abundance after 14 days of storage. Overall, beef meat contained less volatiles and in lower abundance (factor of 5) compared to chicken meat. The most important odour active volatiles (GC-O) were well below the detection thresholds necessary to impart a perceivable off-flavour. Lipid oxidation was significantly accelerated during 24h of cold storage in both cooked chicken and beef when exposed to oxygen, while the pressurised and oxygen-exposed chicken and beef meat remained stable. Pressure treatment of beef and chicken did not induce severe changes of their raw aroma profiles.