Which Sit-to-Stand Test Best Differentiates Functional Capacity in Older People?

ABSTRACT: The sit-to-stand test provides insight into age-related functional capacity; however, there are various variants of sit-to-stand, and we do not know which of these better discriminates against age-related functional capacity. Our study aimed to compare the age-related functional capacity in older people by evaluating sit-to-stand power variants, using young individuals as a reference. A cross-sectional study was conducted in 102 adults (57 women) aged 60-80 and 105 adults (54 women) aged 20-30. Participants performed five times sit-to-stand (5 sit-to-stand), 30-second sit-to-stand, and 1-min sit-to-stand. Z scores were obtained for each sit-to-stand variant using power (W), relative (W/kg), and allometric (W/m 2 ) normalization methods. A mixed repeated-measures analysis of variance assessed the interactions among the sit-to-stand variants, normalization methods, sex, physical activity, and tobacco history. A significant interaction between sit-to-stand variants, normalization methods, and sex ( P = 0.002) was found. The mean effect of sit-to-stand variants revealed that the 1-min sit-to-stand had the lowest Z score ( P < 0.05). Significant variations were observed between sit-to-stand variants in all normalization methods for women ( P < 0.001). However, in men, only the difference between 5 sit-to-stand and 1-min sit-to-stand remained consistent across normalization methods ( P < 0.05). Our findings highlight the efficacy of 1-min sit-to-stand in distinguishing age-related functional capacity over the other sit-to-stand tests, especially in women.

Enhancing the efficiency of magnetically driven carbon nitride-based nanocomposites with magnetic nanoflowers for the removal of methylene blue dye at neutral pH.

The present study focuses on the elaboration of magnetic nanocomposites by the in situ incorporation of magnetite (Fe3O4) nanoparticles (NPs) with spherical and nanoflower-like morphologies in graphitic carbon nitride (g-C3N4) sheets using two different synthetic routes. Nanomaterials are characterized by TEM, SEM, XRD, FTIR, BET, zetametry, vibrating sample magnetometry, and UV-vis absorption spectroscopy. The decoration of the carbon nitride matrix with the magnetic NPs enhanced optical and textural properties. The influence of the morphology of the magnetic NPs on the adsorptive and photocatalytic properties of the nanocomposites under different pH conditions (4.5, 6.9, and 10.6) was assessed from batch tests to remove methylene blue (MB) from aqueous solutions. In extreme pH conditions, the nanocomposites exhibited lower or equivalent MB removal capacity compared to the pure g-C3N4. However, at neutral medium, the nanocomposite with incorporated Fe3O4 nanoflowers showed a significantly higher removal efficiency (80.7%) due to the combination of a high adsorption capacity and a good photocatalytic activity in this pH region. The proposed nanocomposite is a promising alternative to remove cationic dyes from water by magnetic assistance, since no pH adjustment of the polluted effluent is required, reducing costs and environmental impact in the dyeing industry.

Inferring secretory and metabolic pathway activity from omic data with secCellFie.

Understanding protein secretion has considerable importance in biotechnology and important implications in a broad range of normal and pathological conditions including development, immunology, and tissue function. While great progress has been made in studying individual proteins in the secretory pathway, measuring and quantifying mechanistic changes in the pathway's activity remains challenging due to the complexity of the biomolecular systems involved. Systems biology has begun to address this issue with the development of algorithmic tools for analyzing biological pathways; however most of these tools remain accessible only to experts in systems biology with extensive computational experience. Here, we expand upon the user-friendly CellFie tool which quantifies metabolic activity from omic data to include secretory pathway functions, allowing any scientist to infer properties of protein secretion from omic data. We demonstrate how the secretory expansion of CellFie (secCellFie) can help predict metabolic and secretory functions across diverse immune cells, hepatokine secretion in a cell model of NAFLD, and antibody production in Chinese Hamster Ovary cells.

Two national mammography quality certification programs in Brazil: Framework and main outcomes between 2017 and 2021.

BACKGROUND: This study aims to provide an academic medical overview of the framework and key outcomes of two mammography quality certification programs in Brazil. METHODS: These programs assess radiation dose and phantom image quality in mammography units through a postal system. Each unit that passes this initial assessment is required to submit a sample of copies of five complete examinations. The quality of the patient images and reports is then reviewed by radiologists and medical physicist experts. Additionally, the number of mammography units and mammography coverage in the target population, were assessed. RESULTS: During the study period, 1007 units applied to the certification programs, and 934 (92.8%) successfully passed the assessment of radiation dose and phantom image quality. Out of these, 556 (59.5%) also passed the review of clinical image quality and reports, earning certification. The main issues related to mammogram and report quality were associated with the performance of radiographers (in terms of positioning) and radiologists (in terms of interpretation). On average, there are more than two mammography units/10,000 women in the target group. The screening mammography coverage in this group is 26.3% for women relying exclusively on the public healthcare and 58.1% for women with private healthcare plans. CONCLUSION: This study demonstrates the suitability of the framework adopted by national mammography quality certification programs in a middle-income country. These programs are carried out by relatively small workforce and at reasonable costs, utilizing postal resources to cover the large number of existing mammographic units and the vast distances within the country. POLICY STATEMENT: All mammography services in Brazil must adhere to the quality requirements for examinations and reference values for radiation dose to women established by the Ministry of Health. This ensures standardized conditions for early detection of breast cancer and minimizes the risk associated with x-rays.

Inferring secretory and metabolic pathway activity from omic data with secCellFie.

Understanding protein secretion has considerable importance in the biotechnology industry and important implications in a broad range of normal and pathological conditions including development, immunology, and tissue function. While great progress has been made in studying individual proteins in the secretory pathway, measuring and quantifying mechanistic changes in the pathway's activity remains challenging due to the complexity of the biomolecular systems involved. Systems biology has begun to address this issue with the development of algorithmic tools for analyzing biological pathways; however most of these tools remain accessible only to experts in systems biology with extensive computational experience. Here, we expand upon the user-friendly CellFie tool which quantifies metabolic activity from omic data to include secretory pathway functions, allowing any scientist to infer protein secretion capabilities from omic data. We demonstrate how the secretory expansion of CellFie (secCellFie) can be used to predict metabolic and secretory functions across diverse immune cells, hepatokine secretion in a cell model of NAFLD, and antibody production in Chinese Hamster Ovary cells.

L-Lysine-Coated Magnetic Core-Shell Nanoparticles for the Removal of Acetylsalicylic Acid from Aqueous Solutions.

Nanotechnologies based on magnetic materials have been successfully used as efficient and reusable strategies to remove pharmaceutical residuals from water. This paper focuses on the fabrication, characterization, and application of ferrite-based magnetic nanoparticles functionalized with L-lysine as potential nanoadsorbents to remove acetylsalicylic acid (ASA) from water. The proposed nanomaterials are composed of highly magnetic and chemically stable core-shell nanoparticles covered with an adsorptive layer of L-lysine (CoFe2O4-γ-Fe2O3-Lys). The nanoadsorbents were elaborated using the coprecipitation method in an alkaline medium, leading to nanoparticles with two different mean sizes (13.5 nm and 8.5 nm). The samples were characterized by XRD, TEM, FTIR, XPS, Zetametry, BET, and SQUID magnetometry. The influence of time, pH, and pollutant concentration was evaluated from batch studies using 1.33 g/L of the nanoadsorbents. The Freundlich isotherm best adjusted the adsorption data. The adsorption process exhibited a pseudo-second-order kinetic behavior. The optimal pH for adsorption was around 4-6, with a maximum adsorption capacity of 16.4 mg/g after 150 min of contact time. Regeneration tests also showed that the proposed nanomaterials are reusable. The set of results proved that the nanoadsorbents can be potentially used to remove ASA from water and provide relevant information for their application in large-scale designs.

Trem2 deletion enhances tau dispersion and pathology through microglia exosomes.

BACKGROUND: Alzheimer's disease (AD) is a neurodegenerative disorder that manifests sequential Aß and tau brain pathology with age-dependent onset. Variants in the microglial immune receptor TREM2 are associated with enhanced risk of onset in sporadic Alzheimer's disease (AD). While recent studies suggest TREM2 dysfunction can aggravate tau pathology, mechanisms underlying TREM2-dependent modulation of tau pathology remains elusive. METHODS: Here, we characterized differences in progressive tau spreading from the medial entorhinal cortex (MEC) to the hippocampus in wildtype (WT) and Trem2 knockout (KO) mice by injection of AAV-P301L tau into the MEC, and correlated changes in hippocampal tau histopathology with spatial and fear memory. We also compared effects of intraneuronal dispersion between cultured microglia and neurons using a microfluidic dispersion assay, analyzed differences in microglial tau trafficking following uptake, and quantified exosomal tau secretion and pathogenicity from purified WT and Trem2 KO exosomes. RESULTS: Trem2 deletion in mice (Trem2 KO) can enhance tau spreading from the medial entorhinal cortex (MEC) to the hippocampus, which coincides with impaired synaptic function and memory behavior. Trem2 deletion in microglia enhances intraneuronal dispersion of tau in vitro between neuronal layers cultured in a microfluidic chamber, and the presence of exosome inhibitors can significantly reduce tau in exosomes and extracellular media from tau-loaded microglia. Although microglial Trem2 deletion has no effect on tau uptake, Trem2 deletion enhances distribution to endosomal and cellular pre-exosomal compartments following internalization. Trem2 deletion has little effect on exosome size, however, proteomic analysis indicates that Trem2 deletion can modulate changes in the microglial proteomic landscape with tau and LPS/ATP treatment conditions associated with exosome induction. Furthermore, exosomes from Trem2 KO microglia show elevated tau levels, and feature enhanced tau-seeding capacity in a tau FRET reporter line compared to exosomes from WT microglia. CONCLUSION: Together, our results reveal a role for Trem2 in suppressing exosomal tau pathogenicity, and demonstrates that Trem2 deletion can enhance tau trafficking, distribution and seeding through microglial exosomes.

A numerical study on the interplay between the intra-particle and interparticle characteristics in bimagnetic soft/soft and hard/soft ultrasmall nanoparticle assemblies.

A mesoscopic scale approach and the Monte Carlo (MC) method have been employed to study the exchange bias behaviour of MnFe2O4 (soft)/maghemite (soft) and CoFe2O4 (hard)/maghemite (soft) nanoparticles (NPs) of size ∼ 3 nm in dense and diluted assemblies at low temperatures. The analysis of our MC results clearly shows that in the powder samples the contribution to the exchange bias field (H ex) and the coercivity (H c) comes mainly from the intraparticle core/shell structure in the hard/soft sample and that the interplay between the internal characteristics and the interparticle interactions is more important in the soft/soft samples where the dipolar strength is enhanced. In the diluted frozen ferrofluid samples where interparticle exchange interactions are absent and the role of the dipolar interactions is not significant the exchange bias effects are reduced, and they come from the intra particle structure. The variation of H ex and H c with the applied cooling field well reproduces the experimental findings and sheds light on the key mechanisms of the observed magnetic behaviour. Our results demonstrate the possibility to control the magnetic behaviour of nanostructures by using properly chosen core/shell bimagnetic nanoparticles.

Hybrid magnetic CoFe2O4@γ-Fe2O3@CTAB nanocomposites as efficient and reusable adsorbents for Remazol Brilliant Blue R dye.

The main goal of the present survey was to elaborate, characterize and evaluate the efficiency of ferrite-based nanoparticles modified with cetyltrimethylammonium bromide (CTAB) as potential magnetic nanoadsorbents to remove Remazol Brilliant Blue R (RBBR) from water. It is proposed an innovative nanomaterial architecture based on highly magnetic and chemically stable core@shell nanoparticles covered by an adsorptive surface layer of CTAB (CoFe2O4@γ-Fe2O3@CTAB). Samples of two different mean sizes (7.5 and 14.6 nm) were synthesized using a hydrothermal coprecipitation followed by surface treatment and functionalization. Batch tests were performed to evaluate the influence of contact time, temperature, pH, shaking rate, presence of interferents and mean size on the performance of the proposed nanomaterials. The kinetics of the adsorption process followed the pseudo-second-order model with an equilibrium time of 20 min. The adsorption capacity was estimated by the Langmuir isotherm model and was found to be 56.3 mg/g (smaller size) and 45.6 mg/g (larger size) at pH = 3 and a shaking rate of 400 rpm. The process was spontaneous, exothermic, and showed increased randomness. Sulphate ions negatively impacted the removal of RBBR. The best performance of the nanoadsorbent based on smaller mean sizes can be correlated to its larger surface area. Regeneration and readsorption tests showed that the nanoadsorbents retain more than 80% of their original removal capacity, therefore they can be effectively recycled and reused.

Combined Inhibition of AKT and KIT Restores Expression of Programmed Cell Death 4 (PDCD4) in Gastrointestinal Stromal Tumor.

The majority of gastrointestinal stromal tumor (GIST) patients develop resistance to the first-line KIT inhibitor, imatinib mesylate (IM), through acquisition of secondary mutations in KIT or bypass signaling pathway activation. In addition to KIT, AKT is a relevant target for inhibition, since the PI3K/AKT pathway is crucial for IM-resistant GIST survival. We evaluated the activity of a novel pan-AKT inhibitor, MK-4440 (formerly ARQ 751), as monotherapy and in combination with IM in GIST cell lines and preclinical models with varying IM sensitivities. Dual inhibition of KIT and AKT demonstrated synergistic effects in IM-sensitive and -resistant GIST cell lines. Proteomic analyses revealed upregulation of the tumor suppressor, PDCD4, in combination treated cells. Enhanced PDCD4 expression correlated to increased cell death. In vivo studies revealed superior efficacy of MK-4440/IM combination in an IM-sensitive preclinical model of GIST compared with either single agent. The combination demonstrated limited efficacy in two IM-resistant models, including a GIST patient-derived xenograft model possessing an exon 9 KIT mutation. These studies provide strong rationale for further use of AKT inhibition in combination with IM in primary GIST; however, alternative agents will need to be tested in combination with AKT inhibition in the resistant setting.

PTEN deficiency leads to proteasome addiction: a novel vulnerability in glioblastoma.

BACKGROUND: Glioblastoma (GBM) is the most common primary brain tumor in adults with a median survival of approximately 15 months; therefore, more effective treatment options for GBM are required. To identify new drugs targeting GBMs, we performed a high-throughput drug screen using patient-derived neurospheres cultured to preferentially retain their glioblastoma stem cell (GSC) phenotype. METHODS: High-throughput drug screening was performed on GSCs followed by a dose-response assay of the 5 identified original "hits." A PI3K/mTOR dependency to a proteasome inhibitor (carfilzomib), was confirmed by genetic and pharmacologic experiments. Proteasome Inhibition Response Signatures were derived from proteomic and bioinformatic analysis. Molecular mechanism of action was determined using three-dimensional (3D) GBM-organoids and preclinical orthotopic models. RESULTS: We found that GSCs were highly sensitive to proteasome inhibition due to an underlying dependency on an increased protein synthesis rate, and loss of autophagy, associated with PTEN loss and activation of the PI3K/mTOR pathway. In contrast, combinatory inhibition of autophagy and the proteasome resulted in enhanced cytotoxicity specifically in GSCs that did express PTEN. Finally, proteasome inhibition specifically increased cell death markers in 3D GBM-organoids, suppressed tumor growth, and increased survival of mice orthotopically engrafted with GSCs. As perturbations of the PI3K/mTOR pathway occur in nearly 50% of GBMs, these findings suggest that a significant fraction of these tumors could be vulnerable to proteasome inhibition. CONCLUSIONS: Proteasome inhibition is a potential synthetic lethal therapeutic strategy for GBM with proteasome addiction due to a high protein synthesis rate and autophagy deficiency.

Soluble SORLA Enhances Neurite Outgrowth and Regeneration through Activation of the EGF Receptor/ERK Signaling Axis.

SORLA is a transmembrane trafficking protein associated with Alzheimer's disease risk. Although SORLA is abundantly expressed in neurons, physiological roles for SORLA remain unclear. Here, we show that cultured transgenic neurons overexpressing SORLA feature longer neurites, and accelerated neurite regeneration with wounding. Enhanced release of a soluble form of SORLA (sSORLA) is observed in transgenic mouse neurons overexpressing human SORLA, while purified sSORLA promotes neurite extension and regeneration. Phosphoproteomic analyses demonstrate enrichment of phosphoproteins related to the epidermal growth factor (EGFR)/ERK pathway in SORLA transgenic mouse hippocampus from both genders. sSORLA coprecipitates with EGFR in vitro, and sSORLA treatment increases EGFR Y1173 phosphorylation, which is involved in ERK activation in cultured neurons. Furthermore, sSORLA triggers ERK activation, whereas pharmacological EGFR or ERK inhibition reverses sSORLA-dependent enhancement of neurite outgrowth. In search for downstream ERK effectors activated by sSORLA, we identified upregulation of Fos expression in hippocampus from male mice overexpressing SORLA by RNAseq analysis. We also found that Fos is upregulated and translocates to the nucleus in an ERK-dependent manner in neurons treated with sSORLA. Together, these results demonstrate that sSORLA is an EGFR-interacting protein that activates EGFR/ERK/Fos signaling to enhance neurite outgrowth and regeneration.SIGNIFICANCE STATEMENT SORLA is a transmembrane trafficking protein previously known to reduce the levels of amyloid-ß, which is critical in the pathogenesis of Alzheimer's disease. In addition, SORLA mutations are a risk factor for Alzheimer's disease. Interestingly, the SORLA ectodomain is cleaved into a soluble form, sSORLA, which has been shown to regulate cytoskeletal signaling pathways and cell motility in cells outside the nervous system. We show here that sSORLA binds and activates the EGF receptor to induce downstream signaling through the ERK serine/threonine kinase and the Fos transcription factor, thereby enhancing neurite outgrowth. These findings reveal a novel role for sSORLA in promoting neurite regeneration through the EGF receptor/ERK/Fos pathway, thereby demonstrating a potential neuroprotective mechanism involving SORLA.

PKCλ/ι Loss Induces Autophagy, Oxidative Phosphorylation, and NRF2 to Promote Liver Cancer Progression.

Oxidative stress plays a critical role in liver tissue damage and in hepatocellular carcinoma (HCC) initiation and progression. However, the mechanisms that regulate autophagy and metabolic reprogramming during reactive oxygen species (ROS) generation, and how ROS promote tumorigenesis, still need to be fully understood. We show that protein kinase C (PKC) λ/ι loss in hepatocytes promotes autophagy and oxidative phosphorylation. This results in ROS generation, which through NRF2 drives HCC through cell-autonomous and non-autonomous mechanisms. Although PKCλ/ι promotes tumorigenesis in oncogene-driven cancer models, emerging evidence demonstrate that it is a tumor suppressor in more complex carcinogenic processes. Consistently, PKCλ/ι levels negatively correlate with HCC histological tumor grade, establishing this kinase as a tumor suppressor in liver cancer.

Core-Shell Bimagnetic Nanoadsorbents for Hexavalent Chromium Removal from Aqueous Solutions.

Novel nanoadsorbents based on core-shell bimagnetic nanoparticles (CoFe2O4@É£-Fe2O3) with two different mean sizes were elaborated, characterized and applied as potential sorbents for Cr(VI) removal from aqueous solutions through magnetically assisted chemical separation. The nanoadsorbents were characterized by XRD, TEM, FTIR, XPS, potentiometric-conductometric titrations, BET and vibrating sample magnetometry. The influence of contact time, shaking rate, pH, pollutant concentration, temperature and competing ions on Cr(VI) adsorption were evaluated. The results were analyzed in the framework of Langmuir and Freundlich models to evaluate the maximum adsorption capacity and the extent of affinity. The nanoadsorbents showed a good selectivity for Cr(VI) adsorption and were more effective at pH = 2.5, with a shaking rate of 400 RPM. The adsorption process was spontaneous, endothermic and presented an increased randomness. The contact time required to reach the equilibrium was relatively short and the kinetic date followed the pseudo-second-order model. The maximum adsorption capacity was nearly 40% higher for the nanoadsorbent of smaller mean size due to its higher surface area. Regeneration studies revealed that the nanoadsorbents can be recovered for reuse. These results indicate that prepared nanoadsorbents can be used as a powerful tool for Cr(VI) removal from contaminated water.

The Secretion of miR-200s by a PKCζ/ADAR2 Signaling Axis Promotes Liver Metastasis in Colorectal Cancer.

Most colorectal cancer (CRC)-related deaths are due to liver metastases. PKCζ is a tumor suppressor in CRC with reduced expression in metastasis. Given the importance of microRNAs (miRNAs) in regulating cellular plasticity, we performed an unbiased screening and identified the miR-200 family as the most relevant miRNAs downregulated by PKCζ deficiency. The regulation of the intracellular levels of miR-200 by PKCζ is post-transcriptional and involves their secretion in extracellular vesicles. Here, we identified ADAR2 as a direct substrate of PKCζ in CRC cells. Phosphorylation of ADAR2 regulates its editing activity, which is required to maintain miR-200 steady-state levels, suggesting that the PKCζ/ADAR2 axis regulates miR-200 secretion through RNA editing. Loss of this axis results in epithelial-to-mesenchymal transition (EMT) and increased liver metastases, which can be inhibited in vivo by blocking miR-200 release. Therefore, the PKCζ/ADAR2 axis is a critical regulator of CRC metastases through modulation of miR-200 levels.

Targeted proteomics in urinary extracellular vesicles identifies biomarkers for diagnosis and prognosis of prostate cancer.

Rapid and reliable diagnosis of prostate cancer (PCa) is highly desirable as current used methods lack specificity. In addition, identification of PCa biomarkers that can classify patients into high- and low-risk groups for disease progression at early stage will improve treatment decision-making. Here, we describe a set of protein-combination panels in urinary extracellular vesicles (EVs), defined by targeted proteomics and immunoblotting techniques that improve early non-invasive detection and stratification of PCa patients.We report a two-protein combination in urinary EVs that classifies benign and PCa patients (ADSV-TGM4), and a combination of five proteins able to significantly distinguish between high- and low-grade PCa patients (CD63-GLPK5-SPHM-PSA-PAPP). Proteins composing the panels were validated by immunohistochemistry assays in tissue microarrays (TMAs) confirming a strong link between the urinary EVs proteome and alterations in PCa tissues. Moreover, ADSV and TGM4 abundance yielded a high diagnostic potential in tissue and promising TGM4 prognostic power. These results suggest that the proteins identified in urinary EVs distinguishing high- and low grade PCa are a reflection of histological changes that may be a consequence of their functional involvement in PCa development. In conclusion, our study resulted in the identification of protein-combination panels present in urinary EVs that exhibit high sensitivity and specificity for PCa detection and patient stratification. Moreover, our study highlights the potential of targeted proteomic approaches-such as selected reaction monitoring (SRM)-as diagnostic assay for liquid biopsies via urinary EVs to improve diagnosis and prognosis of suspected PCa patients.

Control of Paneth Cell Fate, Intestinal Inflammation, and Tumorigenesis by PKCλ/ι.

Paneth cells are a highly specialized population of intestinal epithelial cells located in the crypt adjacent to Lgr5(+) stem cells, from which they differentiate through a process that requires downregulation of the Notch pathway. Their ability to store and release antimicrobial peptides protects the host from intestinal pathogens and controls intestinal inflammation. Here, we show that PKCλ/ι is required for Paneth cell differentiation at the level of Atoh1 and Gfi1, through the control of EZH2 stability by direct phosphorylation. The selective inactivation of PKCλ/ι in epithelial cells results in the loss of mature Paneth cells, increased apoptosis and inflammation, and enhanced tumorigenesis. Importantly, PKCλ/ι expression in human Paneth cells decreases with progression of Crohn's disease. Kaplan-Meier survival analysis of colorectal cancer (CRC) patients revealed that low PRKCI levels correlated with significantly worse patient survival rates. Therefore, PKCλ/ι is a negative regulator of intestinal inflammation and cancer through its role in Paneth cell homeostasis.

Amino Acid Activation of mTORC1 by a PB1-Domain-Driven Kinase Complex Cascade.

The mTORC1 complex is central to the cellular response to changes in nutrient availability. The signaling adaptor p62 contributes to mTORC1 activation in response to amino acids and interacts with TRAF6, which is required for the translocation of mTORC1 to the lysosome and the subsequent K63 polyubiquitination and activation of mTOR. However, the signal initiating these p62-driven processes was previously unknown. Here, we show that p62 is phosphorylated via a cascade that includes MEK3/6 and p38δ and is driven by the PB1-containing kinase MEKK3. This phosphorylation results in the recruitment of TRAF6 to p62, the ubiquitination and activation of mTOR, and the regulation of autophagy and cell proliferation. Genetic inactivation of MEKK3 or p38δ mimics that of p62 in that it leads to inhibited growth of PTEN-deficient prostate organoids. Analysis of human prostate cancer samples showed upregulation of these three components of the pathway, which correlated with enhanced mTORC1 activation.

Multicenter experiment for quality control of peptide-centric LC-MS/MS analysis - A longitudinal performance assessment with nLC coupled to orbitrap MS analyzers.

Proteomic technologies based on mass spectrometry (MS) have greatly evolved in the past years, and nowadays it is possible to routinely identify thousands of peptides from complex biological samples in a single LC-MS/MS experiment. Despite the advancements in proteomic technologies, the scientific community still faces important challenges in terms of depth and reproducibility of proteomics analyses. Here, we present a multicenter study designed to evaluate long-term performance of LC-MS/MS platforms within the Spanish Proteomics Facilities Network (ProteoRed-ISCIII). The study was performed under well-established standard operating procedures, and demonstrated that it is possible to attain qualitative and quantitative reproducibility over time. Our study highlights the importance of deploying quality assessment metrics routinely in individual laboratories and in multi-laboratory studies. The mass spectrometry data have been deposited to the ProteomeXchange Consortium with the data set identifier PXD000205.This article is part of a Special Issue entitled: HUPO 2014.

Repression of Intestinal Stem Cell Function and Tumorigenesis through Direct Phosphorylation of ß-Catenin and Yap by PKCζ.

Intestinal epithelial homeostasis requires continuous renewal supported by stem cells located in the base of the crypt. Disruption of this balance results in failure to regenerate and initiates tumorigenesis. The ß-catenin and Yap pathways in Lgr5+ stem cells have been shown to be central to this process. However, the precise mechanisms by which these signaling molecules are regulated in the stem cell population are not totally understood. Protein kinase C ζ (PKCζ) has been previously demonstrated to be a negative regulator of intestinal tumorigenesis. Here, we show that PKCζ suppresses intestinal stem cell function by promoting the downregulation of ß-catenin and Yap through direct phosphorylation. PKCζ deficiency results in increased stem cell activity in organoid cultures and in vivo, accounting for the increased tumorigenic and regenerative activity response of Lgr5+-specific PKCζ-deficient mice. This demonstrates that PKCζ is central to the control of stem cells in intestinal cancer and homeostasis.