Serial single-cell profiling analysis of metastatic TNBC during Nab-paclitaxel and pembrolizumab treatment

Purpose: Immunotherapy has recently been shown to improve outcomes for advanced PD-L1-positive triple-negative breast cancer (TNBC) in the Impassion130 trial, leading to FDA approval of the first immune checkpoint inhibitor in combination with taxane chemotherapy. To further develop predictive biomarkers and improve therapeutic efficacy of the combination, interrogation of the tumor immune microenvironment before therapy as well as during each component of treatment is crucial. Here we use single-cell RNA sequencing (scRNA-seq) on tumor biopsies to assess immune cell changes from two patients with advanced TNBC treated in a prospective trial at predefined serial time points, before treatment, on taxane chemotherapy and on chemo-immunotherapy. Methods: Both patients (one responder and one progressor) received the trial therapy, in cycle 1 nab-paclitaxel given as single agent, in cycle 2 nab-paclitaxel in combination with pembrolizumab. Tumor core biopsies were obtained at baseline, 3 weeks (after cycle 1, chemotherapy alone) and 6 weeks (after cycle 2, chemo-immunotherapy). Single-cell RNA sequencing (scRNA-seq) of both cancer cells and infiltrating immune cells isolated were performed from fresh tumor core biopsy specimens by 10 × chromium sequencing. Results: ScRNA-seq analysis showed significant baseline heterogeneity of tumor-infiltrating immune cell populations between the two patients as well as modulation of the tumor microenvironment by chemotherapy and immunotherapy. In the responding patient there was a population of PD-1high-expressing T cells which significantly decreased after nab-paclitaxel plus pembrolizumab treatment as well as a presence of tissue-resident memory T cells (TRM). In contrast, tumors from the patient with rapid disease progression showed a prevalent and persistent myeloid compartment. Conclusions: Our study provides a deep cellular analysis of on-treatment changes during chemo-immunotherapy for advanced TNBC, demonstrating not only feasibility of single-cell analyses on serial tumor biopsies but also the heterogeneity of TNBC and differences in on-treatment changes in responder versus progressor

Bioaccessibility of phenolic compounds and antioxidant activity in raw and pickled capers

Capers are excellent sources of phenolic compounds, which possess antioxidant activity and a variety of health benefits. Capers are often not eaten fresh due to their bitterness, instead, they are usually brined and eaten as pickles. For phenolics to display beneficial effects, they should be bioaccessible. This research aimed to investigate the bioaccessibility values of phenolics in raw and pickled capers. Before and after in vitro digestion, total phenolic content (TPC), total flavonoid content (TFC), ABTS, DPPH, and FRAP antioxidant activities and individual phenolics were determined in raw (RC) and pickled capers (PC). TPC, TFC, and antioxidant activity of capers were not affected by pickling. The bioaccessibility of TPC in RC and PC was 77.8% and 72.9%, respectively. In vitro digestion decreased ABTS, DPPH, and FRAP antioxidant activity values in RC and PC significantly (p < 0.05). Rutin (quercetin-3-O-rutinoside) and kaempferol-3-O-rutinoside were found to be the main phenolic compounds before and after in vitro digestion in all caper samples. After in vitro digestion, rutin content decreased, while kaempferol-3-O-rutinoside content in capers was stable. The results suggest that capers are really good sources of bioaccessible phenolics

Bioavailability of quercetin

Quercetin is generally present as quercetin glycoside in nature and involves quercetin aglycone conjugated to sugar moieties such as glucose or rutinose. Quercetin has been reported to exhibit antioxidative, anti-carcinogenic, anti-inflammatory, anti-aggregatory and vasodilating effects. Unfortunately, quercetin bioavailability is generally poor and several factors affect its bioavailability. Quercetin bioavailability varies widely between individuals. Gender may affect quercetin bioavailability, but there is no clear evidence. There has been little research looking for the effects of age and vitamin C status on bioavailability of quercetin supplements, but there is no research seeking out the effects of age and vitamin C status on bioavailability of food-derived quercetin. Presence of sugar moieties increases bioavailability and differences in quercetin-conjugated glycosides affect bioavailability. For instance, onion-derived quercetin, which is mainly quercetin glucoside, is more bioavailable than apple-derived quercetin, which contains quercetin rhamnoside and quercetin galactoside. Quercetin is lipophilic compound, thus dietary fat enhances its bioavailability. Nondigestible fiber may also improve quercetin bioavailability. Quercetin bioavailability is greater when it is consumed as an integral food component. This study reviews and discusses factors affecting quercetin bioavailability. © 2016, Enviro Research Publishers. All rights reserved

Heavy metal levels in leafy vegetables and some selected fruits

The aim of this study was to determine the levels of Cu, Zn, Fe, Pb and Cd in various fruits (tomato, cherry, grape, strawberry) and vegetables (parsley, onion, lettuce, garlic, nettle, peppermint, rocket, spinach, dill, broad bean, chard, purslane, grapevine leaves) grown in Manisa region. Flame and Graphite Furnace Atomic absorption spectrometry was used to estimate and evaluate the levels of these metals. Detected levels ranged from 0.56 to 329.7, 0.01 to 5.67, 0.26 to 30.68, 0.001 to 0.97 and 0 to 0.06 mu g/g for Fe, Cu, Zn, Pb and Cd, respectively. While the highest mean levels of Cu and Zn were detected in grapevine leaves, the lowest mean levels of Fe and Pb were detected in nettle. Cd was not detected in most of the fruits and vegetables studied. The estimated daily intakes of Cu, Zn, Fe, Pb and Cd through fruits and vegetables were found to be below the maximum tolerable levels recommended by FAO/WHO. The element concentrations of fruits and vegetables analyzed in this study were within safety baseline levels for human consumption

Effects of deboning methods on chemical composition and some properties of beef and turkey meat

The objective of this research was to compare some chemical and physical characteristics of mechanically deboned and hand deboned turkey meat and beef. Samples were analyzed for proximate composition, cholesterol, TBA, calcium and iron contents, hunter colour parameters and fatty acid composition. Deboning method affected chemical composition of beef and turkey meat. Mechanical deboning resulted higher cholesterol values and calcium and iron content. In hand deboned turkey meat the most abundant fatty acids were C16:0, C18:1 and C18:2. Mechanical deboning process was increased the percent of C18:1, C18:2 and C18:3 in turkey meat. C16:0, C18:0 and C18:1 were the major fatty acids in mechanically deboned and hand deboned beef

Heavy metal levels in leafy vegetables and some selected fruits

The aim of this study was to determine the levels of Cu, Zn, Fe, Pb and Cd in various fruits (tomato, cherry, grape, strawberry) and vegetables (parsley, onion, lettuce, garlic, nettle, peppermint, rocket, spinach, dill, broad bean, chard, purslane, grapevine leaves) grown in Manisa region. Flame and Graphite Furnace Atomic absorption spectrometry was used to estimate and evaluate the levels of these metals. Detected levels ranged from 0.56 to 329.7, 0.01 to 5.67, 0.26 to 30.68, 0.001 to 0.97 and 0 to 0.06 μg/g for Fe, Cu, Zn, Pb and Cd, respectively. While the highest mean levels of Cu and Zn were detected in grapevine leaves, the lowest mean levels of Fe and Pb were detected in nettle. Cd was not detected in most of the fruits and vegetables studied. The estimated daily intakes of Cu, Zn, Fe, Pb and Cd through fruits and vegetables were found to be below the maximum tolerable levels recommended by FAO/WHO. The element concentrations of fruits and vegetables analyzed in this study were within safety baseline levels for human consumption. © 2010 Springer Basel AG