The ability of CD8
T lymphocytes to eliminate tumors is limited by their ability to engender an immunosuppressive microenvironment. Here we describe a subset of tumor-infiltrating CD8
T cells marked ...by high expression of the immunosuppressive ATP ecto-nucleotidase CD39. The frequency of CD39
CD8
T cells increased with tumor growth but was absent in lymphoid organs. Tumor-infiltrating CD8
T cells with high CD39 expression exhibited features of exhaustion, such as reduced production of TNF and IL2 and expression of coinhibitory receptors. Exhausted CD39
CD8
T cells from mice hydrolyzed extracellular ATP, confirming that CD39 is enzymatically active. Furthermore, exhausted CD39
CD8
T cells inhibited IFNγ production by responder CD8
T cells. In specimens from breast cancer and melanoma patients, CD39
CD8
T cells were present within tumors and invaded or metastatic lymph nodes, but were barely detectable within noninvaded lymph nodes and absent in peripheral blood. These cells exhibited an exhausted phenotype with impaired production of IFNγ, TNF, IL2, and high expression of coinhibitory receptors. Although T-cell receptor engagement was sufficient to induce CD39 on human CD8
T cells, exposure to IL6 and IL27 promoted CD39 expression on stimulated CD8
T cells from human or murine sources. Our findings show how the tumor microenvironment drives the acquisition of CD39 as an immune regulatory molecule on CD8
T cells, with implications for defining a biomarker of T-cell dysfunction and a target for immunotherapeutic intervention.
The tumor microenvironment elicits a subset of functionally exhausted CD8
T cells by creating conditions that induce cell surface expression of CD39, an immunosuppressive molecule that can be therapeutically targeted to restore effector T-cell function.
.
Pancreatic ductal adenocarcinoma (PDAC) is usually detected late in the disease process. Clinical workup through imaging and tissue biopsies is often complex and expensive due to a paucity of ...reliable biomarkers. We used an advanced multiplexed plasmonic assay to analyze circulating tumor-derived extracellular vesicles (tEVs) in more than 100 clinical populations. Using EV-based protein marker profiling, we identified a signature of five markers (PDAC
signature) for PDAC detection. In our prospective cohort, the accuracy for the PDAC
signature was 84% 95% confidence interval (CI), 69 to 93% but only 63 to 72% for single-marker screening. One of the best markers, GPC1 alone, had a sensitivity of 82% (CI, 60 to 95%) and a specificity of 52% (CI, 30 to 74%), whereas the PDAC
signature showed a sensitivity of 86% (CI, 65 to 97%) and a specificity of 81% (CI, 58 to 95%). The PDAC
signature of tEVs offered higher sensitivity, specificity, and accuracy than the existing serum marker (CA 19-9) or single-tEV marker analyses. This approach should improve the diagnosis of pancreatic cancer.
•The influence of key variables in a Sn cathode filter-press cell was studied.•The performance of Sn plate was compared with previous results using Pb.•As with Pb, the influence of current density ...using Sn was especially remarkable.•Results with Sn plate cathodes outperformed those with Pb plate.•These results reinforce the interest in Sn for CO2 electrovalorisation to formate.
Electrochemical reduction has been pointed out as a promising method for CO2 valorisation into useful chemicals. This paper studies the influence of key variables on the performance of an experimental system for continuous electro-reduction of CO2 to formate, when a tin plate is used as working electrode. Particular emphasis is placed on comparing the performance of Sn and Pb as cathodes. As was previously found with Pb, the influence of current density (“j”) using Sn was particularly noteworthy, and when j was raised up to a limit value of 8.5mAcm−2, important increases of the rate of formate production were observed at the expense of lowering the Faradaic efficiency. However, unlike what was found with Pb, the performance using Sn improved when the electrolyte flow rate/electrode area ratio was increased within the range studied (0.57–2.3mLmin−1cm−2). In this way, the use of Sn as cathode allowed achieving rates of formate production that were 25% higher than the maximum rates obtained with Pb, together with Faradaic efficiencies close to 70%, which were 15 points higher than those with Pb. These results reinforce the interest in Sn as electrode material in the electro-reduction of CO2 to formate.
Organoids are becoming increasingly relevant in biology and medicine for their physiological complexity and accuracy in modeling human disease. To fully assess their biological profile while ...preserving their spatial information, spatiotemporal imaging tools are warranted. While previously developed imaging techniques, such as four‐dimensional (4D) live imaging and light‐sheet imaging have yielded important clinical insights, these technologies lack the combination of cyclic and multiplexed analysis. To address these challenges, bioorthogonal click chemistry is applied to display the first demonstration of multiplexed cyclic imaging of live and fixed patient‐derived glioblastoma tumor organoids. This technology exploits bioorthogonal click chemistry to quench fluorescent signals from the surface and intracellular of labeled cells across multiple cycles, allowing for more accurate and efficient molecular profiling of their complex phenotypes. Herein, the versatility of this technology is demonstrated for the screening of glioblastoma markers in patient‐derived human glioblastoma organoids while conserving their viability. It is anticipated that the findings and applications of this work can be broadly translated into investigating physiological developments in other organoid systems.
A bioorthogonal click chemistry technology to display the first demonstration of multiplexed cyclic imaging of live and fixed patient‐derived glioblastoma tumor organoids. This approach enhances molecular profiling by quenching fluorescent signals from the surface and intracellular of labeled cells across multiple cycles, allowing for more efficient profiling of their complex phenotypes.
Serial tissue sampling has become essential in guiding modern targeted and personalized cancer treatments. An alternative to image guided core biopsies are fine needle aspirates (FNA) that yield ...cells rather than tissues but are much better tolerated and have lower complication rates. The efficient pathway analysis of such cells in the clinic has been difficult, time consuming and costly. Here we develop an antibody-DNA barcoding approach where harvested cells can be rapidly re-stained through the use of custom designed oligonucleotide-fluorophore conjugates. We show that this approach can be used to interrogate drug-relevant pathways in scant clinical samples. Using the PI3K/PTEN/CDK4/6 pathways in breast cancer as an example, we demonstrate how analysis can be performed in tandem with trial enrollment and can evaluate downstream signaling following therapeutic inhibition. This approach should allow more widespread use of scant single cell material in clinical samples.
This study analyzed Spanish teachers’ behavior and the transmission of gender stereotypes in physical education classes in secondary schools in Seville, southern Spain. We observed 48 physical ...education lessons given by four Spanish teachers (two men and two women). Descriptive codes, which were generated iteratively, were clustered, categorized, integrated, recoded, and re-categorized. They allowed us to identify five major themes related to the transmission of gender stereotypes of teachers: the channel of communication, use of demonstrations, task organization, disciplinary measures, and questions asked. We used a coding sheet as well as audio and video recordings to register the categories. The Mann–Whitney test produced significance levels lower than .05, resulting in the null hypothesis being rejected. Sexist behaviors were found in the channel of communication, the students chosen for demonstrations, task organization (the groupings), and in the monitoring of group discipline. However, we found no difference in task organization (material and space) or quantity of questions asked.
Conventional CD4
+
T (Tconv) lymphocytes play important roles in tumor immunity; however, their contribution to tumor elimination remains poorly understood. Here, we describe a subset of ...tumor-infiltrating Tconv cells characterized by the expression of CD39. In several mouse cancer models, we observed that CD39
+
Tconv cells accumulated in tumors but were absent in lymphoid organs. Compared to tumor CD39
−
counterparts, CD39
+
Tconv cells exhibited a cytotoxic and exhausted signature at the transcriptomic level, confirmed by high protein expression of inhibitory receptors and transcription factors related to the exhaustion. Additionally, CD39
+
Tconv cells showed increased production of IFN
$\gamma $
γ
, granzyme B, perforin and CD107a expression, but reduced production of TNF. Around 55% of OVA-specific Tconv from B16-OVA tumor-bearing mice, expressed CD39. In vivo CTLA-4 blockade induced the expansion of tumor CD39
+
Tconv cells, which maintained their cytotoxic and exhausted features. In breast cancer patients, CD39
+
Tconv cells were found in tumors and in metastatic lymph nodes but were less frequent in adjacent non-tumoral mammary tissue and not detected in non-metastatic lymph nodes and blood. Human tumor CD39
+
Tconv cells constituted a heterogeneous cell population with features of exhaustion, high expression of inhibitory receptors and CD107a. We found that high CD4 and ENTPD1 (CD39) gene expression in human tumor tissues correlated with a higher overall survival rate in breast cancer patients. Our results identify CD39 as a biomarker of Tconv cells, with characteristics of both exhaustion and cytotoxic potential, and indicate CD39
+
Tconv cells as players within the immune response against tumors.
Intradialytic hypotension is common complication in stage 5 chronic kidney disease patients on hemodialysis. Incidence ranges from 15 to 30%. These patients have levocarnitine deficiency. A ...randomized, placebo‐controlled quadruple‐blinded trial was designed to demonstrate the levocarnitine efficiency on intradialytic hypotension prevention. Patients were randomized into four groups, to receive levocarnitine or placebo. During the intervention period, levocarnitine and placebo was administered 0 and 30 min before each hemodialysis session, respectively. During the trial, 33 patients received 1188 hemodialysis sessions. We identified 239 (21.3%) intradialytic hypotension episodes. The intradialytic hypotension episodes were less frequent in the levocarnitine group (9.3%, 60 IH events) (P < 0.001). Hemodialysis is frequently perplexed by intradialytic hypotension episodes. Levocarnitine supplementation before each hemodialysis session efficiently diminishes the intradialytic hypotension episodes. This is a new application method that must be considered and explored.
Purpose
This study was designed to present the 5-year results of patients with multicentric breast cancer who underwent sentinel lymph node biopsy (SLNB) in a single institution.
Methods
Between June ...1999 and December 2007, 337 patients with multicentric breast cancer and a clinically negative axilla underwent lymphatic mapping by a single periareolar/peritumoral (
n
= 306) or a double peritumoral or subdermal injection (
n
= 31) of
99m
Tc-HSA nanocolloids. The sentinel lymph node (SLN) was evaluated by intraoperative frozen section and axillary dissection was performed only in cases of positive SLNB.
Results
The median age of the patients was 48 (range, 22–81) years. The mean number of hot spots identified was 1.4 in the whole series, 1.3 in patients who received a single injection, and 1.7 in those who received a double injection (
P
< 0.001). The mean number of removed SLNs was 1.7 (median, 1; range, 1–7) with an identification rate of 100%. A total of 138 patients with negative SLNB (
n
= 134) or isolated tumor cells in the SLN (
n
= 4) did not receive completion axillary lymph node dissection (CALND). In these latter patients, a total of 27 events (19.5%) occurred with 3 patients (2.2%) developing axillary recurrences after a median follow-up of 5 years (range, 17–134 months).
Conclusions
Axillary lymph node reappearance was infrequent among patients with multicentric breast cancer, having negative SLNB and no CALND. We recommend SLNB as the standard procedure for nodal staging in patients with multicentric breast cancer and a clinically negative axilla.
PDF
