86 research outputs found
Differential responsiveness of proliferation and cytokeratin release to stripped serum and oestrogen in the human breast cancer cell line, MCF-7.
No full textIn vitro research into hormone sensitivity and the relation to proliferation of cytokeratin release from cancer cells is scarce. Therefore, we examined the stimulation of proliferation and the release of cytokeratins in a breast cancer cell culture model. Cell growth was stimulated by 17 beta-oestradiol (10(-11) M), stripped serum (10%) and by the two together. Cytokeratin release was stimulated only by stripped serum, oestradiol having no effect. After long incubation periods (> 12 h), cytokeratin release also commenced in the control and oestradiol treatments. Release rate versus time analysis suggested that there are two different release processes. Cytokeratin release was first stimulated at a stripped serum concentration approximately 100 times lower than that which initiated proliferation. Pharmacological alteration of proliferation with cordyceptin resulted in growth changes without alterations in cytokeratin release. We conclude that cytokeratin release in these cells is unrelated to proliferation, independent of oestrogen action and probably in some way related to growth factor receptor function
Telomere length and telomerase activity in T cells are biomarkers of high-performing centenarians
No full textIt is generally recognized that the function of the immune system declines with increased age and one of the major immune changes is impaired T-cell responses upon antigen presentation/stimulation. Some "high-performing" centenarians (100+ years old) are remarkably successful in escaping, or largely postponing, major age-related diseases. However, the majority of centenarians ("low-performing") have experienced these pathologies and are forced to reside in long-term nursing facilities. Previous studies have pooled all centenarians examining heterogeneous populations of resting/unstimulated peripheral blood mononuclear cells (PBMCs). T cells represent around 60% of PBMC and are in a quiescence state when unstimulated. However, upon stimulation, T cells rapidly divide and exhibit dramatic changes in gene expression. We have compared stimulated T-cell responses and identified a set of transcripts expressed in vitro that are dramatically different in high- vs. low-performing centenarians. We have also identified several other measurements that are different between high- and low-performing centenarians: (a) The amount of proliferation following in vitro stimulation is dramatically greater in high-performing centenarians compared to 67- to 83-year-old controls and low-performing centenarians; (b) telomere length is greater in the high-performing centenarians; and (c) telomerase activity following stimulation is greater in the high-performing centenarians. In addition, we have validated a number of genes whose expression is directly related to telomere length and these are potential fundamental biomarkers of aging that may influence the risk and progression of multiple aging conditions
Release of the aspartyl protease cathepsin D is associated with and facilitates human breast cancer cell invasion
No full textData concerning the hormone sensitivity of the release and role of the aspartyl protease cathepsin D in tumor proliferative and invasive processes have been contradictory. To clarify the mechanisms of its release and role we first studied the contribution of estradiol and stripped serum to the time course and kinetics of cathepsin D release, proliferation, and invasion in parallel in the MCF-7 in vitro breast cancer cell culture model. Both estradiol and stripped serum independently stimulated both proliferation and cathepsin D release. However, the dose-response of estradiol and stripped serum-dependent stimulated release were similar to those for invasion and differed from those for proliferation: cathepsin D release and invasion were first stimulated at a stripped serum concentration more than 10-fold lower than that which initiated proliferation and had half stimulation constants almost 10-fold lower than those for proliferation. These results demonstrate that cathepsin D release is not related in any direct way to proliferation. The effect of the reduction of cathepsin D activity or release on in vitro invasion was also measured: both the inhibition of secreted cathepsin D activity by a specific inhibitor, diazoacetyl-DL-Nle-OMe, and the reduction of cathepsin D release by antisense oligonucleotides against its translation start site reduced cellular in vitro invasion without affecting proliferation. Cathepsin D release and activity are concluded to be directly involved in the process of invasion
Association of pS2 (TFF1) release with breast tumor proliferative rate: in vitro and clinical evidence
No full textAlthough cytosolic expression of the protein pS2 (TFF1) is considered to be a marker of oestrogen receptor (OR) function, there exists some clinical data to suggest an inverse relationship of cytosolic pS2 to tumour proliferation. Although secreted from breast cancer cells, the relationship of pS2 secretion to tumour natural history has been little studied. The mechanisms and kinetics of pS2 release and its relation to tumour cell proliferation were studied in a human breast cancer cell line MCF-7 and verified in a preliminary clinical study. Stimulation by stripped serum or oestradiol resulted in parallel increases of proliferation and pS2 release in both time course and dose-response experiments. Direct pharmacological alterations of proliferation were followed by identical changes in pS2 release. The relationship between serum pS2 levels and tumour proliferative activity when analysed as a function of steroid status showed a slope of 0.56 in OR+ vs. 0.19 in OR- tumours. It is concluded that pS2 release from breast cancer cells is associated with their proliferation and measurement of serum pS2 levels might be a good predictor of tumour proliferative state and could permit noninvasive monitoring of this tumour parameter
Cytokeratins and proliferation in breast cancer patients.
No full textTissue polypeptide antigen (TPA) was studied in 242 sera and 165 tumor cell cytosols (both evaluations in 67 cases) of breast cancer patients, for which proliferative activity, determined by the TLI technique, was also available. The TPA serum and tumor cell cytosol median values (utilized for measure analysis as cut-off) were 70 U/1 and 377 U/mg cytosol protein, respectively. High serum TPA levels were associated with unfavourable clinicopathological characteristics whereas a higher tumor cell cytosol TPA level was associated with better cytohistological tumor differentiation. Furthermore, no overall correlation was found between serum and tumor cell cytosol TPA levels or between their levels and TLI. When analyzing cases in which serum and tumor cell cytosol TPA values were higher than 100 U/l and 500 U/mg cytosol protein, respectively (n = 28), serum TPA was positively associated with TLI (slope = 12.3 r = 0.55, p < 0.01), while cytosolic TPA resulted negatively associated with TLI (slope = -87.4 r = 0.41, p < 0.01). Finally, a strong inverse relationship between cytosolic and serum TPA (p < 0.0005) became evident. We suggest that TPA could represent a serum marker for tumor cell proliferation in specific patient subgroups with original high serum and/or cytosol TPA expression
Cathepsin D, an acidic aspartyl protease, is associated with and facilitates human cancer cell invasion
No full textQuantitative mitochondrial DNA copy number determination using droplet digital PCR with single cell resolution
No full textMitochondria are involved in a number of diverse cellular functions, including energy production, metabolic regulation, apoptosis, calcium homeostasis, cell proliferation, and motility, as well as free radical generation. Mitochondrial DNA (mtDNA) is present at hundreds to thousands of copies per cell in a tissue-specific manner. mtDNA copy number also varies during aging and disease progression and therefore might be considered as a biomarker that mirrors alterations within the human body. Here, we present a new quantitative, highly sensitive droplet digital PCR (ddPCR) method, droplet digital mitochondrial DNA measurement (ddMDM), to measure mtDNA copy number not only from cell populations but also from single cells. Our developed assay can generate data in as little as 3 h, is optimized for 96-well plates, and also allows the direct use of cell lysates without the need for DNA purification or nuclear reference genes. We show that ddMDM is able to detect differences between samples whose mtDNA copy number was close enough as to be indistinguishable by other commonly used mtDNA quantitation methods. By utilizing ddMDM, we show quantitative changes in mtDNA content per cell across a wide variety of physiological contexts including cancer progression, cell cycle progression, human T cell activation, and human aging
Variability in Productive and Biochemical Traits of Vicia faba L. Landraces from Apulia Region (South Italy)
No full textThe faba bean (Vicia faba L. var. major) is a pulse that is garnering attention for its chemical composition, which makes it suitable for a healthy diet. The Apulian germplasm is rich in local accessions at risk of genetic erosion, which need evaluating and promoting. Thirteen Vicia faba local Landraces have been analyzed in relation to their productivity and their chemical and biochemical characteristics: their protein, total phenol, total flavonoid, condensate tannin and L-DOPA levels. The results showed great variability—above all in the thousand-seeds weight and in their content of proteins and L-DOPA. Among the accessions evaluated, the two collected from the most southern area of the region (FV12-FV10) were particularly promising—both for their good biochemical traits and, especially, for the higher L-DOPA content (0.46 and 0.49 g 100 g−1 d.m., respectively), even when expressed in terms of yield per plant (116.3 and 153.0 mg plant−1 d.m., respectively). © 2023 by the author
Early CAR− CD4+ T‐lymphocytes recovery following CAR‐T cell infusion: A worse outcome in diffuse large B cell lymphoma
No full textAbstract CAR− CD4+ T cell lymphopenia is an emerging issue following CAR‐T cell therapy. We analyzed the determinants of CD4+ T cell recovery and a possible association with survival in 31 consecutive patients treated with commercial CAR‐T for diffuse large B‐cell (DLBCL) or mantle cell lymphoma. Circulating immune subpopulations were characterized through multiparametric‐flow cytometry. Six‐month cumulative incidence of CAR− CD4+ T cell recovery (≥200 cells/μL) was 0.43 (95% confidence interval [CI]: 0.28–0.65). Among possible determinants of CD4+ T cell recovery, we recognized infusion of a 4‐1BB product (tisagenlecleucel, TSA) in comparison with a CD28 (axicabtagene/brexucabtagene, AXI/BRX) (hazard ratio [HR] [95% CI]: 5.79 [1.16–24.12] p = 0.016). Higher CD4+ T cell counts resulted with TSA at month‐1, ‐2 and ‐3. Moderate‐to‐severe infections were registered with prolonged CD4+ T cell lymphopenia. Early, month‐1 CD4+ T cell recovery was associated with a worse outcome in the DLBCL cohort, upheld in a multivariate regression model for overall survival (HR: 4.46 [95% CI: 1.12–17.71], p = 0.03). We conclude that a faster CAR− CD4+ T cell recovery is associated with TSA as compared to AXI/BRX. Month‐1 CAR− CD4+ T cell subset recovery could represent a “red flag” for CAR‐T cell therapy failure in DLBCL patients
Camelina sativa (L. Crantz) Fresh Forage Productive Performance and Quality at Different Vegetative Stages: Effects of Dietary Supplementation in Ionica Goats on Milk Quality
No full textThe research meant to study the productive performances of Camelina sativa and the effects of feeding Camelina fresh forage harvested during five phenological stages (I: main stem elongation; II: maximum stem elongation: III: inflorescence appearance; IV: flowering; V: fruit set visible) on the yield, chemical composition and fatty acid profile of milk from autochthonous Ionica goats. Goats were randomly assigned to two groups (n = 15) that received a traditional forage mixture (Control) or Camelina forage harvested at different stages (CAM). The field experiment was conducted in two years; no significant differences between years were recorded for any of the Camelina production traits. The total biomass increased (p < 0.05) from phase I (1.4 t/ha) to phase V (5.2 t/ha). The distribution of stem, leaves and pod also changed during growth, showing a significant increase of stem from 40.8 to 45.6% and of pod from 0 to 19.4%, whereas leaves decreased from 59.2 to 35.1%. The milk yield and chemical composition were unaffected by the diet, while supplementation with Camelina forage increased milk CLA content (on average 1.14 vs. 0.78%). A markedly higher concentration of PUFAs was found in milk from goats fed Camelina harvested during the last three phenological stages. The index of thrombogenicity of milk from the CAM fed goats was significantly lower compared to the control group. In conclusion, Camelina sativa is a multi-purpose crop that may be successfully cultivated in Southern Italy regions and used as fresh forage for goat feeding. Milk obtained from Camelina fed goats showed satisfactory chemical and fatty acid composition, with potential benefits for human health
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