Abstract Dendritic cells (DCs), specialized antigen-presenting cells bridging innate and adaptive immunity, play a crucial role in determining specific immune response to tumors. Because of their ...potent immunoregulatory capacities, DCs have been exploited in anticancer vaccination, with limited success thus far. This pilot study compared low-dose interleukin (IL)-4 and IL-12 prepared by sequential kinetic activation (SKA) with standard doses of the same recombinant human cytokines on functional activity of ex vivo –generated monocyte-derived (Mo) DCs from colon carcinoma patients and normal subjects. MoDCs were exposed to medium alone, SKA-IL-4 (0.5 fg/ml), or SKA-IL-12 (2 fg/ml), alone or consecutively combined, in parallel with rhIL-4 (50 ng/ml) and rhIL-12 (1 ng/ml). Primary allogeneic one-way mixed lymphocyte reaction (MLR) was the end point to assess in vitro T-lymphocyte proliferation in response to MoDCs, and secreted IL-12p70 and interferon-γ in MLR supernatants measured by ELISA to assay for T-helper 1–promoting MoDC phenotype. No single agent enhanced the compromised allostimulatory activity of MoDCs from colon cancer patients, unlike healthy donors. However, MoDCs from nonmetastatic colon cancer patients, after sequential exposure to SKA-IL-4 (48 hours) and SKA-IL-12 (24 hours), displayed increased T-cell stimulatory capacity by MLR and acquired driving T-helper 1 polarization activity, although less markedly than the effects induced by recombinant human cytokines or found in normal subjects. These results point to an immunomodulatory capacity of low-dose SKA-IL-4 and SKA-IL-12 and encourage further investigation to provide clues for the rational development of new and more effective immunotherapeutic strategies against cancer.
Since interferon-γ (IFN-γ) tunes both innate and adaptive immune systems, it was expected to enter clinical practice as an immunomodulatory drug. However, the use of IFN-γ has been limited by its ...dose-dependent side effects. Low-dose medicine, which is emerging as a novel strategy to treat diseases, might circumvent this restriction. Several clinical studies have proved the efficacy of therapies with a low dose of cytokines subjected to kinetic activation, while no in vitro data are available. To fill this gap, we investigated whether low concentrations, in the femtogram range, of kinetically activated IFN-γ modulate the behavior of Jurkat cells, a widely used experimental model that has importantly contributed to the present knowledge about T cell signaling. In parallel, IFN-γ in the nanogram range was used and shown to activate Signal transducer and activator of transcription (STAT)-1 and then to induce suppressor of cytokine signaling-1 (SOCS-1), which inhibits downstream signaling. When added together, femtograms of IFN-γ interfere with the transduction cascade activated by nanograms of IFN-γ by prolonging the activation of STAT-1 through the downregulation of SOCS-1. We conclude that femtograms of IFN-γ exert an immunomodulatory action in Jurkat cells.
The Children's Sleep Habits Questionnaire (CSHQ) is a parent-report questionnaire used to examine sleep behavior in children. Linguistic adaptation of CSHQ into several languages and/or psychometric ...analysis of reliability have been published.
Our aim was to translate the original 33-items CSHQ from English to Italian and to examine its reliability for use in 4-10 years-old children of Italy. After translation and back-translation procedure of the original CSHQ, the Italian CSHQ (CSHQ-IT) was administered to 69 mothers of healthy children. Reliability of CSHQ-IT was examined by the internal consistency of the scale (using the Cronbach's alpha coefficient), and by the test-retest analysis obtained by asking mothers who had completed the CSHQ-IT at baseline to re-complete it after a two-week interval (measured with the Lin's Concordance Correlation Coefficient, CCC). The CSHQ-IT showed adequate internal consistency (Cronbach's alpha = 0.81 for the total scale). The total CSHQ-IT score showed a strong correlation in retests (CCC 0.87; 95% Confidence Interval, 0.80; 0.92).
CSHQ-IT is a valuable tool for evaluating sleep behavior in children 4-10 years-old in Italy. Future research should be implemented to provide definitive validity of CSHQ-IT in children with sleep-disordered breathing.
Natural killer (NK) cells are innate immune-system lymphocytes capable of killing tumor cells. They secrete cytokines, including interferon (IFN)-γ, which participate in shaping the initial ...inflammatory and downstream adaptive immune responses. Its potent immunoregulatory action means that IFN-γ might be beneficial in cases of tumor rejection, but its severe side-effects limit clinical applications. This pilot study compared low-dose IFN-γ prepared by sequential-kinetic-activation (SKA), with standard-dose recombinant (r) IFN-γ, in terms of ex-vivo cytotoxic activity of peripheral blood (PB)-NK cells from colorectal carcinoma (CRC) patients. This was tested against the NK-sensitive K562 cell line and the less-sensitive human CRC Caco-2 and HT-29 cell lines. Twenty primitive non-metastatic CRC patients, five metastatic CRC patients, and thirteen healthy donors were enrolled. PB lymphocytes (PBL) were exposed to medium alone, SKA-IFN-γ (0.25fg/ml) or rIFN-γ (1ng/ml). NK-cell cytolytic activity was examined via short-term 51Cr-release. Pretreatment of PBL from non-metastatic patients with SKA-IFN-γ caused a significant increase in NK-cell cytotoxicity, compared to those from normal donors, although less markedly than pretreatment with rIFN-γ against all three cell lines. In contrast, PBL from metastatic CRC patients displayed significantly decreased NK-cell activity and responsiveness to both rIFN-γ and SKA-IFN-γ treatments. These results demonstrate in principle the immunomodulatory capacity of low-dose SKA-IFN-γ, and might open the door to the possibility of generating a novel, safe, and feasible approach to enhancing NK-cell antitumor activity in early-stage CRC patients.
To study the efficacy of low-dose SKA Progesterone and IL-10 in modulating the inflammatory pathway in endometriosis.
Experimental basic science study.
Reproductive biology laboratory.
Immortalized ...human endometriotic epithelial cells (12Z) derived from active red peritoneal lesions, immortalized human endometriotic stromal cells (22B) derived from active red peritoneal lesions and immortalized human endometrial cell line T-Hesc (ATCC collection).
Cells were treated with SKA-Progesterone and SKA-IL10 at low doses (10pg/ml and 10fg/ml respectively). Medroxyprogesterone 17-acetate (MPA) was used at a dose of 10μM as reference treatment.
Modulation of HSD17B1 levels by WB analysis after low-dose SKA Progesterone and MPA; Modulation of IKBα protein levels and NF-kB p65 nuclear levels by WB analysis after low-dose SKA-Progesterone, low-dose SKA-IL10, low-dose SKA-Progesterone and low-dose SKA-IL10 (combined treatment), MPA.
Low-dose SKA Progesterone was effective in the inhibition of HSD17B1 expression in endometriotic epithelial (12Z) and stromal (22B) cell lines. Low-dose of SKA Progesterone and low-dose of SKA-IL10 inhibit NF-kB p65 nuclear localization and DNA binding in endometriotic epithelial (12Z) cells, stromal (22B) cells line and in endometrial cell line T-Hesc. The combined treatment showed an additive effect, namely increasing the inhibition of nuclear localization of NF-kB p65 and DNA binding as result of single treatments.
Our data suggest that the use of a combination of low-dose SKA Progesterone and IL-10 may represent an opportunity for the development of new therapies in the clinical management of endometriosis.
•A study on endometriotic cells treated with SKA-Progesterone and SKA-IL10 at low doses is proposed.•Low-dose SKA Progesterone inhibits HSD17B1 expression in endometriotic cell lines.•Low-dose Progesterone and IL10 inhibit NF-kB p65 nuclear localization.•The use of a combination of low-dose SKA Progesterone and IL-10 may represent a new therapy of endometriosis.
In this paper we present an innovative approach to jointly use analytical and numerical methods to investigate the effects of high permittivity helmets in magnetic resonance scanners. The proposed ...methodology and the obtained results provide an increased comprehension of the field-sample interaction that can be fruitfully used to support the design of dielectric helmets in magnetic resonance imaging.
In this work we present a new analytical framework to investigate how the presence of an insulating layer, such as the distance between a high-permittivity helmet and the body tissues can impact the ...field distribution in the region of interest, and the choice of the optimum permittivity to this purpose. The framework is based on a recent reformulation of the Mie scattering for a layered sphere and is validated with a numerical method based on the finite difference time domain.
Objective: To investigate the activity-based prospective memory performance in patients with insomnia, divided, on the basis of actigraphic evaluation, into sleep onset, maintenance, mixed and ...negative misperception insomnia. Methods: A total of 153 patients with insomnia (I, 83 females, mean age + SD = 41.37 + 16.19 years) and 121 healthy controls (HC, 78 females, mean age + SD = 36.99 + 14.91 years) wore an actigraph for one week. Insomnia was classified into sleep onset insomnia (SOI), maintenance insomnia (MaI), mixed insomnia (MixI) and negative misperception insomnia (NMI). To study their activity-based prospective memory performance, all the participants were required to push the actigraph event marker button twice, at bedtime (task 1) and at get-up time (task 2). Results: Only patients with maintenance and mixed insomnia had a significantly lower accuracy in the activity-based prospective memory task at get-up time compared with the healthy controls. Conclusion: The results show that maintenance and mixed insomnia involve an impaired activity-based prospective memory performance, while sleep onset and negative misperception insomnia do not seem to be affected. This pattern of results suggests that the fragmentation of sleep may play a role in activity-based prospective memory efficiency at wake-up in the morning.
Prospective memory (PM) is essential in everyday life because it concerns the ability to remember to perform an intended action in the future. Individuals diagnosed with attention deficit ...hyperactivity disorder (ADHD) often show poor performance in PM. Because age can be confounding, we decided to test PM in ADHD patients (children and adults) and healthy controls (children and adults). We examined 22 children (four females; mean age = 8.77 ± 1.77) and 35 adults (14 females; mean age = 37.29 ± 12.23) with ADHD, in addition to 92 children (57 females; mean age = 10.13 ± 0.42) and 95 adults (57 females; mean age = 27.93 ± 14.35) as healthy controls. Each participant originally wore an actigraph around the non-dominant wrist and was requested to push the event-marker at get-up time. To assess the efficiency of PM performance, we calculated the time elapsing between the end of sleep in the morning and the pushing of the event-marker button. The results showed lower PM performance in ADHD participants, regardless of age. However, the differences between ADHD and control groups were more evident in the children group. Our data seem to confirm that PM efficiency is compromised in individuals diagnosed with ADHD regardless of age, and agree with the idea of considering the PM deficit as a neuropsychological marker of ADHD.
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