Proliferation and functional activation of endothelial cells within a tissue site of inflammation are regulated by humoral factors released by cells, such as T lymphocytes and monocytes, infiltrating the perivascular space. In the present study we investigated the effects of interleukin 3 (IL-3), an activated T lymphocyte-derived cytokine, on cultured human umbilical vein endothelial cells (HUVEC). Proliferative activity, evaluated both by estimation of the fraction of cells in the S phase and by direct cell count demonstrated that IL-3, at the dose of 25 ng/ml, enhances more than threefold both DNA synthesis and cell proliferation above baseline control conditions. Binding studies with radioiodinated ligand demonstrated that HU-VEC constitutively express a small number of IL-3 binding sites (-99 binding sites per cell, with an apparent Kd of 149 pM). Accordingly, molecular analysis showed the presence of transcripts for both a and ft subunits of the IL-3 receptor. Functional activation of endothelial cells was evaluated by the expression of the endothelial-leukocyte adhesion molecule 1 (ELAM-1) transcript and by leukocyte adhesion. The ELAM-1 gene transcript was clearly detectable 4 h after IL-3 addition and started to decrease after 12 h. Moreover, IL-3-induced ELAM-1 transcription was followed by enhanced adhesion of neutrophils and CD4' T cells to HUVEC. The findings that IL-3 can stimulate both proliferation and functional activation of endothelial cells suggest that this cytokine can be involved in sustaining the process of chronic inflammation. (J.

To elucidate the regulatory mechanisms of granulocyte-macrophage colony-stimulating factor (GM-CSF) production in human myeloid leukaemic cells we studied GM-CSF gene transcription, mRNA expression and GM-CSF secretion in human growth factor dependent M-07e cells. GM-CSF transcript was detected in cells cultured in the presence of interleukin-3 (IL-3). GM-CSF or mast cell growth factor (MGF), whereas it was undetectable in growth factor deprived cells. Growth factor re-addition induced, within 2 h, the appearance of GM-CSF mRNA. Nuclear run-on experiments demonstrated that the increase of GM-CSF mRNA levels depends on GM-CSF gene transcription. The simultaneous addition, to deprived cells, of the growth factor, and of cycloheximide (CHX) for 2 h inhibited GM-CSF mRNA expression, suggesting the requirement for newly made proteins for GM-CSF gene transcription. By means of the M-07e bioassay, which allows the detection of GM-CSF, IL-3 and MGF activities, and neutralizing antibodies to each of these factors, GM-CSF activity was detected in the cell-free extract of both IL-3- and MGF-sustained cells and of cells deprived for 24 h. This finding demonstrates that M-07e cells produce and store biologically active GM-CSF in response to both IL-3 and MGF. In contrast, analysis of the growth stimulatory activity present in the culture supernatants revealed that MGF, unlike IL-3, is able to induce the secretion of consistent amounts of GM-CSF. Taken together, our results suggest that, in M-07e cells, GM-CSF gene transcription and GM-CSF production are mediated, unlike its secretion, by mechanisms shared by IL-3 and MGF.

Deprescribing is a patient-centered process of medication withdrawal intended to achieve improved health outcomes through discontinuation of one or more medications that are either potentially harmful or no longer required. The objective of this study was to assess the perceptions of primary care physicians on deprescribing and potential barriers to deprescribing in the Local Health Authority (LHA) of Turin, Piedmont, Italy. Secondary objective was to evaluate educational needs of primary care physician. Cross sectional survey of primary care physicians working in the LHA of Turin, Piedmont, Italy. 439 GPs (71.3% of the total number of primary care physicians) attended an educational session related to deprescribing and were asked to anonymously answer a paper survey. Participants were asked to complete a previously published questionnaire about deprescribing and potential factors affecting the deprescribing process. A correlation coefficient was calculated to assess the association between physicians’ confidence in deprescribing and attitudes or barriers associated with deprescribing. Many GPs (71%) reported general confidence in their ability to deprescribe. Most respondents (83%) reported they were comfortable deprescribing preventive medications, however almost half expressed doubts regarding deprescribing when medication was initially prescribed by a colleague (45%) or when patient and/or caregiver supported the opportunity to continue the assumption (49%). Around a third of doctors maintain that the absence of strong evidence supporting deprescribing prevents them from considering it (38%), that they do not have the necessary time to effectively go through the process of deprescribing (29%), and that fear of possible effects due on withdrawal prevents them from deprescribing (31%). There was no strong correlation between physicians’ confidence and attitudes or barriers associated with deprescribing. The present study confirms that general practitioners sense the importance of deprescribing and feel prepared to face it managing communication with patients and caregivers, but find barriers when enacting the practice in a real-life context.

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