Automated Large-Scale Production of Paclitaxel Loaded Mesenchymal Stromal Cells for Cell Therapy Applications

Mesenchymal stromal cells (MSCs) prepared as advanced therapies medicinal products (ATMPs) have been widely used for the treatment of different diseases. The latest developments concern the possibility to use MSCs as carrier of molecules, including chemotherapeutic drugs. Taking advantage of their intrinsic homing feature, MSCs may improve drugs localization in the disease area. However, for cell therapy applications, a significant number of MSCs loaded with the drug is required. We here investigate the possibility to produce a large amount of Good Manufacturing Practice (GMP)-compliant MSCs loaded with the chemotherapeutic drug Paclitaxel (MSCs-PTX), using a closed bioreactor system. Cells were obtained starting from 13 adipose tissue lipoaspirates. All samples were characterized in terms of number/viability, morphology, growth kinetics, and immunophenotype. The ability of MSCs to internalize PTX as well as the antiproliferative activity of the MSCs-PTX in vitro was also assessed. The results demonstrate that our approach allows a large scale expansion of cells within a week; the MSCs-PTX, despite a different morphology from MSCs, displayed the typical features of MSCs in terms of viability, adhesion capacity, and phenotype. In addition, MSCs showed the ability to internalize PTX and finally to kill cancer cells, inhibiting the proliferation of tumor lines in vitro. In summary our results demonstrate for the first time that it is possible to obtain, in a short time, large amounts of MSCs loaded with PTX to be used in clinical trials for the treatment of patients with oncological diseases

Circadian blood pressure profile in patients with active Cushing's disease and after long-term cure

Hypertension is a major feature of Cushing's disease, with the attendant increase in the rate of cardiovascular events. The circadian blood pressure profile also impacts cardiovascular risk and a few studies have shown that patients with Cushing's syndrome do not present the expected nocturnal blood pressure decrease and, further, that this alteration persists in short-range disease remission. These studies were performed by conventional discontinuous ambulatory pressure monitoring, a technique not devoid of limitations. Aim of our study was the assessment of blood pressure and heart rate profile by beat-to-beat noninvasive monitoring in twelve patients with active Cushing's disease (9 women and 3 men, age 33.3 \ub1 2.36 years) and the assessment of its possible changes at short- (< 1 year) and long-term (2-3 years) follow-up after curative surgery. No nocturnal blood pressure dipping (i.e., decrease by 10% of daytime values) was observed in 50% of patients both during active hypercortisolism and within 1 year from surgery. Recovery of blood pressure dipping profile was detected at long-term follow-up in a minority of patients. Daytime heart rate was higher in patients with active Cushing's disease and decreased over time after cure. In conclusion, patients with Cushing's disease present absent nocturnal blood pressure dipping and abnormal heart rate values which do not resolve after short-term remission of hypercortisolism and show only partial improvement in the long run. These findings identify additional cardiovascular risk factors for patients cured of Cushing's disease

Mn(III) acetate induced addition of α-amidomalonic ester derivatives to conjugated olefins

Oxidation of diethyl α-amidomalonate derivatives 1 by Mn(III) acetate in the presence of conjugated olefins 2 leads to addition-oxidation products 3–5 in good to excellent yield. In neat furan and thiophene the 2-substitution products 5 are obtained. Side oxidation to α-acetoxymalonic derivatives is the main competitive reactio

1,5 vs. 1,6 Intramolecular homolytic aromatic substitution by vinyl radicals

Manganese(III) acetate oxidation of substituted diethyl benzylmalonates in the presence of alkynes affords tetrahydronaphthalene derivatives 3, 6, 7 and spiro[4,5]decatriene derivatives 4 or 5, through competitive 1,5 and 1,6 intramolecular aromatic substitution by vinyl radical

Synthesis of 2-oxy-3-(pyridinium-1'-yl)-1,4-naphthoquinone derivatives by iodine or hydrogen peroxide oxidation of 1,4-naphthoquinones in the presence of substituted pyridines.

2-Oxy-3-(pyridinium-1'-yl)-1,4-naphthoquinone betaines are obtained in moderate to good yield by oxidation of 1,4-naphthoquinones with iodine/MnO2 or hydrogen peroxide in the presence of substituted pyridines. Pyridinium-1,4-naphthoquinone iodides and 2,3-epoxy-2,3-dihydronaphthoquinone are suggested intermediate

Potentiating effect of galanin on GHRH-induced GH release : comparison between old and young subjects

The aim of our study was to investigate the effect of galanin on basal and GHRH-stimulated GH secretion in a healthy group of elderly subjects and in a healthy group of young subjects for comparison. Ten old subjects (mean age 75 +/- 1.15 years) and an equal number of healthy young volunteers (mean age 26 +/- 0.71 years) underwent three stimulation tests in random order. Galanin infusion for 60 minutes (10 micrograms/kg in 100 ml saline) failed to provoke an appreciable release of circulating GH in old subjects, while it induced a significant increase of plasma GH in the young adults; GHRH administration i.v. in bolus (100 micrograms in 1 ml saline) elicited a significant GH response in both groups; however, in the older group GH response was significantly (p < 0.05) lower than in the young adults. The administration of galanin (10 micrograms/kg in 100 ml saline as an i.v. infusion for 60 min) plus GHRH (100 micrograms in 1 ml saline i.v. in bolus), potentiated GH response in old and young subjects. The combined administration of two peptides was able to elicit a clear GH release even in the older subjects who were hyporesponsive/unresponsive to galanin and/or GHRH alone. In the elderly, plasma GH values observed after the combined stimuli overlapped with GH values observed after GHRH alone in the young adults. In conclusion, our study confirms that galanin has a synergic effect with GHRH on GH release both in younger and elderly subjects. Moreover, our data confirm an impaired responsivity to GHRH in the elderly and demonstrate that galanin is able to normalize response of somatotrophs to GHRH in old people