Carlotta Rossi - Academia.edu (original) (raw)

Papers by Carlotta Rossi

Research paper thumbnail of Co-cultures of enterocytes and hepatocytes for retinoid transport and metabolism

Toxicology in Vitro, 2012

Dietary retinoid bioavailability involves the interplay of the intestine (transport and metabolis... more Dietary retinoid bioavailability involves the interplay of the intestine (transport and metabolism) and the liver (secondary metabolism). To reproduce these processes in vitro, differentiated human intestinal Caco-2/TC7 cells were co-cultured with two hepatocyte cell lines. Murine 3A cells and the more highly differentiated human HepaRG hepatocytes were both shown to respond to b-carotene (BC) and retinol (ROH) treatment by secreting Retinol Binding Protein 4 (RBP4). In co-culture experiments, Caco-2/TC7 were differentiated on filter inserts and transferred for the time of the experiment to culture wells containing confluent 3A or differentiated HepaRG cells. Functionality of the co-cultures was assayed using as endpoints the retinol-dependent secretion of RBP4 and the retinoic acid-dependent induction of CYP26A1 in hepatocytes. BC and ROH added to intestinal Caco-2/TC7 induced a reduction in intracellular RBP4 levels in the underlying hepatocytes and its secretion into the medium. HepaRG hepatocytes were also shown to up-regulate the expression of CYP26A1 mRNA in response to retinoid treatment. This in vitro model represents a useful tool to analyze the absorption and metabolism of retinoids and could be further developed to investigate other dietary compounds and molecules of pharmacological interest.

Research paper thumbnail of Variable costs of ICU patients: a multicenter prospective study

Intensive Care Medicine, 2006

Objective To analyze the costs of treating critically ill patients. Design and setting Multicente... more Objective To analyze the costs of treating critically ill patients. Design and setting Multicenter, observational, prospective, cohort, bottom-up study on variable costs in 51 ICUs. Patients and participants A total of 1,034 patients aged over 14years who either spent less than 48h in the ICU or had multiple trauma, major abdominal surgery, ischemic stroke, chronic obstructive pulmonary disease, cardiac failure, isolated head injury, acute lung injury/adult respiratory distress syndrome (ALI/ARDS), nontraumatic intracranial hemorrhage or coronary surgery. Interventions Data recorded for each patient: length of ICU stay, and cost in euros of all diagnostic and therapeutic procedures, drugs and equipment used, and consultations by physicians from other units. To express cost-efficiency we calculated for each diagnostic group the cost per surviving patient (expenditure for all patients/number of surviving patients) and money loss per patient (expenditure for patients who died/total number of patients).

Research paper thumbnail of The relationship between labour cost per patient and the size of intensive care units: a multicentre prospective study

Intensive Care Medicine, 2003

Research paper thumbnail of A protocol for differentiation of human intestinal Caco-2 cells in asymmetric serum-containing medium

The human intestinal Caco-2 cell line still represents the best available in vitro model of absor... more The human intestinal Caco-2 cell line still represents the best available in vitro model of absorptive enterocytes, despite its origin from a colon adenocarcinoma. Caco-2 cells seeded on filter inserts undergo in culture a process of spontaneous differentiation that leads to the formation, after two to three weeks, of a monolayer of polarized cell, coupled by tight junctions and expressing several morphological and functional features of small intestinal enterocytes. The medium normally used for differentiation of Caco-2 cells contains a supplement of foetal bovine serum (FBS) in both the apical (AP) and basolateral (BL) compartments. The use of FBS as cell culture media supplement has been frequently and increasingly questioned on scientific and also on ethical grounds. We have shown that addition of serum only to the BL medium (asymmetric protocol) appears to be sufficient to allow differentiation of Caco-2 cells, as monitored by morphology, monolayer permeability and alkaline phosphatase activity, compared to standard conditions using 10% FBS supplement in both AP and BL media (asymmetric protocol). Although not eliminating the use of FBS, its addition only in the BL medium results in more physiological conditions for differentiation and in a significant reduction of its use.

Research paper thumbnail of Co-cultures of enterocytes and hepatocytes for retinoid transport and metabolism

Toxicology in Vitro, 2012

Dietary retinoid bioavailability involves the interplay of the intestine (transport and metabolis... more Dietary retinoid bioavailability involves the interplay of the intestine (transport and metabolism) and the liver (secondary metabolism). To reproduce these processes in vitro, differentiated human intestinal Caco-2/TC7 cells were co-cultured with two hepatocyte cell lines. Murine 3A cells and the more highly differentiated human HepaRG hepatocytes were both shown to respond to b-carotene (BC) and retinol (ROH) treatment by secreting Retinol Binding Protein 4 (RBP4). In co-culture experiments, Caco-2/TC7 were differentiated on filter inserts and transferred for the time of the experiment to culture wells containing confluent 3A or differentiated HepaRG cells. Functionality of the co-cultures was assayed using as endpoints the retinol-dependent secretion of RBP4 and the retinoic acid-dependent induction of CYP26A1 in hepatocytes. BC and ROH added to intestinal Caco-2/TC7 induced a reduction in intracellular RBP4 levels in the underlying hepatocytes and its secretion into the medium. HepaRG hepatocytes were also shown to up-regulate the expression of CYP26A1 mRNA in response to retinoid treatment. This in vitro model represents a useful tool to analyze the absorption and metabolism of retinoids and could be further developed to investigate other dietary compounds and molecules of pharmacological interest.

Research paper thumbnail of Variable costs of ICU patients: a multicenter prospective study

Intensive Care Medicine, 2006

Objective To analyze the costs of treating critically ill patients. Design and setting Multicente... more Objective To analyze the costs of treating critically ill patients. Design and setting Multicenter, observational, prospective, cohort, bottom-up study on variable costs in 51 ICUs. Patients and participants A total of 1,034 patients aged over 14years who either spent less than 48h in the ICU or had multiple trauma, major abdominal surgery, ischemic stroke, chronic obstructive pulmonary disease, cardiac failure, isolated head injury, acute lung injury/adult respiratory distress syndrome (ALI/ARDS), nontraumatic intracranial hemorrhage or coronary surgery. Interventions Data recorded for each patient: length of ICU stay, and cost in euros of all diagnostic and therapeutic procedures, drugs and equipment used, and consultations by physicians from other units. To express cost-efficiency we calculated for each diagnostic group the cost per surviving patient (expenditure for all patients/number of surviving patients) and money loss per patient (expenditure for patients who died/total number of patients).

Research paper thumbnail of The relationship between labour cost per patient and the size of intensive care units: a multicentre prospective study

Intensive Care Medicine, 2003

Research paper thumbnail of A protocol for differentiation of human intestinal Caco-2 cells in asymmetric serum-containing medium

The human intestinal Caco-2 cell line still represents the best available in vitro model of absor... more The human intestinal Caco-2 cell line still represents the best available in vitro model of absorptive enterocytes, despite its origin from a colon adenocarcinoma. Caco-2 cells seeded on filter inserts undergo in culture a process of spontaneous differentiation that leads to the formation, after two to three weeks, of a monolayer of polarized cell, coupled by tight junctions and expressing several morphological and functional features of small intestinal enterocytes. The medium normally used for differentiation of Caco-2 cells contains a supplement of foetal bovine serum (FBS) in both the apical (AP) and basolateral (BL) compartments. The use of FBS as cell culture media supplement has been frequently and increasingly questioned on scientific and also on ethical grounds. We have shown that addition of serum only to the BL medium (asymmetric protocol) appears to be sufficient to allow differentiation of Caco-2 cells, as monitored by morphology, monolayer permeability and alkaline phosphatase activity, compared to standard conditions using 10% FBS supplement in both AP and BL media (asymmetric protocol). Although not eliminating the use of FBS, its addition only in the BL medium results in more physiological conditions for differentiation and in a significant reduction of its use.