A multi-organ-on-chip to recapitulate the infiltration and the cytotoxic activity of circulating NK cells in 3D matrix-based tumor model

The success of immunotherapeutic approaches strictly depends on the immune cells interaction with cancer cells. While conventional in vitro cell cultures under-represent the complexity and dynamic crosstalk of the tumor microenvironment, animal models do not allow deciphering the anti-tumor activity of the human immune system. Therefore, the development of reliable and predictive preclinical models has become crucial for the screening of immune-therapeutic approaches. We here present an organ-on-chip organ on chips (OOC)-based approach for recapitulating the immune cell Natural Killer (NK) migration under physiological fluid flow, infiltration within a 3D tumor matrix, and activation against neuroblastoma cancer cells in a humanized, fluid-dynamic environment. Circulating NK cells actively initiate a spontaneous "extravasation " process toward the physically separated tumor niche, retaining their ability to interact with matrix-embedded tumor cells, and to display a cytotoxic effect (tumor cell apoptosis). Since NK cells infiltration and phenotype is correlated with prognosis and response to immunotherapy, their phenotype is also investigated: most importantly, a clear decrease in CD16-positive NK cells within the migrated and infiltrated population is observed. The proposed immune-tumor OOC-based model represents a promising approach for faithfully recapitulating the human pathology and efficiently employing the immunotherapies testing, eventually in a personalized perspective. An immune-organ on chip to recapitulate the tumor-mediated infiltration of circulating immune cells within 3D tumor model

Damage Control Surgery for Perforated Diverticulitis with Generalized Peritonitis: Better a Delayed Anastomosis than a Stoma Right Away.

Introduction. In the last decade, Damage Control Surgery (DCS) has been emerging as a feasible alternative management of patients with abdominal infection and sepsis. So far, there is no consensus about the role of DCS in perforated acute diverticulitis. In this study, we present the outcome from a multi-institutional series of patients presenting with grade III and IV Hinchey’s diverticulitis and managed by DCS. Method. All the partecipating centers were tertiary referral hospitals. A total of 34 patients with perforated diverticulitis treated with DCS and admitted between June 2015 and September 2017 were included in the study. During the first laparotomy, a limited resection of the diseased segment followed by a lavage and the application of an open abdomen technique was performed. After patient resuscitation, a second look was performed after 24/48 hours. Demographics, clinical, intra-operative and post-operative variables were carefully analyzed. Mortality, morbidity, and restoration of bowel continuity were the major outcomes of the study. Results. There were 15 male (44%) and 19 female (56%) with a mean age of 66,9 years (SD ± 12,7). Mean BMI was 28,42 Kg/m² (SD ± 3,33). Based on the severity of the disease, 13 cases (38%) were classified as Wasvary’s modified Hinchey’s stage III, and 21 cases (62%) as Hinchey IV. Mean Mannheim Peritonitis Index (MPI) was 25,12 (SD ± 6,28). In 22 (65%), ASA score was ≥ grade III. In all cases, the open abdomen was created by using a Negative Pressure Wound Therapy (NPWT) technique. At the second operation, twenty-four patients (71%) had a primary anastomosis, while 10 (29%) were treated with an end colostomy (Hartmann’s procedure). In 7/34 (21%) cases, a third look was needed. In 2/24 patients, a temporary loop ileostomy was required: both of them were closed in a second moment. Mortality rate was 12%. Overall morbidity rate was 53% (18/34). According to Claviend and Dindo classification, there were no grade I, 6/18 grade II, 1/18 grade IIIa, 5/18 grade IIIb and 2/18 grade IV. Reinterventions were required in 4/34 (12%): two for intestinal anastomosis dehiscence and two for abdominal wound dehiscence. Mean lenght of hospital stay was 21,9 days. Conclusion. DCS is feasable for patients with generalized peritonitis from perforated diverticulitis and it seems related to a higher rate of bowel reconstruction. Due to the open abdomen, it requires a stay in ICU with a prolonged mechanical ventilation but these same needs are often the burden of the majority of patients undergone surgery for a perforated diverticulitis, whatever the procedure is done

Damage control surgery for perforated diverticulitis with diffuse peritonitis: saves lives and reduces ostomy

Introduction Over the last decade, damage control surgery (DCS) has been emerging as a feasible alternative for the management of patients with abdominal infection and sepsis. So far, there is no consensus about the role of DCS for acute perforated diverticulitis. In this study, we present the outcome of a multi-institutional series of patients presenting with Hinchey's grade III and IV diverticulitis managed by DCS. Methods All the participating centers were tertiary referral hospitals. A total of 34 patients with perforated diverticulitis treated with DCS during the period 2011–2017 were included in the study. During the first laparotomy, a limited resection of the diseased segment was performed followed by lavage and use of negative pressure wound therapy (NPWT). After 24/48 h of resuscitation, patients returned to the operating room for a second look. Mortality, morbidity, and restoration of bowel continuity were the primary outcomes of the study. Results There were 15 males (44%) and 19 females (56%) with a mean age of 66.9 years (SD ± 12.7). Mean BMI was 28.42 kg/m2 (SD ± 3.33). Thirteen cases (38%) were Wasvary’s modified Hinchey's stage III, and 21 cases (62%) Hinchey's stage IV. Mean Mannheim Peritonitis Index (MPI) was 25.12 (SD ± 6.28). In 22 patients (65%), ASA score was ≥ grade III. Twenty-four patients (71%) had restoration of bowel continuity, while 10 (29%) patients had an end colostomy (Hartmann’s procedure). Three of these patients received a temporary loop ileostomy. One patient had an anastomotic leak. Mortality rate was 12%. Mean length of hospital stay was 21.9 days. At multivariate analysis, male gender (p = 0.010) and MPI (p = 0.034) correlated with a high percentage of Hartmann’s procedures. Conclusion DCS is a feasible procedure for patients with generalized peritonitis secondary to perforated diverticulitis, and it appears to be related to a higher rate of bowel reconstruction. Due to the open abdomen, stay in ICU with prolonged mechanical ventilation is required, but these aggressive measures may be needed by most patients undergoing surgery for perforated diverticulitis, whatever the procedure is done

Development of subnanomolar-affinity serotonin 5-HT4 receptor ligands based on quinoline structures

Two small series of quinoline derivatives were designed starting from previously published quinoline derivatives 7a and b in order to obtain information about their interaction with the 5-HT4R binding site. Initially, the structure of 7a and b was modified by replacing their basic moiety with that of partial agonist 4 (ML10302) or with that of reference ligand 6 (RS-67-333). Then, the aromatic moieties of 4-quinolinecarboxylates 7a, d-f, and h-k or 4-quinolinecarboxamides 7b, c, and g were modified into those of 2-quinolinecarboxamides 9a-e. Very interestingly, this structure-affinity relationship study led to the discovery of 7h-j as novel 5-HT4R ligands showing Ki values in the subnanomolar range. The structures of all these compounds contain the N-butyl-4-piperidinylmethyl substituent, which appear to behave as an optimized basic moiety in the interaction of these 4-quinolinecarboxylates with the 5-HT4R binding site. However, this basic moiety was ineffective in providing 5-HT4R affinity in the corresponding 4-quinolinecarboxamide 7g, but it did in 2-quinolinecarboxamide ligands 9c-e. Thus, a subtle interrelationship of several structural parameters appeared to play a major role in the interaction of the ligands with the 5-HT4R binding site. They include the kind of basic moiety, the position of the carbonyl linking group with respect to the aromatic moiety and its orientation, which could be affected by the presence of the intramolecular H-bond as in compounds 9c-e

Multivalent ligands for the serotonin 5-HT4 receptor

5-HT4 receptors are known to form constitutive dimers in membranes. To explore whether multivalency can enhance ligand interactions and/or efficacy in 5-HT4 receptors, the structure of the partial agonist ML10302 was modified with oligoIJethylene glycol) chains, thus generating, by a gradual approach, short and long tethered bivalent or tetravalent ligands and the corresponding spanner-linked monovalent controls. Both bivalent and tetravalent ligands displayed a 10–20-fold increase in binding affinity compared to appropriate controls, but no multivalent ligand showed greater binding energy than ML10302 itself. Furthermore, the direct assessment of receptor–Gs interaction and studies of cAMP signalling indicated that multivalency does not enhance the efficacy of ML10302

Synthesis and structure-activity relationship studies in serotonin 5-HT4 receptor ligands based on a benzo[de][2,6]naphthridine scaffold

A small series of serotonin 5-HT4 receptor ligands has been designed from flexible 2-methoxyquinoline compounds 7a,b by applying the conformational constraint approach. Ligands 7a,b and the corresponding conformationally constrained analogues 8aeg were synthesized and their interactions with the 5-HT4 receptor were examined by measuring both binding affinity and the ability to promote or inhibit receptoreG protein coupling. Ester derivative 7a and conformationally constrained compound 8b were demonstrated to be the most interesting compounds showing a nanomolar 5-HT4R affinity similar to that shown by reference ligands cisapride (1) and RS-23,597-190 (4). The result was rationalized by docking studies in term of high similarity in the binding modalities of flexible 7a and conformationally constrained 8b. The intrinsic efficacy of some selected ligands was determined by evaluating the receptoreG protein coupling and the results obtained demonstrated that the nature and the position of substituents play a critical role in the interaction of these ligands with their receptor