Efficacy of third-party chimeric antigen receptor modified peripheral blood natural killer cells for adoptive cell therapy of B-cell precursor acute lymphoblastic leukemia.

We developed an innovative and efficient, feeder-free culture method to genetically modify and expand peripheral blood-derived NK cells with high proliferative capacity, while preserving the responsiveness of their native activating receptors. Activated peripheral blood NK cells were efficiently transduced by a retroviral vector, carrying a second-generation CAR targeting CD19. CAR expression was demonstrated across the different NK-cell subsets. CAR.CD19-NK cells display higher antileukemic activity toward CD19+ cell lines and primary blasts obtained from patients with B-cell precursor ALL compared with unmodified NK cells. In vivo animal model data showed that the antileukemia activity of CAR.CD19-NK cell is superimposable to that of CAR-T cells, with a lower xenograft toxicity profile. These data support the feasibility of generating feeder-free expanded, genetically modified peripheral blood NK cells for effective "off-the-shelf" immuno-gene-therapy, while their innate alloreactivity can be safely harnessed to potentiate allogeneic cell therapy.

Zoledronic acid boosts γδ T-cell activity in children receiving αß+ T and CD19+ cell-depleted grafts from an HLA-haplo-identical donor.

We demonstrated that γδ T cells of patients given HLA-haploidentical HSCT after removal of αß+ T cells and CD19+ B cells are endowed with the capacity of killing leukemia cells after ex vivo treatment with zoledronic acid (ZOL). Thus, we tested the hypothesis that infusion of ZOL in patients receiving this type of graft may enhance γδ T-cell cytotoxic activity against leukemia cells. ZOL was infused every 28 d in 43 patients; most were treated at least twice. γδ T cells before and after ZOL treatments were studied in 33 of these 43 patients, till at least 7 mo after HSCT by high-resolution mass spectrometry, flow-cytometry, and degranulation assay. An induction of Vδ2-cell differentiation, paralleled by increased cytotoxicity of both Vδ1 and Vδ2 cells against primary leukemia blasts was associated with ZOL treatment. Cytotoxic activity was further increased in Vδ2 cells, but not in Vδ1 lymphocytes in those patients given more than one treatment. Proteomic analysis of γδ T cells purified from patients showed upregulation of proteins involved in activation processes and immune response, paralleled by downregulation of proteins involved in proliferation. Moreover, a proteomic signature was identified for each ZOL treatment. Patients given three or more ZOL infusions had a better probability of survival in comparison to those given one or two treatments (86% vs. 54%, respectively, p = 0.008). Our data indicate that ZOL infusion in pediatric recipients of αß T- and B-cell-depleted HLA-haploidentical HSCT promotes γδ T-cell differentiation and cytotoxicity and may influence the outcome of patients.

The use of a non cultured autologous cell suspension to repair chronic ulcers.

The purpose of this uncontrolled study was to evaluate the clinical effectiveness of ReCell(®) system in the treatment of chronic ulcers. From October 2011 to July 2012, 20 patients, 8 men and 12 women with a mean age of 70 years, with chronic ulcers of different aetiology that were unresponsive to conventional therapies were recruited and treated using the ReCell(®) system. Patient pain rate, scar aesthetics and patient satisfaction were assessed using a Visual Analogue Scale, Manchester Scar Scale and the Rosenberg Self-Esteem Scale, respectively. Complete ulcer healing, defined as 100% reepithelialisation was observed between 40 and 60 days in 14 patients (70%) depending on the type of ulcer and comorbidity. At day 60 postprocedure, 80% reepithelialisation was present in five patients (25%), while one patient with concomitant psoriasis had 50% reepithelialisation. Pain scores improved by day 7 postprocedure. The function and aesthetics of the ReCell-treated patients were good. It is concluded that the ReCell technique may have provided the regenerative tissue stimulation necessary for the rapid healing of chronic ulcers, including those not responsive to more traditional methods.

P.R.L. platelet rich lipotransfert: our experience and current state of art in the combined use of fat and PRP.

The authors report their experience about the use of P.R.L. PLATELET RICH LIPOTRANSFERT method (platelet rich plasma mixed fat grafting) in 223 patients affected by soft tissue defects (ulcers, Romberg syndrome, Hemifacial atrophy, loss of substance, and signs of aging). This paper introduces the reader to PRP therapy and reviews the current literature on this emerging treatment modality, showing at the current clinical use of PRP in plastic and reconstructive surgery, with description of innovative methods and future prospects. This technique provides a promising alternative to surgery by promoting safe and natural healing. Here recent studies concerning the use of PRP in the treatment of chronic ulcers and soft tissue defect are reviewed.

Treatment of stable vitiligo hands by ReCell system: a preliminary report.

BACKGROUND: The aim of this report was to analyze the results obtained with the ReCell system for the surgical treatment of stable vitiligo hands. MATERIALS AND METHODS: One patient with stable vitiligo of the hands was admitted at the Department of Plastic and Reconstructive Surgery, University of Rome Tor Vergata. The patient underwent to ReCell system for the treatment of stable vitiligo hands. RESULTS: The repigmentation was assessed using the Vitiligo Area Scoring index (VASI). The extent of pigmentation was scored as excellent, good, fair, and poor depending on the percentage of the repigmentation in the previously depigmented site. The color of the repigmented area was compared with the adjacent normally pigmented area. The patient had an excellent repigmentation. CONCLUSIONS: ReCell system is a simple, safe and feasible technique. The method that uses noncultured autologous epidermal suspension is simpler, cheaper, less time consuming and does not require sophisticated laboratory facilities, when compared with methods employing cultured melanocytes.

A case of labial fusion and urinary pseudo-incontinence in an elderly woman. A surgical treatment and a review.

Labial fusion is defined as either partial or complete adherence of the labia minora (1), and also called vulvar fusion, adhesions of the labia minor or conglutination of the labia minora and sinechia of the vulva. The complete and severe labial fusion is a rare pathology with a small number of cases reported in the literature in adults. We present a case report of a postmenopausal woman who presented with voiding difficulty and incontinence and was treated by surgical division of the adhesions and immediate resolution of the urinary incontinence confirmed by multichannel urodynamic test postoperatively.

Use of platelet rich plasma and hyaluronic acid on exposed tendons of the foot and ankle.

Platelet rich plasma was used as an autologous scaffold for cellular growth, in combination with hyaluronic acid as a temporary dermal substitute. This aided healing of acute and chronic open wounds of the foot and ankle.

Use of a novel autologous cell-harvesting device to promote epithelialization and enhance appropriate pigmentation in scar reconstruction.

BACKGROUND: Epidermal replacement is an important step in the management of patients with post-traumatic and iatrogenic scars. Skin-colour variation from disease or trauma causes significant changes in self-image and appearance. AIM: The aim of our study was to analyse the results obtained with a novel autologous cell-harvesting system (ReCell) for epidermal replacement in patients with post-traumatic scars that had not improved with any other surgical procedure. METHODS: We recruited 30 patients with post-traumatic or iatrogenic scars admitted to our department over 2 years. The primary endpoints of the study were: (i) time for complete epithelialization (both treated area and biopsy site) and (ii) aesthetic and functional quality of the epitheliaization (colour, joint contractures). Infections, inflammations or any adverse effects of the procedure were also reported. RESULTS: In total, 30 patients were analysed. The aesthetic and functional outcomes were rated by both patient and surgeon. Pigmentation was rated by the Vancouver Scar Scale. Most (80%) of the patients had an excellent or good outcome, with pigmentation rated as normal in 60% of the group. CONCLUSIONS: The procedure is a feasible, simple and safe technique. It gives similar results to skin grafting but because it harvests from much smaller areas, can open possible future applications in the management of patients with large scars.

Role of 64-slice cardiac computed tomography in the evaluation of patients with non-ST-elevation acute coronary syndrome.

PURPOSE: This study was done to evaluate the feasibility, sensitivity and specificity of 64-slice computed tomography (CT) in identifying haemodynamically significant (>50%) coronary artery stenoses in patients with suspected acute coronary syndrome (ACS) by correlating the CT findings with the clinical event and data provided by conventional coronary angiography (CCA). MATERIALS AND METHODS: Sixty-four patients (38 men and 26 women; mean age 65 years; range+/-10 years) presenting to our hospital's emergency department with a clinical suspicion of ACS were studied with 64-slice CT followed by CCA within 24 h of arrival. RESULTS: Two patients (3.1%) were excluded from the analysis due to artefacts. Per-patient analysis in the remaining 62 patients identified 24 cases (38.7%) of negative CT findings (no stenoses or stenoses <50%), 35 cases of positive CT findings (56.4%) with identification of the culprit lesion, two cases in which the culprit lesion was not identified and one patient with unconfirmed stenosis. Sensitivity and specificity were 94.6% and 96%, respectively. Per-vessel analysis (186 vessels) revealed 17 non-evaluable vessels (9.1%) due to motion artefacts, 61 vessels (32.8%) with stenosis >50%, seven overestimated vessels (3.7%) due to extensive calcifications, three vessels (1.6%) with underestimated stenosis and 98 vessels (52.6%) without stenosis. Sensitivity and specificity were 95.3% and 93.3%, respectively. CONCLUSIONS: In this type of emergency, coronary CT angiography could lead to considerably lower healthcare costs by identifying patients without coronary disease and allowing immediate discharge without any need for further diagnostic procedures.

A randomized trial comparing ReCell system of epidermal cells delivery versus classic skin grafts for the treatment of deep partial thickness burns.

BACKGROUND: Our purpose was to directly compare results obtained with the ReCell system and the classic skin grafting for epidermal replacement in deep partial thickness burns. MATERIALS AND METHODS: We recruited all patients with deep partial thickness burns admitted at the Burn Centre of S. Eugenio Hospital in Rome over 2 years. Enrollment was conducted with a controlled strategy--sampling chart--that allowed homogeneous groups (ReCell and skin grafting) for age, gender, type of burns and total burn surface area (TBSA). We evaluated as primary endpoints of the study the (i) time for complete epithelization (both treated area and biopsy site) and (ii) aesthetic and functional quality of the epithelization (color, joint contractures). Secondary endpoints were the assessment of infections, inflammations or any adverse effects of the ReCell procedure, particular medications assumed, postoperative pain. RESULTS: Eighty-two patients were analyzed in two homogeneous groups. All of them received adequate epidermal replacement, but skin grafting was faster than ReCell (p<0.05). On the contrary, ReCell biopsy areas and postoperative pain were smaller than classic grafting (p<0.05). The aesthetic and functional outcomes were similar between procedures. CONCLUSIONS: ReCell is a feasible, simple and safe technique. It gives similar results to skin grafting but, harvesting minor areas, can open possible future applications in the management of large-burns patients.

Traumas of the middle skull base with TMJ involvement. Case report.

The authors report their experience with temporomandibular joint (TMJ) traumas involving breakage of the roof of the glenoid cavity, an infrequent event that occurs in those cases in which, as a result of the condylar neck not fracturing, the traumatic energy is transmitted to the middle skull base. As the literature contains no valid series for establishing standardized protocols for the treatment of these fractures, we propose our own orthopedic-functional approach. The patient observed by us had suffered a cranio-facial trauma and presented the classical symptoms and signs of TMJ traumas and complete bilateral Bell paralysis. He was subjected to a CAT scan and then to 2-stage treatment consisting of functional rest with liquid diet followed by physiotherapy. An almost total recovery in TMJ function was observed after 1 month. At 1-year follow-up the facial paralysis had resolved completely. On the basis of our experience, breakages of the glenoid cavity can be compared, in terms of treatment procedure, to intracapsular fractures of the TMJ with surgery confined to cases of ankylosis sequelae. To avoid the onset of ankylosis careful control of clinical, functional and radiological follow-up is required.

Lack of effect of 1 year intake of a high-dose vitamin and mineral supplement on cognitive function of elderly women.

OBJECTIVE: To determine if long-term, high-vitamin supplementation could reverse cognitive malfunction in old people. METHODS: We performed a longitudinal study relating the 12-month outcome to baseline values. Twenty non-vitamin-deficient elderly females with a Folstein mini mental state examination score indicating cognitive malfunctions were recruited to ascertain if feeding a high-dose vitamin-mineral supplement for 1 year could, by mass vitamin action, reverse some existing cognitive malfunctions. Ten females were fed a high-dose vitamin-mineral supplement pill with each of three daily meals for 1 year; the other 10 did not receive this supplementation. Twelve blood vitamin analyses and a Folstein mini mental state examination were performed for each of the 20 subjects before and after 1 year; each subject served as its own control. RESULTS: No improvement in cognitive malfunction was noted despite elevation of blood vitamins. CONCLUSION: Feeding of a high-dose vitamin and mineral supplement for 1 year did not improve cognitive malfunction in non-vitamin-deficient elderly in this study.

Holotranscobalamins in B12 and non B12 requiring prokaryotes and eukaryotes.

Transcobalamins, vitamin B12 binding proteins, deliver B12 to cell surface receptors which then permit B12 to cross cell membranes for metabolic use. There is little documentation concerning B12 binding proteins in bacteria and protists. We found that prokaryotes and eukaryotes requiring B12, as well as those protists synthesizing B12, also produce several transcobalamins for functionally transporting B12 similar to humans.

HIV and dialysis: is it a real risk?

The need to expound this work was born of the necessity to confront all the organisational difficulties that present themselves with the arrival of a HIV+ patient for dialysis in our centre. On such occasions, in fact, we were unable to find a protocol, guidelines or a text that could help us in a complete and adequate way, to resolve the doubts, the fears and the practical, ethical and legal problems that we suddenly had to face.

Vitamin profile of 174 mothers and newborns at parturition.

Thiamin, biotin, B12, folate, pantothenate, riboflavin, nicotinate, B6, vitamins A, E, C, and beta-cartene was estimated in the blood of 174 mothers and in the cord blood of their neonates at parturition. A vitamin profile of normal preganancy was established for mother and neonate. This was based on values obtained from 95 percent confidence limits in 74 mothers not taking oral vitamins and 133 mothers ingesting various vitamin supllements. Circulating vitamin levels in 38 neonates born to mothers with no laboratory evidence of hypovitaminemia was persented. The higher incidence of hypobitaminemia in gravidas not taking vitamins was folate, thiamin, vitamins A, C, B12, BL and nicotinate in descending order. Ingestion of vitamins supplements reduced the incidence of hypovitaminemia. A similar relationship held for neonates from these mothers. However, despite vitamin ingestion thiamin, folate, vitamins C, A, B6, B12 and nicotinate hypovitaminemia was evident. There was an approximate 1:2-5 ratio between mother and neonate blood vitamins; vitamins A, B6, E, and beta-carotene were exceptions. Vitamin B6 ran parallel, while vitamins A, B6, E, and beta-carotene ratios reversed in favor of the mother at an approximate ratio of 2:1, 4:1 and 7:1, respectively. It is concluded that vitamin profile for mother and neonate, taking into account values established from a lorge gravid population, should now permit an evaluation of the effects of hypoviteminemia on the outcome of pregnancy and infant health.