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Abstract Background Patients undergoing heart surgery continue to be the largest demand on blood transfusions. The need for transfusion is based on the risk of complications due to poor cell oxygenation, however large transfusions are associated with increased morbidity and risk of mortality in heart surgery patients. The aim of this study was to identify preoperative and intraoperative risk factors for transfusion and create a reliable model for planning transfusion quantities in heart surgery procedures. Methods We performed an observational study on 3315 consecutive patients who underwent cardiac surgery between January 2000 and December 2007. To estimate the number of packs of red blood cells (PRBC) transfused during heart surgery, we developed a multivariate regression model with discrete coefficients by selecting dummy variables as regressors in a stepwise manner. Model performance was assessed statistically by splitting cases into training and testing sets of the same size, and clinically by investigating the clinical course details of about one quarter of the patients in whom the difference between model estimates and actual number of PRBC transfused was higher than the root mean squared error. Results Ten preoperative and intraoperative dichotomous variables were entered in the model. Approximating the regression coefficients to the nearest half unit, each dummy regressor equal to one gave a number of half PRBC. The model assigned 4 units for kidney failure requiring preoperative dialysis, 2.5 units for cardiogenic shock, 2 units for minimum hematocrit at cardiopulmonary bypass less than or equal to 20%, 1.5 units for emergency operation, 1 unit for preoperative hematocrit less than or equal to 40%, cardiopulmonary bypass time greater than 130 minutes and type of surgery different from isolated artery bypass grafting, and 0.5 units for urgent operation, age over 70 years and systemic arterial hypertension. Conclusions The regression model proved reliable for quantitative planning of number of PRBC in patients undergoing heart surgery. Besides enabling more rational resource allocation of costly blood-conservation strategies and blood bank resources, the results indicated a strong association between some essential postoperative variables and differences between the model estimate and the actual number of packs transfused.

Autochthonous hepatitis E virus (HEV) is an emerging health issue in developed countries and is thought to be a porcine zoonosis; its spread is underestimated and there is concern about the possibility of chronic infection in immunosuppressed patients; HEV transmission through blood has also been demonstrated. We conducted a retrospective study (2007-2013) on HEV seroprevalence using stored serum samples from 132 blood donors and 118 renal transplant recipients living mainly in central Italy. Anti-HEV IgG was positive in 12/132 (9.1%) of the blood donors and 12/118 (10.2%) of the transplant recipients. All subjects but one were autochthonous and none showed signs of liver disease at the time of sampling. A significant association was documented between mean age of patients and the serology against HEV especially in the group of blood donors. Our study, albeit limited and retrospective, confirms the circulation of autochthonous HEV in central Italy; the presence of antibodies against HEV in particular categories of persons such as blood donors and transplant patients, who are not screened for the infection, raises questions in terms of transfusion safety and health protection of immunocompromised patients.

Leishmaniasis is a protozoan infection endemic in Italy with a greatly underestimated prevalence. The recent documentation of parasitaemia in blood donors is a cause of concern for blood safety. Because there is no screening against leishmania, we performed a study to assess the presence of protozoa in blood donors of Siena district (Tuscany) during the seasonal activity of the vector. From June to October 2007, 162 patients were screened for Leishmania infantum by indirect immunofluorescence serology (IFAT) and PCR for kinetoplast (kDNA). No subject was positive for antibodies, while 11 samples (6.8%) were positive for kDNA. A second PCR (nested-PCR) was negative for all kDNA positive individuals and other subjects for a total of 55 samples (33% of total subjects). The sequence analysis of three samples positive for kDNA was compatible with mitochondrial DNA. Through the techniques used, we were unable to confirm the presence of leishmania in the blood of the subjects studied. The choice of the diagnostic protocol in blood donors remains an open issue as molecular analysis (kDNA) seems to suggest, in our experience, limits of specificity.

Background: Patients undergoing heart surgery continue to be the largest demand on blood transfusions. The need for transfusion is based on the risk of complications due to poor cell oxygenation, however large transfusions are associated with increased morbidity and risk of mortality in heart surgery patients. The aim of this study was to identify preoperative and intraoperative risk factors for transfusion and create a reliable model for planning transfusion quantities in heart surgery procedures., Methods: We performed an observational study on 3315 consecutive patients who underwent cardiac surgery between January 2000 and December 2007. To estimate the number of packs of red blood cells (PRBC) transfused during heart surgery, we developed a multivariate regression model with discrete coefficients by selecting dummy variables as regressors in a stepwise manner. Model performance was assessed statistically by splitting cases into training and testing sets of the same size, and clinically by investigating the clinical course details of about one quarter of the patients in whom the difference between model estimates and actual number of PRBC transfused was higher than the root mean squared error., Results: Ten preoperative and intraoperative dichotomous variables were entered in the model. Approximating the regression coefficients to the nearest half unit, each dummy regressor equal to one gave a number of half PRBC. The model assigned 4 units for kidney failure requiring preoperative dialysis, 2.5 units for cardiogenic shock, 2 units for minimum hematocrit at cardiopulmonary bypass less than or equal to 20%, 1.5 units for emergency operation, 1 unit for preoperative hematocrit less than or equal to 40%, cardiopulmonary bypass time greater than 130 minutes and type of surgery different from isolated artery bypass grafting, and 0.5 units for urgent operation, age over 70 years and systemic arterial hypertension., Conclusions: The regression model proved reliable for quantitative planning of number of PRBC in patients undergoing heart surgery. Besides enabling more rational resource allocation of costly blood-conservation strategies and blood bank resources, the results indicated a strong association between some essential postoperative variables and differences between the model estimate and the actual number of packs transfused.

The aim of the present study was to demonstrate the mitogenic and differentiating properties of platelet-rich plasma releasates (PRPr) on human chondrocytes in mono- and three-dimensional cultures. In order to assess if PRPr supplementation could maintain the chondrocyte phenotype or at least inhibit the cell de-differentiation even after several days in culture, we performed a proteomic study on several cell cultures independently grown, for different periods of time, in culture medium with FCS, human serum (HS), and releasates obtained from PRP and platelet-poor plasma (PPP). We found that PRP treatment actually induced in chondrocytes the expression of proteins (some of which novel) involved in differentiation.

Rotator cuff tears are common soft-tissue injuries that often require surgical treatment. Initial efforts to better tendon healing centered on improving the strength of repair. More recent studies have focused on abiologic enhancement of the healing process. Platelet rich plasma (PRP) is a fraction of plasma that has been isolated and used to enhance regeneration in bone and soft tissues. The healing potential of PRP has been attributed to the release of multiple growth factors from the highly concentrated platelets. The "Cascade" membrane is a thin layer of autologous fibrine that is very rich in platelets and is obtained by high speed centrifugation of a small quantity of PRP. The Authors present the case of C.U., a right-handed 53 year-old male that came to our attention complaining of severe right shoulder pain and ROM reduction. The MRI showed a thick tear of the sovraspinatus tendon with retraction of the muscle. The patient underwent surgical repair with arthroscopy and mini-open approach with acromionplasty, subacromial decompression, cuff repair by trans-bone suture and application of the "Cascade" membrane (A.T. Grade Milano, Italy) that was sutured side-to-side to close the 10 mm tear. A standard rehabilitation protocol for rotator cuff tears was performed. A MRI 6 months after surgical repair showed the complete integrity of the cuff under the membrane. Currently no widely accepted treatment for massive, irreparable rotator cuff tears is available. Allograft reconstruction to span the remaining defect in massive rotator cuff tears will lead to increase functional results; we believe that further studies are needed to describe and evaluate its potential in tendon healing; multiple MRI studies will demonstrate healing of the tendon with the graft.

The potential therapeutic value and versatility of platelet-derived products has recently stimulated the research and interest in the field of regenerative medicine. Platelet gels (PG), generated by thrombin-activated platelets, represent a new biotechnology for stimulation and acceleration of tissue healing and regeneration. However, despite the diffused and successful use of PG in clinical practice, a more detailed knowledge of the cellular and molecular mechanisms involved is required. In the present study, we show that human peripheral blood mononuclear cells (PBMC) co-cultured with PG, in the presence of the inflammatory activator lipopolysaccharide, secreted higher amounts of pro-inflammatory and pro-angiogenic cytokines, such as interleukin (IL)-1beta, IL-6 and IL-8. In contrast, the release of the anti-angiogenic cytokines interferon-gamma and IL-12 was significantly reduced. In addition the production of the anti-inflammatory cytokine IL-10 was not affected by PG. Finally, hypoxia, a common feature of the healing tissue, potentiated the effects exerted by PG on the release of IL-1beta by PBMC. In conclusion, PG treatment reveals a unique capacity of articulating a pro-inflammatory and pro-angiogenic cytokine profile in human PBMC, which may partially explain the clinical success of PG application in a wide range of diseases.

Background: Several experimental models have shown that CXCL10 is required for initiation and development of graft failure caused by both acute and chronic rejection., Methods: CXCL10 expression and distribution was investigated in tissue specimens obtained from 22 patients suffering from acute rejection (AR) or chronic allograft nephropathy (CAN) by using in situ hybridization. Furthermore, pretransplantation sera of 316 cadaveric kidney-graft recipients were tested retrospectively for serum CXCL10 levels by a quantitative sandwich immunoassay., Results: Bioptic specimens obtained from patients with CAN were characterized by wide CXCL10 expression not only at level of infiltrating inflammatory cells but also of vascular, tubular, and glomerular structures. In addition, assessment of pretransplant serum CXCL10 levels in 316 graft recipients and stratification of patients in three groups according to serum CXCL10 levels (<100 pg/mL, n=163; 100-150 pg/mL, n=69; >150 pg/mL, n=84) showed highly significant differences in 5-year survival rates for the two extreme groups (95.7% vs. 79.7%, P=0.0002). Accordingly, patients who developed severe, early AR (277.14+/-65.08 p=0.004) and those who developed CAN also showed increased pretransplant serum CXCL10 levels (193.2+/-36.9, P=0.03). Multivariate analysis demonstrated that among the analyzed variables, CXCL10 (relative risk [RR] 2.801) and delayed graft function (RR 3.728) had the highest predictive power of graft loss., Conclusions: These results suggest that pretransplant serum CXCL10 levels greater than 150 pg/mL confer an increased risk of early, severe, AR and subsequent CAN, finally resulting in renal-allograft failure. This finding might be used for the individualization of immunosuppressive therapies.

In experimental models, the chemokine CXCL10/IP-10 is required for graft failure owing to both acute and chronic rejection. In the present study, pretransplantation sera from 316 cadaver kidney graft recipients were tested for serum CXCL10 and CCL22/MDC levels by an ELISA assay. Kidney graft recipients with normally functioning grafts showed significantly lower serum CXCL10 levels than patients who experienced graft failure, whereas no differences for serum CCL22 levels were observed. After the assignment of all patients to four groups according to serum CXCL10 levels, the death-censored survival rates of grafts were 97.5%, 93.6%, 89.7%, 78.7% (p = 0.0006) at 5 years, while no influence was observed on patient survival. Accordingly, patients with the highest CXCL10 levels showed an increased frequency and severity of rejection episodes. Serum C-reactive protein (CRP) level was also assayed in the same samples. Increase of serum CRP levels represented a predictive parameter for death, but not for graft failure. Multivariate analysis demonstrated that among the analyzed variables, CXCL10 had the highest predictive power of graft loss (RR 2.787). Thus, measurement of pretransplant serum CXCL10 levels might represent a clinically useful parameter to identify subjects who are at high risk of severe rejection and graft failure., (Copyright 2004 Blackwell Munksgaard)