Bispecific antibodies targeting tumor-associated antigens and neutralizing complement regulators increase the efficacy of antibody-based immunotherapy in mice.

The efficacy of antibody-based immunotherapy is due to the activation of apoptosis, the engagement of antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity (CDC). We developed a novel strategy to enhance CDC using bispecific antibodies (bsAbs) that neutralize the C-regulators CD55 and CD59 to enhance C-mediated functions. Two bsAbs (MB20/55 and MB20/59) were designed to recognize CD20 on one side. The other side neutralizes CD55 or CD59. Analysis of CDC revealed that bsAbs could kill 4-25 times more cells than anti-CD20 recombinant antibody in cell lines or cells isolated from patients with chronic lymphocytic leukemia. The pharmacokinetics of the bsAbs was evaluated in a human-SCID model of Burkitt lymphoma. The distribution profile of bsAbs mimics the data obtained by studying the pharmacokinetics of anti-CD20 antibodies, showing a peak in the tumor mass 3-4 days after injection. The treatment with bsAbs completely prevented the development of human/SCID lymphoma. The tumor growth was blocked by the activation of the C cascade and by the recruitment of macrophages, polymorphonuclear and natural killer cells. This strategy can easily be applied to the other anti-tumor C-fixing antibodies currently used in the clinic or tested in preclinical studies using the same vector with the appropriate modifications.

Acute pyelonephritis as a cause of hyponatremia/hyperkalemia in young infants with urinary tract malformations.

Obstructive uropathy causes tubular resistance to aldosterone and severe metabolic imbalance may be precipitated by an episode of pyelonephritis. In the last 3 years we investigated 52 episodes of pyelonephritis (positive urine culture, elevated C reactive protein, fever, elevated neutrophil count) in 50 children between 15 days and 15 months of age. Ultrasonography voiding cystography and renal scintiscan were performed in all cases and i.v. urography in some. A salt-losing syndrome with hyponatremia and hyperkalemia (Na < 125 meq/liter; K > 6.3 meq/liter) was observed in 17 infants < 3 months, accompanied by plasma aldosterone concentration of 5000 to 23,000 pg/ml (normal value, < 1000 pg/ml). All these children had a severe urinary tract (UT) malformation (ureteropelvic junction stenosis in 7 cases, vesicoureteral reflux in 7, posterior urethral valves in 2, double system in 1). Thirteen infants < 3 months, 7 with no urinary tract malformations, did not have electrolyte imbalance. Pyelonephritis was diagnosed in 20 other patients ages 4 to 15 months, including 16 with severe UT malformations; 4 had normal UTs. We conclude that a salt-losing syndrome with tubular resistance to aldosterone can occur during pyelonephritis in young infants with congenital UT malformation, that the risk diminishes considerably or disappears after 3 months of age and that in the absence of UT malformation pyelonephritis does not cause acute sodium loss of clinical relevance.

[Diagnostic and therapeutic approach in uropathic abnormalities diagnosed in utero]. / Approccio diagnostico e terapeutico delle uropatie malformative diagnosticate in utero.

Ultrasonography permits to suspect urinary tract abnormalities in fetal age: hydronephrosis is one of the most frequent anomalies detected, but renal dysplasia (multicystic kidney, renal agenesis, renal cysts) can be found in some cases. At birth, if abnormalities are confirmed by ultrasound scan other instrumental investigation are needed: cystography, to exclude vesicoureteric reflux, and sequential scintigraphy, to evaluate if obstruction of urinary tract is present or to confirm the suspect of renal agenesis or multicystic dysplasia. Urography could be necessary in some cases. For bilateral uropathy evaluation of renal function with blood tests is required. Early surgery is needed in case of severe obstructive uropathy; if obstruction is mild the patients can be followed with instrumental investigation. Antibiotic prophylaxis is required to prevent urinary tract infections in patients with vesicoureteric reflux. The indications for surgery in patients with vesico-ureteric reflux are still unclear.

Biosensing and environmental sensing for emergency and protection e-Textiles.

The ProeTEX project introduced for the first time a complete set of smart garments integrating sensors for the physiological and environmental monitoring of emergency operators. These "smart" garments have been deeply tested in emergency-like contexts by professional rescuers, in order to assess real-time acquisition, processing and transmission of data from moving subjects while operating in harsh conditions. Here we report an overview of the main results obtained during field trials performed in 2010 by Italian and French professional firefighters, in specialized training centers, while dressing the ProeTEX prototypes. Results clearly demonstrate the benefit and step forward of such a system in order to monitor and coordinate rescuers even during intervention far away from the emergency headquarter.

Heart rate and accelerometer data fusion for activity assessment of rescuers during emergency interventions.

The current state of the art in wearable electronics is the integration of very small devices into textile fabrics, the so-called ¿smart garment.¿ The ProeTEX project is one of many initiatives dedicated to the development of smart garments specifically designed for people who risk their lives in the line of duty such as fire fighters and Civil Protection rescuers. These garments have integrated multipurpose sensors that monitor their activities while in action. To this aim, we have developed an algorithm that combines both features extracted from the signal of a triaxial accelerometer and one ECG lead. Microprocessors integrated in the garments detect the signal magnitude area of inertial acceleration, step frequency, trunk inclination, heart rate (HR), and HR trend in real time. Given these inputs, a classifier assigns these signals to nine classes differentiating between certain physical activities (walking, running, moving on site), intensities (intense, mild, or at rest) and postures (lying down, standing up). Specific classes will be identified as dangerous to the rescuer during operation, such as, ¿subject motionless lying down¿ or ¿subject resting with abnormal HR.¿ Laboratory tests were carried out on seven healthy adult subjects with the collection of over 4.5 h of data. The results were very positive, achieving an overall classification accuracy of 88.8%.

Bio-inspired controller for a dexterous prosthetic hand based on Principal Components Analysis.

Controlling a dexterous myoelectric prosthetic hand with many degrees of freedom (DoFs) could be a very demanding task, which requires the amputee for high concentration and ability in modulating many different muscular contraction signals. In this work a new approach to multi-DoF control is proposed, which makes use of Principal Component Analysis (PCA) to reduce the DoFs space dimensionality and allow to drive a 15 DoFs hand by means of a 2 DoFs signal. This approach has been tested and properly adapted to work onto the underactuated robotic hand named CyberHand, using mouse cursor coordinates as input signals and a principal components (PCs) matrix taken from the literature. First trials show the feasibility of performing grasps using this method. Further tests with real EMG signals are foreseen.

P-selectin glycoprotein ligand-1 as a potential target for humoral immunotherapy of multiple myeloma.

Monoclonal antibodies (mAbs), successfully adopted in the treatment of several haematological malignancies, have proved almost ineffective in multiple myeloma (MM), because of the lack of an appropriate antigen for targeting and killing MM cells. Here, we demonstrate that PSGL1, the major ligand of P-Selectin, a marker of plasmacytic differentiation expressed at high levels on normal and neoplastic plasma cells, may represent a novel target for mAb-mediated MM immunotherapy. The primary effectors of mAb-induced cell-death, complement-mediated lysis (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC), were investigated using U266B1 and LP1 cell-lines as models. Along with immunological mechanisms, the induction of apoptosis by PSGL1 cross-linking was assessed. The anti-PSGL1 murine mAb KPL1 induced death of MM cells in a dose- and time-dependent fashion and mediated a significant amount of ADCC. KPL1 alone mediated C1q deposition on target cells but proved unable to induce CDC due to inhibition of the lytic activity of complement by membrane complement regulators (mCRP) expressed on the cell surface. Consistently, CDC was induced by KPL1 upon mCRP blockage. Our results suggest a role for PSGL1 in MM humoral immunotherapy and support further in vivo studies assessing the effects of anti-PSGL1 mAbs on MM growth and interaction with the bone marrow microenvironment.

Congenital reflux nephropathy: a follow-up of 108 cases diagnosed perinatally.

OBJECTIVE: To determine: (i) the proportion of vesicoureteric reflux (VUR) associated with congenital renal damage and whether it can be severe enough to cause renal impairment from birth: (ii) to evaluate the distribution of males and females affected; and (iii) to describe the course of congenital damage in the first years of life. PATIENTS AND METHODS: A total of 108 children (76 male and 32 female, M:F 2.3:1), whose VUR was diagnosed before any infection, were followed from birth for a mean (range) of 4.3 (1-10) years. Renal damage was defined by serum creatinine concentration, creatinine clearance and renal imaging (ultrasonography and renal scintigraphy) performed within the first month of life and periodically thereafter. RESULTS: Of the 108 children, 58 had bilateral and 50 unilateral reflux (total number of refluxing units, 166). High-grade VUR (grade > or = 4) was found in 96 (58%) refluxing renal units (RRUs). Males had a prevalence of bilateral severe (> or = grade 4) reflux (M:F 5.2:1), while in those wit unilateral VUR, the M:F ration was 1.5:1. At birth, mild to moderate damage was present in 56 (36%) RRUs and only associated with VUR of grade > or = 3. Bilateral reflux of grade > or = 4 was associated with congenital moderate/severe renal failure in nine neonates (seven males). In infants with grade > or = 4 VUR who underwent surgical correction, VUR resolved in 92% of cases. In infants with VUR of grade > or = 4 followed medically, the reflux spontaneously resolved in 42% and ameliorated in 16% after 18 months. Serial renal scans during the follow-up showed no progression of renal damage. CONCLUSIONS: VUR diagnosed at birth on prenatal ultrasonography is associated with congenital damage, with males affected more often than females. The damage involves both kidneys in a consistent proportion and is an important cause of chronic renal impairment from birth. It does not progress in the first years of life if infections are prevented. It is suggested that males with this condition may constitute a major group at risk of developing chronic renal failure in later life.