Anna Maria Patti

Experimental posterolateral spinal fusion with porous ceramics and mesenchymal stem cells

Alternatives to autogenous bone graft for spinal fusion have been investigated for many years. It has been shown that osteoconductive materials alone do not give a rate of fusion which is comparable to that of autogenous bone graft. We analysed the effectiveness of porous ceramic loaded with cultured mesenchymal stem cells as a new graft material for spinal fusion in an animal model. Posterolateral fusion was carried out at the L4/L5 level in 40 White New Zealand rabbits using one of the following graft materials: porous ceramic granules plus cultured mesenchymal stem cells (group I); ceramic granules plus fresh autogenous bone marrow (group II); ceramic granules alone (group II); and autogenous bone graft (group IV). The animals were killed eight weeks after surgery and the spines were evaluated radiographically, by a manual palpation test and by histological analysis. The rate of fusion was significantly higher in group I compared with group III and higher, but not significantly, in group I compared with groups II and IV. In group I histological analysis showed newly formed bone in contact with the implanted granules and highly cellular bone marrow between the newly formed trabecular bone. In group II, thin trabeculae of newly formed bone were present in the peripheral portion of the fusion mass. In group III, there was a reduced amount of newly formed bone and abundant fibrous tissue. In group IV, there were thin trabeculae of newly formed bone close to the decorticated transverse processes and dead trabecular bone in the central portion of the fusion mass. In vitro cultured mesenchymal stem cells may be loaded into porous ceramic to make a graft material for spinal fusion which appears to be more effective than porous ceramic alone. Further studies are needed to investigate the medium- to long-term results of this procedure, its feasibility in the clinical setting and the most appropriate carrier for mesenchymal stem cells.

Molecular techniques for the identification of enteric viruses in marine waters

Abstract The RNAs extracted from 28 high spin pellets of clarified lysates from BGM cells, previously infected by concentrated Adriatic Sea water samples, were analyzed by a dot-blot test. We used a 32 P-labeled cDNA probe ( BamH I 220–670 ) from the 5′ non-coding end of the cloned poliovirus I (Mahoney) genome. The probe was selected for its broad range of genetic specificities. Based on the density of the hybridization signals on the autoradiograph, our dot-blot results showed a high degree of reactivity of the probe to homologous (poliovirus 1) and heterologous (echovirus 6 and coxsackievirus B3) reference RNA, as predicted by a computer analysis of their sequences, and a low reactivity to that of the samples leading us to exclude the presence of enteroviruses like the reference strains. In order to understand if there were other enteroviral serotypes or not, dot-blot tests were supplemented with Northern-blot hybridization assays. Results from the Northern-blot showed a series of fragments ranging from 4.3 to 1.2 kb but no signal corresponding to 7.5 kb (as did the positive control RNAs). These features suggested that the tested RNAs might derive from viruses of the Reoviridae family, which includes members with segmented genome. Traditional PAGE analysis showed clearly the 10-fragments pattern characteristic of reoviruses. Twenty-three out of the twenty-eight samples tested showed the presence of viruses, and this confirms the previously noted cytopathic effect (CPE) of the samples on the BGM cells.

Sporadic isolation of sabin-like polioviruses and high-level detection of non-polio enteroviruses during sewage surveillance in seven Italian cities, after several years of inactivated poliovirus vaccination.

ABSTRACT Sewage surveillance in seven Italian cities between 2005 and 2008, after the introduction of inactivated poliovirus vaccination (IPV) in 2002, showed rare polioviruses, none that were wild-type or circulating vaccine-derived poliovirus (cVDPV), and many other enteroviruses among 1,392 samples analyzed. Two of five polioviruses (PV) detected were Sabin-like PV2 and three PV3, based on enzyme-linked immunosorbent assay (ELISA) and PCR results. Neurovirulence-related mutations were found in the 5′ noncoding region (5′NCR) of all strains and, for a PV2, also in VP1 region 143 (Ile > Thr). Intertypic recombination in the 3D region was detected in a second PV2 (Sabin 2/Sabin 1) and a PV3 (Sabin 3/Sabin 2). The low mutation rate in VP1 for all PVs suggests limited interhuman virus passages, consistent with efficient polio immunization in Italy. Nonetheless, these findings highlight the risk of wild or Sabin poliovirus reintroduction from abroad. Non-polio enteroviruses (NPEVs) were detected, 448 of which were coxsackievirus B (CVB) and 294 of which were echoviruses (Echo). Fifty-six NPEVs failing serological typing were characterized by sequencing the VP1 region (nucleotides [nt] 2628 to 2976). A total of 448 CVB and 294 Echo strains were identified; among those strains, CVB2, CVB5, and Echo 11 predominated. Environmental CVB5 and CVB2 strains from this study showed high sequence identity with GenBank global strains. The high similarity between environmental NPEVs and clinical strains from the same areas of Italy and the same periods indicates that environmental strains reflect the viruses circulating in the population and highlights the potential risk of inefficient wastewater treatments. This study confirmed that sewage surveillance can be more sensitive than acute flaccid paralysis (AFP) surveillance in monitoring silent poliovirus circulation in the population as well as the suitability of molecular approaches to enterovirus typing.

Anti-Purkinje cell antibody as a biological marker in attention deficit/hyperactivity disorder: a pilot study.

An autoimmune hypothesis has been suggested for several disorders in childhood. The aim of the study was to clarify the role of the cerebellum in ADHD and to evaluate the possible association between anti-Yo antibodies and ADHD. The presence/absence of antibodies was tested by indirect immunofluorescence assay on 30 combined subtype ADHD children, on 19 children with other psychiatric disorders (Oppositional-defiant and Conduct Disorders, Dyslexia) and 27 healthy controls. Results showed a significant positive response to the anti-Yo antibody immunoreactivity in the Purkinje cells of the cerebellum of ADHD children, compared with the control group and the psychiatric non-ADHD children. This association points to an immune dysregulation and the involvement of the cerebellum in ADHD.

Culture of human rotator cuff cells on orthobiologic support (porcine small intestinal submucosa

Outcomes obtained in patients with two-tendon rotator cuff tear submitted to repair reinforced with porcine small intestinal submucosa (SIS) have not been as encouraging as those observed in animal models. We verify the capacity of SIS to be used as a physical support for a culture of cuff cells. During arthroscopic repairs of large rotator cuff tears, we removed a fragment of supraspinatus tendon. Samples were treated for obtaining a cuff cell culture. Daily microscopic analysis, to observe adhesion to substrate, replication and cell shape was performed. A confluent monolayer was obtained in 1 week. Cells at the second passage were collected and seeded onto scaffold and cultured for 7–30 days. A morphological and immunohistochemical evaluation was performed. After 1 week, a monolayer of tendinous-like cells lay along the surface of the SIS. Within two weeks, a multicellular layer was observable in many foci of the scaffold. After a month, the cells completely invaded the numerous splits of the SIS and were positive to monoclonal anti-type I collagen antibody. Our experimental study has proved that a cuff cell culture can be performed using SIS as substrate. The culture covers the SIS surface, therefore it may reduce immune or non-specific inflammatory reactions.

Borna disease virus infection in Italian children. A potential risk for the developing brain

Borna disease virus (BDV) infection occurs worldwide in several vertebrate species, including humans. Sero-epidemiological studies have consistently shown an increased BDV seroprevalence in neuropsychiatric patients (1–8). Because of the possible association with psychiatric diseases such as depression, research is presently conducted worldwide on this neurotropic virus. The greater part of the investigators has focused the attention on patients with psychiatric disorders, and the healthy subjects have been studied only as control group. Because extensive epidemiological studies in the general population are lacking, it is difficult to have a clear picture of the human spread of this virus, of the age of the first infection, and of the mode of transmission of this virus, including vectors or reservoirs . In addition, it is still difficult to indicate which population groups are most at risk and the natural history of the human infection (9). Even if a small fraction of psychiatric diseases could be attributed to viruses, this would be very important as viral infections can be preventable. Even though in Italy infection in horses is known and has recently been studied with new methods (10), no data are available on the role of this emerging pathogen among humans. The goal of this work was to monitor the prevalence of BDV infection among Italian children. BDV infection in childhood was unexplored worldwide except for unpublished information by Shollbach and Bode, presented in

Borna Disease Virus infection in children with psychiatric disorders

INTRODUCTIONIn the past 15 years the scientific communityhas observed a progressive change of paradigmabout psychiatric illness in adults and inchildren. Overall in the studies about mentaldiseases in children, Great emphasis was givento the mother-child relationship and to the fam-ily environment. Many studies have now sup-ported a change from a psychological view ofmental disorders to a more biological interpre-tation of psychiatry. The major contributionwas given by several studies about the geneticpredisposition of mental diseases, such asMajor Depression, Bipolar Disorder, ObsessiveCompulsive Disorder, and Attention Deficit/Hyperactive Disorder (ADHD). One of themost interesting hypothesis was suggesting thatbehavioural changes are the consequence ofviral infections. Herpes simplex virus seems tohave some relevance in the onset of schizo-phrenia (1), with a special affinity for poly-morphisms in MICB (2). These results providean intriguing hypothesis about a combinationof viral and genetic factors. Moreover severalreviews provide detailed information about Bor-na Disease Virus (BDV) infections in humans(3–5), which preferentially infect the nervoussystem being involved in neurological diseasesand in abnormal mental status. Data reported,ranged from high frequencies of BDV infectionto no BDV RNA in psychiatric patients, andfrom no significant level of BDV structural ele-