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.

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-atherogenic effects of 17ß-estradiol.

Estrogens are secreted primarily by the ovaries and placenta, by the testes in men and also produced by peripheral steroidogenic conversion. The 3 major naturally occurring estrogens are: 17ß-estradiol (E2), estrone and estriol, of which E2 is the predominant and most active. The actions of E2 are mediated by at least 3 different receptors - the classical ERs (ERα and ERß) and G-protein coupled receptor 30 (GPR30). E2 signaling in cardiomyocytes involves ERα- and ERß-independent pathways, and treatment with the E2 receptor antagonists (Selective Estrogen Receptor Modulators- SERMs), which are agonists of GPR30, inhibits cardiac cell growth. Effects of E2 in preventing endothelial dysfunction, a prerequisite of atherosclerosis, are well recognized. Atherosclerosis involves interaction between the cells of the arterial wall endothelial cells (EC) and vascular smooth muscle cell (VSMC), as well as migration of macrophages into wall tunica media. It is predominantly developed at sites with abnormally high shear stress, such as bifurcations or branching of arteries, initiated by an injury to the endothelium and exposure to atherogenic lipids and toxins, such as those contained in tobacco smoke or infectious agents. Animal studies have shown effects of E2 in preventing atherosclerosis, inflammation and endothelial or vascular dysfunction. Gender differences along this pathogenic pathway have been also described. We review the data from the available animal and human studies, which focus on anti-atherogenic effects of E2. These studies represent evidence, albeit indirect, for an inhibitory effect of E2 on the progression of coronary artery atherosclerosis.

Prediction of vascular events in subjects with subclinical atherosclerosis and the metabolic syndrome: the role of markers of inflammation.

AIM: The presence of the metabolic syndrome (MS) increases cardiovascular morbidity and mortality and we aimed to assess the outcome in subjects with the MS and subclinical atherosclerosis. METHODS: We followed-up for five years 339 Mediterranean subjects with asymptomatic carotid intima-media thickness >0.9 mm (men: 60%; age: 66±5 years), of whom 130 had the MS (men: 59%; age: 66±5 years), evaluating at baseline traditional cardiovascular risk factors (including male gender, older age, obesity, hypertension, diabetes, smoking, family history of cardiovascular diseases, dyslipidemia) and plasma levels of C-reactive protein and fibrinogen. RESULTS: Cardio- and cerebrovascular events were registered in the 29% of subjects with the MS and in the 20% of those without it and the presence of more criteria for the diagnosis of the MS was significantly associated with vascular morbidity and mortality. By multivariate analysis, including all baseline variables, independent predictive roles for the events were found for elevated markers of inflammation (OR 3.8), elevated fasting glucose (OR 2.1) and elevated triglycerides (OR 1.4). CONCLUSION: These findings confirm a worst vascular outcome in subjects with more criteria for the diagnosis of the MS and further suggest the need of future research to understand the combined role of inflammation and the MS in the progression from subclinical to clinical atherosclerosis.

[From the goal of the global eradication of polio to the recent epidemics: reflections on the present epidemiological situation and future perspectives]. / Dall'obiettivo di eradicazione mondiale della polio alle recenti epidemie: riflessioni sulla situazione epidemiologica nazionale e le sue prospettive.

The recent epidemic of polio caused by the Wilde Virus which during last spring seriously hit Tagikistan, has now caused a crisis in the polio free situation of the European Region of the OMS. In addition, considering that there has been a sudden epidemic in Congo with a shift of the age of the first infection towards adolescents/adults and an unexpected death-to-case rate, it is worth reconsidering the risk of importing the Wild Virus into countries already polio-free. In consideration of the fact that Italy is a Country at high risk for importing polioviruses we analyzed the literature of the immunologic coverage of the virus in Italy, in order to verify the rationale for the administration of a fifth dose of polio vaccine to adolescents, what has already been introduced into the vaccination calendar in other European Countries such as United Kingdom, France and Germany.

Cytomegalovirus (AD169) infection in human chondrocyte cell cultures.

Human chondrocytes are susceptible to productive human cytomegalovirus (HCMV, AD169 strain) infection. Infection of chondrocytes resulted in pronounced cytopathic effects including cell rounding and aggregation, fusions, and lysis. Immunohistochemical staining reveals that a portion of chondrocytes expressed antigens reactive to monoclonal antibodies against early HCMV proteins. The interaction of HCMV with cartilage cells may be one key for elucidating the etiopathogenesis of articular diseases.

HEV prevalence in the general population and among workers at zoonotic risk in Latium Region.

Hepatitis E Virus (HEV) infections are responsible for large waterborne outbreaks in developing countries. Sporadic cases in the developed world are mainly imported from endemic areas. HEV has been suggested to be a zoonotic infection, for which pigs may be the reservoir; specific swine strains of HEV have been identified. Humans are susceptible to infections with swine strains. The aim of this study was to analyse whether Italian pig farmers, veterinarians and abattoir workers are more exposed than persons with other occupations. A total of 92 workers at zoonotic risk and 3511 controls from the general population of two Latium cities, Rome and Rieti, were tested for IgG-HEV antibodies. No significant difference in anti-HEV prevalence was observed between the two groups. The prevalence of general population was 2.9% against 3.3% of pig breeders, while there was a statistically significant difference (p = .0004) between subjects recruited in Rome (prevalence 2.5%) and those recruited in Rieti (prevalence 5.5%). Moreover, in some subgroups of general population and in a subgroup of pig breeders, the prevalence was higher than that previously reported in Italy and in other European countries. The highest value (33%) was found in male housekeepers enrolled in Rome; an analogous value was found in the employees of abattoirs (33%). Further studies are needed to elucidate the transmission routes.

[Osteoporosis: problems and considerations of economical nature]. / Osteoporosi: problematiche e considerazioni di carattere economico.

In 1991 WHO classified osteoporosis as a major social disease, on the basis of its high prevalence, expected to rise in the future, its physical and psychological consequences and its economical costs for both the society and the individual. In the past, costs, especially the undirected and intangible ones, have been largely underestimated, due to their complex quantification. A more accurate and a deeper evaluation of the pathology and its economical burden on National Health Services will lead to a better planning of prevention strategies.

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.

Environmental surveillance of poliovirus in Italy: pilot study.

A pilot study on environmental surveillance of poliovirus included one hundred ninety-six samples (drinking water, surface water, seawater and sewage), collected between 1996 and 1998 in different regions of Italy. Samples were screened for the presence of poliovirus and other enteric viruses. Twelve polioviruses, 35 non polio enteroviruses (NPE) and 51 enteric non entero (NE) viruses were isolated. All poliovirus isolates, namely four type 1, four type 2 and four type 3, were characterised as Sabin-like strains. Thirty-one Coxsackie B and 4 Echo viruses were also detected. The presence of Sabin-like polioviruses in the environment is not unexpected since immunization with only oral poliovirus vaccine was performed in Italy until May 1999, when a sequential schedule consisting of two doses of inactivated polio vaccine (IPV) and two of oral polio vaccine (OPV) was adopted; in July 2002 a schedule consisting of four doses of IPV was adopted. Although this is only a pilot study and the number of samples analysed is limited, so far data from this study so far supports once again evidence of lack of circulating wild poliovirus and is in line with results from AFP surveillance in Italy and stool survey on healthy children.

[Viral zoonoses]. / Le zoonosi da virus.

In different areas of the World, whether in industrialized or developing countries, zoonoses represent an emerging Public Health concern either as known agents appear in areas or species in which they had not been reported or as new pathogens parasite Human as consequence of successful host switching. Epidemiological data outlined that vectorborne viral zoonoses are of particular relevance. This paper review the changes in epidemiology of bat rabies and arbovirus encephalitis.

Effect of hyaluronic acid on human chondrocyte cell lines from articular cartilage.

The effects of hyaluronic acid (HA) derivative on the proliferation and metabolism of human chondrocytes were examined. Cells were obtained from cartilage from metatarsal phalangeal joints of 20 adult humans (aged 22-63) and from femoral knee condyles of 10 subjects (aged 22-77). Chondrocytes isolated by collagenase/Dnase digestion were cultured with addition of different doses of HA for 4 weeks. Morphological studies demonstrated that HA enhanced the adhesion of cells to substrate; HA-treated chondrocytes proliferated better than chondrocytes cultured in HA-free medium. This study shows that HA improves in vitro substrate adhesion ability and proliferative activity of human cartilage cells and that the response to the treatment varies on an individual basis.

Enterovirus surveillance of Italian healthy children.

Surveillance of acute flaccid paralysis (AFP) is the golden strategy recommended by the WHO to verify the condition of polio eradication in a country. Because of the difficulty to detect all of the expected AFP cases and to reach the target incidence of 1/100,000 requested by WHO, the surveillance of enteroviruses in the population has been adopted by several countries as an important additional method to verify the absence of wild-poliovirus circulation. To complete the results of AFP surveillance set up in Italy in 1996, we have conducted a wild poliovirus surveillance by examining stool samples from 1551 healthy children aged less than 5 years, collected during the period January 1997 to January 1998. The children were from three cities (Parma, Rome and Bari) located in northern, central and southern Italy. Thirty-nine polioviruses, 72 non-polio enteroviruses and 50 enteric, nonentero (NE), viruses were isolated from stool specimens. Polioviruses identified were nine type 1, seven type 2 and twenty-three type 3. Characterization of isolates by both antigenic and molecular methods showed that all polioviruses were of vaccine origin. As expected, most polioviruses, especially types 2 and 3, presented retromutations known to be associated with loss of the Sabin attenuated phenotype. The results of this study support the data obtained from the active AFP surveillance conducted in Italy in the same period--on the absence of paralytic disease due to wild poliovirus--and altogether demonstrate the effectiveness of the vaccination program.

[Flaccid paralysis surveillance in the Latium Region]. / La sorveglianza della paralisi flaccida nella Regione Lazio.

The goal of World Health Organization is to reach the global eradication of poliomyelitis during the first decade of the third millennium. To achieve the certification of the eradication of the disease the main strategy is the Acute Flaccid Paralysis (AFP) surveillance. In Italy the active AFP surveillance was performed at national level since 1997. In the Latium region the active surveillance was performed since January 1997 by the laboratory of virology of Institute of Hygiene G Sanarelli which established a regional hospital network. During the years of survey 7 cases were found in 1997 (0.87/100,000), 4 in 1998 (0.5/100,000), 2 in 1999 (0.25/100,000) and 2 in 2000. No wild polioviruses were detected.

Human chondrocyte cell lines from articular cartilage of metatarsal phalangeal joints.

Chondrocytes can be isolated from human adult cartilage from metatarsal phalangeal joints. After enzymatic digestion to isolate viable cells, confluent monolayers were obtained 2-4 weeks after the start of cell division. Chondrocytes cultures, initiated and maintained in HAM's F12 with bovine fetal serum without the addition of other growth factors, produced in vitro a matrix rich in collagen and proteoglycans. Although several studies reported phenotypic instability, our results showed that the cell retain for more than 5 months in culture their differentiated characteristics, including the ability to produce cartilage-specific molecules. Chondrocyte cell lines should be useful in studying the functions of these cells from normal and abnormal tissue and for pharmacological studies in vitro.

Diphtheria antitoxin levels in the 14-30-year age group in Italy.

In order to verify diphtheria immunity a seroepidemiological survey was performed in 1996-1997. Serum samples were obtained from 501 subjects 14 years old, recruited at 8 schools in Rome, and from 490 subjects 20-30 years old recruited from 15 Italian regions. Serum diphtheria antitoxin was titrated using the Vero cell assay. The minimum protection level of antitoxin was set at 0.01 IU ml-1. The results show that the younger population have a good immunity to diphtheria while a large proportion of young adults is devoid of protective levels of diphtheria antitoxin. Out of the 501 subjects 14 years old, 495 (98.8%) had a diphtheria antitoxin titre > or = 0.01 IU ml(-1). Only 6 (1.2%) teenagers were susceptible. Out of the 490 subjects 20-30 years old, 109 (22.2%) were susceptible, 381 (77.8%) had a diphtheria antitoxin titre > or = 0.01 IU ml(-1). The data stress for booster immunization at the end of junior high school.