Loredana Ingrosso

Sulphate polyanions prolong the incubation period of scrapie-infected hamsters

The effect of the organic sulphated polyanions, pentosan sulphate (SP54), dextran sulphate 500 (DS500) and suramin, have been tested on golden Syrian hamsters infected with the 263K strain of scrapie by the intraperitoneal (i.p.) or the intracerebral route. SP54 had the greatest effect in prolonging the incubation period of the disease when administered within 2 h of the i.p. inoculum. The same amount of SP54 given 24 h after scrapie inoculation had a potent effect in some animals and no effect in others. This result suggests that SP54 inhibits the uptake of the scrapie agent into the nerve endings and/or carrier cells at the site of the inoculum, i.e. the peritoneum, and that this event occurs in about 24 h. DS500 had a similar although less potent effect (22.4 days delay during the incubation period) than SP54 (54.4 days) when administered within 2 h of scrapie injection by the i.p. route, and suramin had only a minimal effect (10 days). This study suggests that treatment of scrapie and related spongiform encephalopathies of animals and man is possible only before the agent has reached the clinical target areas of the brain.

Transmission of the 263K scrapie strain by the dental route.

Apart from a few cases of iatrogenic and familial human transmissible spongiform encephalopathies (TSEs) or prion diseases, the cause of Creutzfeldt-Jakob disease (CJD) remains unknown. In this paper we investigated the possibility that dental procedures may represent a potential route of infection. This was assessed by using the experimental model of scrapie in hamster. In the first part of this study we found that after intraperitoneal inoculation, oral tissues commonly involved in dental procedures (gingival and pulp tissues) bore a substantial level of infectivity. We also found high scrapie infectivity in the trigeminal ganglia, suggesting that the scrapie agent had reached the oral tissues through the sensitive terminal endings of the trigeminal nerves. In the second part of the study we inoculated a group of hamsters in the tooth pulp and showed that all of them developed scrapie disease. In these animals, we detected both infectivity and the pathological prion protein (PrPsc) in the trigeminal ganglion homolateral to the site of injection but not in the controlateral one. This finding suggests that the scrapie agent, and likely other TSE agents as well, spreads from the buccal tissues to the central nervous system through trigeminal nerves. Although these findings may not apply to humans affected by TSEs, they do raise concerns about the possible risk of transmitting these disorders through dental procedures. Particular consideration should be taken in regard to new variant CJD patients because they may harbour more infectivity in peripheral tissues than sporadic CJD patients.

Chronic wasting disease and atypical forms of bovine spongiform encephalopathy and scrapie are not transmissible to mice expressing wild-type levels of human prion protein

The association between bovine spongiform encephalopathy (BSE) and variant Creutzfeldt-Jakob disease (vCJD) has demonstrated that cattle transmissible spongiform encephalopathies (TSEs) can pose a risk to human health and raises the possibility that other ruminant TSEs may be transmissible to humans. In recent years, several novel TSEs in sheep, cattle and deer have been described and the risk posed to humans by these agents is currently unknown. In this study, we inoculated two forms of atypical BSE (BASE and H-type BSE), a chronic wasting disease (CWD) isolate and seven isolates of atypical scrapie into gene-targeted transgenic (Tg) mice expressing the human prion protein (PrP). Upon challenge with these ruminant TSEs, gene-targeted Tg mice expressing human PrP did not show any signs of disease pathology. These data strongly suggest the presence of a substantial transmission barrier between these recently identified ruminant TSEs and humans.

Molecular diagnostics of transmissible spongiform encephalopathies

Clinical criteria for the diagnosis of sporadic, iatrogenic and variant Creutzfeldt-Jakob diseases are now available and show an excellent sensitivity and specificity ( approximately 98%). Post-mortem diagnosis, based upon the identification in the brain of the pathological conformer of the prion protein (PrP(Sc)), is also very accurate, and several diagnostic kits are now available that facilitate the immunochemical measurement of PrP(Sc). Several new molecular diagnostic techniques aimed at increasing the sensitivity and specificity of PrP(Sc) detection, and at identifying markers of disease that are other than PrP(Sc), are the subject of ongoing studies. The aim of these studies is to develop preclinical screening tests for the identification of infected, but still healthy, individuals. These tests are also badly needed to check the safety of blood or blood-derived products, and to ensure meat safety in European countries.

Risk factors for tuberculosis in foreign-born people (FBP) in Italy: a systematic review and meta-analysis.

In Italy, TB notifications in foreign-born people (FBP) are steadily increasing. To investigate this issue we did a meta-analysis on risk factors for FBP people. A systematic search was performed in PubMed and EMBASE from Jan-1980 to Jan-2013. We analysed HIV status, previous TB-treatment, intravenous drug use and alcohol abuse, and multidrug resistant TB. Odd ratio was used as a measure of effect. One and two-stages approaches were used. In the main analysis we used a 2-stages approach to include studies with only aggregate estimates. Among 1996 references, 18 fulfilled inclusion criteria. In TB-affected FBP people positive HIV-status was about 3 times higher than among Italians, after 1996 when combined antiretroviral therapy for HIV was introduced (OR: 2.91; 95%CI: 1.37; 6.17). No association was found between FBP and intravenous drug users in adults; after 1-stage meta-analysis foreign born people from highly endemic countries had a 4 times higher risk to be multidrug resistant TB than Italian people. Finally, TB-affected FBP were less likely than Italians to be alcoholics (OR: 0.10 95%CI: 0.01; 0.84) or of having received previous TB-treatment (OR: 0.55; 95%CI: 0.43; 0.71). An association of multidrug resistant TB with immigrant status as well as an association of Tuberculosis with HIV-positive status in foreign-born people are major findings of this analysis. Drugs and alcohol abuse do not appear to be risk factors for TB in FBP, however they cannot be discharged since may depend on cultural traditions and their role may change in the future along with the migratory waves. An effective control of TB risk factors among migrants is crucial to obtain the goal of TB eradication.

Mycobacterium smegmatis Expressing a Chimeric Protein MPT64-Proteolipid Protein (PLP) 139–151 Reorganizes the PLP-Specific T Cell Repertoire Favoring a CD8-Mediated Response and Induces a Relapsing Experimental Autoimmune Encephalomyelitis

We infected SJL mice with a recombinant Mycobacterium smegmatis expressing a chimeric protein containing the self-epitope of proteolipid protein 139–151 (p139) fused to MPT64, a secreted protein of Mycobacterium tuberculosis (rMSp139). Infected mice developed a relapsing experimental autoimmune encephalomyelitis (EAE), showing a prevailing demyelination of the CNS, and disease severity was significantly lower in comparison with the one that follows immunization with p139. rMSp139 was not detected in lymph node or spleen in the course of clinical disease development or in the CNS during relapse. Infection with rMSp139 modified the p139-specific T cell repertoire, recruiting the spontaneous p139-specific repertoire and activating CD4+ T cells carrying the BV4 semiprivate rearrangement. T cells carrying the public BV10 rearrangement that are consistently found in the CNS during flares of disease were not activated by infection with rMSp139 because lymph node APCs infected with rMSp139 selectively fail to present the epitope for which BV10 cells are specific. Simultaneously, rMSp139 expanded p139-specific CD8+ cells more efficiently than immunization with peptide in adjuvant. SJL mice vaccinated against the CDR3 sequence of the BV10 public rearrangement reduced usage of the BV10 cells and displayed reduced symptoms during bouts of EAE. Thus, transient peripheral infection with a CNS-cross–reactive nonpathogenic Mycobacterium induces a relapsing EAE that continues long after clearance of the infectious agent. The composition of the self-reactive repertoire activated determines severity and histology of the resulting disease.

Prion Strain Characterization of a Novel Subtype of Creutzfeldt-Jakob Disease

ABSTRACT In 2007, we reported a patient with an atypical form of Creutzfeldt-Jakob disease (CJD) heterozygous for methionine-valine (MV) at codon 129 who showed a novel pathological prion protein (PrPTSE) conformation with an atypical glycoform (AG) profile and intraneuronal PrP deposition. In the present study, we further characterize the conformational properties of this pathological prion protein (PrPTSE MVAG), showing that PrPTSE MVAG is composed of multiple conformers with biochemical properties distinct from those of PrPTSE type 1 and type 2 of MV sporadic CJD (sCJD). Experimental transmission of CJD-MVAG to bank voles and gene-targeted transgenic mice carrying the human prion protein gene (TgHu mice) showed unique transmission rates, survival times, neuropathological changes, PrPTSE deposition patterns, and PrPTSE glycotypes that are distinct from those of sCJD-MV1 and sCJD-MV2. These biochemical and experimental data suggest the presence of a novel prion strain in CJD-MVAG. IMPORTANCE Sporadic Creutzfeldt-Jakob disease is caused by the misfolding of the cellular prion protein, which assumes two different major conformations (type 1 and type 2) and, together with the methionine/valine polymorphic codon 129 of the prion protein gene, contribute to the occurrence of distinct clinical-pathological phenotypes. Inoculation in laboratory rodents of brain tissues from the six possible combinations of pathological prion protein types with codon 129 genotypes results in the identification of 3 or 4 strains of prions. We report on the identification of a novel strain of Creutzfeldt-Jakob disease isolated from a patient who carried an abnormally glycosylated pathological prion protein. This novel strain has unique biochemical characteristics, does not transmit to humanized transgenic mice, and shows exclusive transmission properties in bank voles. The identification of a novel human prion strain improves our understanding of the pathogenesis of the disease and of possible mechanisms of prion transmission.

Effect of Amphotericin B on Different Experimental Strains of Spongiform Encephalopathy Agents

Scrapie is a transmissible disease of the central nervous system (CNS) naturally occurring in sheep and goats, characterized by the formation of a modified, partly proteinase-resistant protein of the host, which tends to aggregate as amyloid fibrils and accumulate in the brain of infected individuals. We have previously reported that treatment with the polyene antibiotic amphotericin B (AmB) prolongs the incubation period of hamsters infected intracerebrally or intraperitoneally with strain 263K of scrapie. In this chapter, we show that AmB also prolongs the incubation periods of Armenian hamsters experimentally infected with the 263K strain of scrapie but not in Chinese hamsters. We also report the beneficial effect of AmB in mice injected with the KFu strain of CJD.

A desk review on institutional and non-institutional organizations active in the field of migrant's health in the WHO European Region

BACKGROUND Migrants have problematic access to health-care; non-institutional organizations (NGOs), as well as institutional bodies may play a role in facilitating their access to mainstream health care. AIM Our research reviews actions that address the need of migrants in terms of health care in order to understand how, where, and who participates in this effort. METHOD Data were from desk or web research, declaration from organisations and their websites, information from WHO Country Offices. RESULTS 154 NGOs were identified in the WHO European Region. 58% were direct health care providers while the remaining provided either mediation services or were part of a network organization. 173 national institutes (GOVs) were found; less than the 20% were directly or indirectly involved in health care, whereas the majority were involved in research, policy development, international relations and human rights. CONCLUSION AND RECOMMENDATION Some gaps, a certain fragmentation and lack of coordination were identified. WHO can play an overarching role in the exchange of expertise and harmonisation of the efforts in this field.

The CNCCS, a benchmark Italian consortium for bioeconomy and an opportunity for the Istituto Superiore di Sanità

BioEconomy is the word for a recently arisen paradigm that constitutes the inspirational principle of CNCCS (Collezione Nazionale Composti Chimici e Centro Screening – National Collection of Chemical Compounds and Screening Centre), a newly publicprivate partnership in Italy. BioEconomy postulates that science can be safely coupled with economy and that from such “unritual marriage” would derive a benefit for the society triggering welfare through innovation. The CNCCS is composed by the National Research Council (CNR), the temple of Italian research, the Italian National Institute of Health (ISS), the leading research institution of the National Health System, and the IRBM Science Park, a private company with the most advanced technology for exploiting organic compounds. The CNCCS is funded by the Italian Ministry of Education, University and Research (MIUR) through the CNR to encourage a never-seen-before synergy between public research bodies and private institutions. The scientific power of CNR and ISS merging with IRBM’s resources for the identification and collection of chemicals will make the CNCCS the leading reference European centre for “orphan” compounds. Why such an effort? Why the ISS should mingle in a public private partnership? The role of basic research is under scrutiny and poorly tolerated when a depressed economy and stakeholders (patients, families, society in general) put great pressures to direct the objectives of science toward goals that must result in benefits for society. The ISS has a special responsibility because its role is to address the needs of society by taking up the challenges of science beyond the existing limits. The ISS foresees that the CNCCS consortium would better serve the scope of science and society, a goal that is unattainable by science or industry alone. Private-public enterprises are quickly rising and institutional research bodies are becoming partners of charities, large pharmaceutical companies, and small biotech firms for looking at novel and affordable drugs for the cure of neglected diseases. Examples of these enterprises are the Malaria Vaccine Initiatives, the Medicines for Malaria Ventures, the Global alliance for TB drug development, and the Drug for Neglected Diseases Initiatives. What would be the contribution of the CNCCS consortium for the society and the general welfare in terms of health and economy? The quality of science in Italy is higher than lay people think despite poor public and private investments. In Italy, particularly in the ISS, there is a long-standing tradition in the field of organic chemistry with tens of thousands molecules continuously synthesized in public and private research laboratories. Unfortunately, most of discovered molecules “sit on the shelf” of academic and scientific laboratories and are poorly or not tested for potential therapeutic applications, limiting their potential value. The joined effort of the CNCCS partners would guarantee an extended lifespan of the molecules entrusted in the repository and would serve as a national high throughput hub for the identification of novel lead compounds for targets of interest. In the CNCCS, the efficacy of collected molecules is investigated for scopes that are beyond their original intended use. This is obviously done with the agreement of the original inventor who retains the patent for the first intended purpose and gives the consortium the right of exploring any possible further use of the molecule. Entrusting the results of his/her research into a repository is certainly an act that requires a fresh attitude toward science, something that could be defined as a step away from the bench and a plunge into reality. The CNCCS commissioned an ad hoc study to Fabio Pammolli, an economist and director of the IMT institute for advanced studies in Lucca for investigating how life sciences impact on the territory by enhancing the economical growth of society and producing wealth. The study showed that despite a high rate of scientific publication (Italy ranks 4th in the world), our country lacks a parallel rate of production in industrial and technological research. at regional level, however, the study showed a positive effect of “translated science” since the number of publications and patents, which are indices of innovation outcomes, correlates with wealth and productivity (unpublished data). Thus, this study confirms the pressing need of knowledge integration and urges scientists and policy makers to bridge the gap between academia and industry. Scientists and pharmaceutical companies should break old schemes and seeking new ad more productive alliances. We are at a deadlock with scientists publishing thousands of “chemical scaffolds” that are far from being a “drug” and pharmaceutical companies reluctant to invest money in research of new leading drugs, because the pay back for investments is becoming more and more uncertain. Editorial