Francesca Martucci

IZS Istituto Zooprofilattico Sperimentale, Teramo, Abruzzo, Italy

Are you Francesca Martucci?

Claim your profile

Publications (16)38.29 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Introduction - Dioxins are a family of organic micro-pollutants, highly toxic to man, able to persist long time in the environment, accumulate in the fat of humans and animals, and thus enter the food chain. The cell bioassay DR-CALUX (Dioxin Responsive Chemically Activated Luciferase Gene Expression), based on the mechanism of uptake of cellular receptor AhR for dioxins and dioxin-like compounds, is a method that has spread in recent years in many countries, as test of choice for screening of food, because of the relatively short run times and reduced costs compared to the confirmatory method, the high resolution gas chromatography and mass spectrometry (HRGC/HRMS). Aim - The purpose of this paper is to assess the applicability of the BDS-DR-CALUX® method as a screening tool for the monitoring of dioxins in milk produced in the Piedmont region, verifying the reliability compared to the data obtained through the official confirmatory method. Materials and method - Thirteen raw milk samples, previously examined by HRGC/HRMS, coming from a contaminated site near a steelworks in the area of Susa Valley, and 30 milk samples coming from the market distribution, were subjected to extraction of fats and subsequent purification into silica columns. For the dioxins determination, genetically modified hepatoma cells were cultured and incubated in 96 well plates and then left in contact for 24 hours with the extracts of the samples. Luminescence was measured with a chemiluminometer, exploiting the interaction mechanism between luciferine and luciferase. Results and discussion - The results obtained showed good infraclass correlation between the traditional method and the BDS-DR-CALUX®, in milk, a common food, especially in childhood nutrition, and used for further transformations. BDS -DR-CALUX® and chemical method can be considered as two valid systems to be used in series and are able to provide comparable results with regard to the relative quantification of dioxins: DR-CALUX gives a biological response of total mixture and allow to discriminate in advance through samples, which don't require an additional method of confirmation, and suspect samples, which will need to be confirmed by HRGC/HRMS, while HRGC/HRMS provides the concentration of specific chemical compounds in the mixture. Conclusions - BDS-DR-CALUX®, therefore, is confirmed as a valid screening method for milk, quick, fast, relatively cheap and able to allow a much broad monitoring of any areas potentially at risk for the population. Therefore the method allows for environmental monitoring and epidemiology of a high number of samples and test the contamination levels of dioxin in the food chain.
    No preview · Article · Jun 2013 · Large Animal Review
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Prion protein (PrP) is encoded by the PRNP gene, which is highly polymorphic in goats, with polymorphisms encoding amino acid substitutions at the protein level. In the current study, the reactivity of monoclonal antibody (mAb) F99/97.6.1 in binding PrP from goats polymorphic at PRNP codon 222 was investigated. Nervous tissue from 30 scrapie-negative goats with 3 different genotypes (222Q/Q, 222Q/K, and 222K/K) was analyzed by Western blot using mAbs P4 and F99/97.6.1. Although PrP was detected in all 30 samples by mAb P4, detection of PrP by mAb F99/97.6.1 was limited to 222Q/Q (12/12). No PrP was detected by mAb F99/97.6.1 in the 222K/K samples (n = 6), and the signal intensity of mAb F99/97.6.1 for PrP was lower for the 222Q/K samples (12/12 samples). To further investigate these results, additional Western blot analyses were performed, and the PrP signals detected by mAbs F99/97.6.1 and SAF84 were then quantified. The mean F99/SAF84 ratio (± standard deviation) calculated for the 222Q/Q group was 0.73 ± 1.26, and the mean for the 222Q/K group was 0.27 ± 1.31. Statistical analysis of these values evidenced statistically significant differences between the 222Q/Q and 222Q/K samples. The results of the study thus revealed an inhibition by lysine at position 222 on the binding of mAb F99/97.6.1 to goat PrP. This has implications for the use of mAb F99/97.6.1 for diagnostic purposes. Because the 222K allele could be a target for genetic selection in goats, the differential reactivity of mAb F99/97.6.1 could be exploited with a genotyping test setup.
    Full-text · Article · Sep 2012 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: ABSTRACT: Susceptibility of sheep to scrapie, a transmissible spongiform encephalopathy of small ruminants, is strongly influenced by polymorphisms of the prion protein gene (PRNP). Breeding programs have been implemented to increase scrapie resistance in sheep populations; though desirable, a similar approach has not yet been applied in goats. European studies have now suggested that several polymorphisms can modulate scrapie susceptibility in goats: in particular, PRNP variant K222 has been associated with resistance in case-control studies in Italy, France and Greece. In this study we investigated the resistance conferred by this variant using a natural Italian goat scrapie isolate to intracerebrally challenge five goats carrying genotype Q/Q 222 (wild type) and five goats carrying genotype Q/K 222. By the end of the study, all five Q/Q 222 goats had died of scrapie after a mean incubation period of 19 months; one of the five Q/K 222 goats died after 24 months, while the other four were alive and apparently healthy up to the end of the study at 4.5 years post-challenge. All five of these animals were found to be scrapie negative. Statistical analysis showed that the probability of survival of the Q/K 222 goats versus the Q/Q 222 goats was significantly higher (p = 0.002). Our study shows that PRNP gene mutation K222 is strongly associated with resistance to classical scrapie also in experimental conditions, making it a potentially positive target for selection in the frame of breeding programs for resistance to classical scrapie in goats.
    Full-text · Article · Feb 2012 · Veterinary Research
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: ABSTRACT: Three distinct forms of bovine spongiform encephalopathy (BSE), defined as classical (C-), low (L-) or high (H-) type, have been detected through ongoing active and passive surveillance systems for the disease.The aim of the present study was to compare the ability of two sets of immunohistochemical (IHC) and Western blot (WB) BSE confirmatory protocols to detect C- and atypical (L- and H-type) BSE forms.Obex samples from cases of United States and Italian C-type BSE, a U.S. H-type and an Italian L-type BSE case were tested in parallel using the two IHC sets and WB methods. The two IHC techniques proved equivalent in identifying and differentiating between C-type, L-type and H-type BSE. The IHC protocols appeared consistent in the identification of PrPSc distribution and deposition patterns in relation to the BSE type examined. Both IHC methods evidenced three distinct PrPSc phenotypes for each type of BSE: prevailing granular and linear tracts pattern in the C-type; intraglial and intraneuronal deposits in the H-type; plaques in the L-type.Also, the two techniques gave comparable results for PrPSc staining intensity on the C- and L-type BSE samples, whereas a higher amount of intraglial and intraneuronal PrPSc deposition on the H-type BSE case was revealed by the method based on a stronger demasking step.Both WB methods were consistent in identifying classical and atypical BSE forms and in differentiating the specific PrPSc molecular weight and glycoform ratios of each form. The study showed that the IHC and WB BSE confirmatory methods were equally able to recognize C-, L- and H-type BSE forms and to discriminate between their different immunohistochemical and molecular phenotypes. Of note is that for the first time one of the two sets of BSE confirmatory protocols proved effective in identifying the L-type BSE form. This finding helps to validate the suitability of the BSE confirmatory tests for BSE surveillance currently in place.
    Full-text · Article · Sep 2011 · BMC Research Notes
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Nor98 is an atypical scrapie strain characterized by a molecular pattern and brain distribution of the pathological prion protein (PrP(Sc)) different from classical scrapie. In Italy, 69 atypical cases have been identified so far and all were characterized as Nor98 strain. In this paper we report an unusual case in a sheep which showed immunohistochemical and molecular features of PrP(Sc) different from the other atypical cases. The sheep was from an outbreak where the index and the other four cases were affected by classical scrapie. Histopathological, immunohistochemical and Western blot analyses on the brain of the unusual case revealed the simultaneous presence of pathological features characteristic of Nor98 and classical scrapie. Interestingly, the prevalent disease phenotype in the brainstem was classical scrapie-like, while in the cerebral cortex and cerebellum the Nor98 phenotype was dominant. The sub-mandibular lymph node was positive and showed a PrP(Sc) molecular pattern referable to classical scrapie. The PrP genotype was AL(141)RQ/AF(141)RQ. Taken together, the occurrence of classical scrapie in the outbreak, the PrP genotype, the involvement of different cellular targets in the brain and the pathological and molecular PrP(Sc) features observed suggest that this unusual case may result from the co-existence of Nor98 and classical scrapie.
    Full-text · Article · Jun 2010 · Research in Veterinary Science
  • [Show abstract] [Hide abstract]
    ABSTRACT: The olfactory system (OS) involvement in transmissible spongiform encephalopathies has lately been acknowledged in experimental studies: Prion spread to the nasal mucosa is known to occur in hamsters after intracerebral challenge, and olfactory neurons have been assessed as a route for prion neuroinvasion too. This study investigated whether the OS is involved in naturally occurring prion diseases. Samples of nasal mucosa taken at the level of medial nasal concha, ventral nasal concha and nasal septum from 24 natural scrapie affected sheep were examined by immunohistochemistry (IHC), immunofluorescence (IF), PET blot and Western Blot (WB) for scrapie prion protein (PrPSc). OS related brain areas of the selected sheep (olfactory bulb, olfactory tract, frontal cortex, pyriform lobe and hippocampus) were analyzed too. Prion spread was assessed both in peripheral and central OS of the examined sheep. Twenty one samples of olfactory mucosa were positive by WB; IHC confirmed WB positive results in 14 cases. PrPSc was mainly localized in the medial nasal concha at the level of the olfactory nerve perineurium, and it was also disclosed in the nasal associated lymphoid tissue. It was never detected in olfactory neurons and fibers. In the brain PrPSc staining intensity was higher in olfactory bulb and frontal cortex, where it appeared as submeningeal, subependymal and perivascular patterns. The finding of PrPSc both in the brain areas directly in contact to the cerebrospinal fluid (CSF) and in the olfactory nerve perineurium bounding the subdural space extension that surrounds nerve rootlets might be consistent with the recently discovered PrPSc presence in CSF.
    No preview · Article · Feb 2010 · Large Animal Review
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: To evaluate further the reactivity of prion-specific monoclonal antibodies containing the 89-112 or 136-158 prion protein (PrP) polypeptides, immunoprecipitations were performed on brain extracts from Italian bovines, sheep and goats with transmissible spongiform encephalopathies. No binding of IgG 89-112 or IgG 136-158 to PrP in normal brain extracts was detected. Conversely, both reagents immunoprecipitated PrP from bovine and bovine amyloidotic spongiform encephalopathies, and from typical and atypical scrapie brain extracts. The immunoprecipitated PrP bands mirrored the Western blot (WB) profile of the different prion strains, indicating universal affinity of two independent PrP regions for disease-associated PrP conformers regardless of species source and strain properties. Immunoprecipitation with motif-grafted antibodies increased the sensitivity of conventional detection methods based on centrifugation followed by WB, which was confirmed by assay of diluted samples using both methods. These reagents or derivative molecules may thus find broad applications in prion detection and research.
    Full-text · Article · Mar 2009 · Journal of General Virology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The olfactory system (OS) is involved in many infectious and neurodegenerative diseases, both human and animal, and it has recently been investigated in regard to transmissible spongiform encephalopathies. Previous assessments of nasal mucosa infection by prions following intracerebral challenge suggested a potential centrifugal spread along the olfactory nerve fibers of the pathological prion protein (PrPSc). Whether the nasal cavity may be a route for centripetal prion infection to the brain has also been experimentally studied. With the present study, we wanted to determine whether prion deposition in the OS occurs also under field conditions and what type of anatomical localization PrPSc might display there. We report here on detection by different techniques of PrPSc in the nasal mucosa and in the OS-related brain areas of sheep affected by natural scrapie. PrPSc was detected in the perineurium of the olfactory nerve bundles in the medial nasal concha and in nasal-associated lymphoid tissue. Olfactory receptor neurons did not show PrPSc immunostaining. PrPSc deposition was found in the brain areas of olfactory fiber projection, chiefly in the olfactory bulb and the olfactory cortex. The prevalent PrPSc deposition patterns were subependymal, perivascular, and submeningeal. This finding, together with the discovery of an intense PrPSc immunostaining in the meningeal layer of the olfactory nerve perineurium, at the border with the subdural space extension surrounding the nerve rootlets, strongly suggests a probable role of cerebrospinal fluid in conveying prion infectivity to the nasal submucosa.
    Full-text · Article · Feb 2009 · Journal of Virology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Bovine spongiform encephalopathy (BSE), the prion disease in cattle, was widely believed to be caused by only one strain, BSE-C. BSE-C causes the fatal prion disease named new variant Creutzfeldt-Jacob disease in humans. Two atypical BSE strains, bovine amyloidotic spongiform encephalopathy (BASE, also named BSE-L) and BSE-H, have been discovered in several countries since 2004; their transmissibility and phenotypes in humans are unknown. We investigated the infectivity and human phenotype of BASE strains by inoculating transgenic (Tg) mice expressing the human prion protein with brain homogenates from two BASE strain-infected cattle. Sixty percent of the inoculated Tg mice became infected after 20 to 22 months of incubation, a transmission rate higher than those reported for BSE-C. A quarter of BASE strain-infected Tg mice, but none of the Tg mice infected with prions causing a sporadic human prion disease, showed the presence of pathogenic prion protein isoforms in the spleen, indicating that the BASE prion is intrinsically lymphotropic. The pathological prion protein isoforms in BASE strain-infected humanized Tg mouse brains are different from those from the original cattle BASE or sporadic human prion disease. Minimal brain spongiosis and long incubation times are observed for the BASE strain-infected Tg mice. These results suggest that in humans, the BASE strain is a more virulent BSE strain and likely lymphotropic.
    Full-text · Article · May 2008 · Journal of Virology
  • Source
    P L Acutis · F Martucci · M Mazza · S Nodari · C Maurella · G Ru · C Casalone · M Caramelli

    Full-text · Article · Dec 2006 · The Veterinary record
  • [Show abstract] [Hide abstract]
    ABSTRACT: Susceptibility to scrapie in sheep depends on the host PrP genotype. No data about the linkage of the rare ARK allele to differential scrapie susceptibility are currently available. Several tissues isolated from sheep from an Italian scrapie outbreak and carrying the ARK allele were examined for the presence of the pathological prion protein. A weak positivity was detected only by Western blot in the brainstem of one ARK/ARH sheep. This result shows that the ARK allele does not confer full resistance against scrapie and that the allele needs to be studied further before it can be considered for breeding purposes.
    No preview · Article · Oct 2006 · Archives of Virology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Without Abstract
    Full-text · Article · Jul 2006 · Veterinary Research Communications
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Tongue involvement by prion spreading was shown to be a common outcome after oral or intracranial experimental challenge with scrapie and transmissible mink encephalopathy sources in rodent models. It is also known that bovine spongiform encephalopathy, which is pathogenic for humans, is experimentally transmissible to sheep and can lead to a disease indistinguishable from scrapie. A recent European Food Safety Authority opinion recommended research into PrPsc accumulation in the tongues of ruminants. We report on the detection of PrPsc in the tongues of seven scrapie-infected sheep by immunohistochemistry and Western blotting.
    Full-text · Article · Jun 2005 · Journal of Virology
  • [Show abstract] [Hide abstract]
    ABSTRACT: In accordance with EU Regulation 999/2001, rapid tests already adopted for bovine spongiform encephalopathy (BSE; Prionics Check Western, Platelia-BSE and Enfer TSE) are to be applied in all European countries to a sub-population of over 18-month-old slaughtered or dead sheep and goats to improve Scrapie surveillance and to determine the possible presence of BSE in sheep; however, the three tests have thus far been evaluated only for BSE and no official data are available about their performances on Scrapie. We evaluated the accuracy of these methods for TSE diagnosis in Italian sheep and goats, using a pre-homogenisation protocol on brain-stem samples to obtain comparable data from the three tests. Our results show that the tests can be considered reliable tools for active surveillance in the small ruminants population.
    No preview · Article · Jul 2004 · Acta Neuropathologica
  • Source

    Full-text · Article ·
  • Source
    F. Martucci · M. Mazza · S. Nodari · C. Maurella · G. Ru · C. Casalone

    Full-text · Article ·