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SARS-CoV-2 serology in patients on biological therapy or apremilast for psoriasis: a study of 93 patients in the Italian red zone Finally, we used the serological data to determine the usefulness of an over-the-phone questionnaire for SARS-CoV-2 positivity performed by Brazzelli I et al i .3 on the same cohort of psoriatic patients. None of the 12 positive patients had a severe infection or required hospitalisation due to SARS-CoV-2 infection. [Extracted from the article]

Background:Breast cancer clinical outcome is affected by tumor molecular features, and the identification of subtype-specific prognostic biomarkers is relevant for breast cancer translational research. Gene expression signatures proved to be able to complement prognostic information provided by classical clinico-pathological features. Recently, microRNAs (miRNAs) have been causally linked to tumorigenesis and cancer progression and have been associated with patient outcome, also in breast cancer.Methods:MicroRNAs associated with the development of distant metastasis were identified in a cohort of 92 ESR1+/ERBB2− lymph node-negative breast cancers from patients not receiving adjuvant treatment. Results were confirmed and further investigated in a total of 1246 miRNA and gene expression profiles of the Molecular Taxonomy of Breast Cancer International Consortium data set. Moderated t-test, univariable and multivariable Cox regression models were used for statistical analyses.Results:miR-30e* was identified as independent protective prognostic factor in lymph node-negative untreated patients with ESR1+/ERBB2− tumours and retained a significant association with a good prognosis in treated patients with the same tumor subtype as well as in the ERBB2+ subtype, but not in ESR1−/ERBB2− tumours.Conclusions:We highlighted a relevant and subtype-specific role in breast cancer for miR-30e* and demonstrated that adding miRNA markers to gene signatures and clinico-pathological features can help for a better prognostication. [ABSTRACT FROM AUTHOR]

Abstract Background Gene expression profiling is moving from the research setting to the practical clinical use. Gene signatures able to correctly identify high risk breast cancer patients as well as to predict response to treatment are currently under intense investigation. While technical issues dealing with RNA preparation, choice of array platforms, statistical analytical tools are taken into account, the tissue collection process is seldom considered. The time elapsed between surgical tissue removal and freezing of samples for biological characterizations is rarely well defined and/or recorded even for recently stored samples, despite the publications of standard operating procedures for biological sample collection for tissue banks. Methods Breast cancer samples from 11 patients were collected immediately after surgical removal and subdivided into aliquots. One was immediately frozen and the others were maintained at room temperature for respectively 2, 6 and 24 hrs. RNA was extracted and gene expression profile was determined using cDNA arrays. Phosphoprotein profiles were studied in parallel. Results Delayed freezing affected the RNA quality only in 3 samples, which were not subjected to gene profiling. In the 8 breast cancer cases with apparently intact RNA also in sample aliquots frozen at delayed times, 461 genes were modulated simply as a function of freezing timing. Some of these genes were included in gene signatures biologically and clinically relevant for breast cancer. Delayed freezing also affected detection of phosphoproteins, whose pattern may be crucial for clinical decision on target-directed drugs. Conclusion Time elapsed between surgery and freezing of samples appears to have a strong impact and should be considered as a mandatory variable to control for clinical implications of inadequate tissue handling.

Abstract Background Dogs are associated with more than 60 zoonotic diseases among which, parasitosis and, in particular, helminthosis, can pose serious public-health concerns worldwide. Many canine gastrointestinal parasites eliminate their dispersion elements (eggs, larvae, oocysts) by the faecal route. The quantity of canine faeces deposited on public and private property in cities worldwide is both a perennial nuisance and an important health issue. Public sites such as playgrounds, parks, gardens, public squares and sandpits may be an important source of human infection. The aim of this study was to evaluate the extent of both canine faecal contamination in the city of Naples (southern Italy), and presence of canine parasitic elements, with particular regard to those which are potential agents of zoonosis. A regular grid of sub-areas (1 km × 700 m) was overlaid on the city map using a Geographical Information System (GIS). In each sub-area the straightest 1 km transect was drawn and digitalized on-screen in the GIS. Between February and May 2005 canine faeces were counted along the 1 km transects in 143 sub-areas, and 415 canine faecal samples were collected and submitted to coprological examinations. Negative binomial regression models and Gaussian random effects models were used to analyze the association between faeces count and human population density taking into account for extraPoisson variability. Logistic regression model was used to evaluate the association between positivity to parasitic elements and number of canine faeces. Results Out of the 143 studied sub-areas, 141 (98.6%) contained canine faeces. There was a strong spatial gradient with 48% of the total variability accounted by between neighbourhood variability; a positive association between the number of faeces and the human population density was found. Seventy (over 415, 16.9%) canine faecal samples were positive for parasitic elements. There was no association between positivity to parasitic elements and the number of canine faeces. Eggs of Toxocara canis, Toxascaris leonina, Ancylostoma caninum and Trichuris vulpis were found, as well as oocysts of Isospora canis. Conclusion In conclusion, the results of the present study, conducted using GIS both for planning and sampling and for evaluation and presentation of findings, showed the presence of canine faecal contamination in the city of Naples, and the presence of canine parasitic elements, some of which are potential agents of zoonosis.

Abstract: The FLOTAC techniques involve the spinning of faecal samples onto the surface of counting chambers to permit enumeration of parasitic elements (eggs, larvae, oocysts and cysts) to an accuracy of one parasitic element per gram of faeces. In the present study it is demonstrated that FLOTAC provides a rapid and very sensitive method for counting of lungworm larvae of sheep. The optimum flotation solution for lungworm larvae is zinc sulphate and mercury II iodide (s.g. 1.45) although zinc sulphate (s.g. 1.20 or 1.35) on its own also gave good results. Samples preserved in 5% formalin gave the highest counts but fresh, frozen and samples in 10% formalin also gave higher counts than McMaster and simple flotation. Larval counts of 307 field samples gave up to 1.27x more positives samples than use of Baermann funnels and up to 4.18x more larvae per sample. As FLOTAC is faster than Baermannisation of samples it offers a better method of counting larvae in ruminant faecal samples. [Copyright &y& Elsevier]

Abstract: A cross-sectional serological survey was conducted on cattle pasturing in an area of the southern Italian Apennines to evaluate the seroprevalence to Neospora caninum, and to investigate the climatic, environmental, farm management, and individual animal factors that influence the distribution of this protozoan. Blood samples were collected from 864 pastured cattle raised on 81 farms. Serum samples were tested for antibodies to N. caninum using an ELISA assay (HerdCheck®, IDEXX). A geographical information system (GIS) for the study area was constructed using the following remote sensing (RS) and landscape feature data: autumn–winter, spring and summer normalized difference vegetation index (NDVI), land cover, elevation, slope, aspect, mean rainfall and minimum, mean, and maximum temperature in spring, summer, autumn and winter. Data on each of these features were then extracted for “buffer zones” consisting of the area included in a circle of 3km diameter centered on the 81 geo-referenced centroids of the main cattle pastures. Climatic and environmental data obtained from RS and GIS and individual animal characteristics and farm management data obtained from a questionnaire were analyzed in relation to N. caninum seropositivity and antibody titres both by linear and logistic regression models. Out of the 81 farms sampled, 63 (77.8%) had at least one tested animal positive for N. caninum. Out of the 864 bovine sera samples, 266 (30.8%) were found to have antibodies to N. caninum. The results of the logistic regression model show that significantly high seroprevalence to N. caninum was found in heifers/steers and adults, in cattle raised on farms having a large number of dogs, and in cattle raised in buffer zones having high minimum temperatures in the spring, and a narrow extension of summer NDVI. In addition, positive linear correlations were found between N. caninum antibody titres and the number of dogs on farm, and the minimum temperature in spring. All the above determined risk factors for N. caninum seroprevalence indicate that horizontal infection resulting from the ingestion of oocysts shed by dogs is the most probable route of N. caninum infection in pastured cattle of the southern Italian Apennines. [Copyright &y& Elsevier]

The aims of this research were to evaluate mean echogenicity (ME) of the deep and superficial digital flexor tendons (DDFT and SDFT), the interosseous muscle (IM), and the accessory ligament of the deep digital flexor tendon (ALDDFT) of the metacarpal region in neonatal foals, and determine the effect of sex, side and body weight on this quantitative ultrasonographic evaluation. Thirteen orthopedically sound neonatal foals were examined. Four areas of study (1A, 1B, 2A, 2B) were identified. Transverse scans of the DDFT, SDFT, IM and ALDDFT were obtained, recorded, and analyzed. The most echogenic structures were the ALDDFT and DDFT, while the SDFT was significantly less echogenic than all other structures (P < 0.05). No influence of sex, forelimb, or body weight was observed. The echogenicity of the tenodesmic structures of foals partially overlapped that reported in the metacarpal region in adult horses, except for IM. [ABSTRACT FROM AUTHOR]

Abstract: The present study was aimed at carrying out a cross-sectional copromicroscopic survey of helminths and intestinal protozoa in immigrants in Naples (southern Italy). Between October 2008 and November 2009, a total of 514 immigrants were tested comparing the FLOTAC dual technique and the ethyl acetate concentration technique. Combined results of the two techniques served as a diagnostic ‘gold’ standard and revealed an overall prevalence of parasitic infections of 61.9% (318/514). The ethyl acetate concentration technique detected a low number of positive results (49.0%) and this was confirmed for each helminth/protozoa species detected. Among helminths, Trichuris trichiura (3.9%), hookworms (3.7%) and Ascaris lumbricoides (1.4%) were the most prevalent. Strongyloides stercoralis (0.4%), Enterobius vermicularis (0.4%), Schistosoma mansoni (1.0%), Hymenolepis nana (1.6%) and Taenia spp. (0.2%) were also found, as well as zoonotic helminths, as Trichostrongylus spp. (0.8%) and Dicrocoelium dendriticum (0.8%). As regard to pathogenic protozoa, Blastocystis hominis was the most commonly detected (52.7%), followed by Entamoeba histolytica/Entamoeba dispar/Entamoeba moshkovskii (11.9%) and Giardia duodenalis (4.5%). Several issues concerning diagnosis, epidemiology and public health impact of parasitic infections in immigrants are offered for discussion. In conclusion, the present paper pointed out the need of better diagnosis and cure of the immigrants population in order to improve access to health care of this neglected and marginalised population group, for its own protection and care. [Copyright &y& Elsevier]

Abstract: Gastrointestinal (GI) strongyle infections remain one of the main constraints to goat production worldwide. Although several indicators of GI strongyle infections have been proposed, faecal egg count (FEC) techniques are the most common approaches for the estimation of prevalence and intensity of these parasites. However, FEC may be subjected to a within-individual variation due to factors such as weather, season, random day-to-day variation and phase of the parasitic infection. A longitudinal study of GI strongyle FEC was conducted on 63 dairy goats to evaluate the effects of hour of sample collection on FEC and the relationship between FEC and worm burden. The goats were naturally infected with Teladorsagia circumcincta, Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum venulosum. Every 3 weeks for 14 months faeces were collected every 2h for 24h from three individually caged goats. The faeces were those passed by the goats during the 2h preceding each collection. For each goat a sample from this material was analyzed using the Flotac double technique. To explore relationships between FECs and adult parasite counts, on the day following sampling, the three goats were euthanized and the nematodes present in the abomasa and intestines were recovered, identified and counted. The results of the general linear model did not show any significant effect of the hour of faecal sample collection on GI strongyle FEC, whereas a significant effect of the month of faecal sample collection was found. The results of the Pearson correlation showed a significant (P =0.000) positive relationship between FEC and worm burden (r =0.6), in particular regarding H. contortus (r =0.9). In conclusion, the present study showed that the hour of sample collection does not influence the GI strongyle FEC and that there is a good relationship between FEC and total GI strongyle worm burden in goats. [Copyright &y& Elsevier]

Abstract: A survey of cystic echinococcosis (CE) in the water buffalo (Bubalus bubalis) of the Italian Mediterranean breed was carried out in Campania, a region of southern Italy. In addition, a molecular study was performed on 48 hydatid cysts coming from 48 water buffaloes in order to determine the Echinococcus granulosus strain(s) present in this host. Out of a total of 722 water buffaloes examined for CE, 76 (10.5%) were found infected. The average number of cysts per buffalo was 4.3 (minimum 1, maximum 45). Seventeen buffaloes had hydatid cysts only in the liver (with an average of 5cysts/liver), 34 only in the lungs (with an average of 1.8cysts/lungs), and 25 buffaloes had cysts both in the liver and in the lungs. Fertile cysts were found in 10 (13.2%) out of the 76 positive buffaloes. The sequencing of the mitochondrial cytochrome C oxidase subunit 1 (CO1) gene of the 48 hydatid cysts produced sequences of 419bp for each sample analysed. For 33 samples, alignment of the obtained sequences with those present in GenBank showed a total homology with the common domestic sheep strain G1; for 15 samples, sequences obtained showed 100% homology with buffalo strain G3. The findings of the present survey represent the first epidemiological and molecular comprehensive studies on CE in water buffalo from an endemic area for E. granulosus. [Copyright &y& Elsevier]

Abstract: Isolates of the rumen fluke Calicophoron daubneyi (Digenea: Paramphistomidae) from various hosts and three locations in southern Italy were characterized genetically. The second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA) plus flanking 5.8S and 28S sequence (ITS-2+) was amplified from individual rumen flukes by PCR. PCR-linked restriction fragment length polymorphism (PCR-RFLP) analysis was performed using four different restriction endonucleases, and PCR products were sequenced. The PCR analyses from all the C. daubneyi specimens produced identical fragments, and the PCR-RFLP analyses did not show, with respect to any of the four restriction endonucleases, any differences between the C. daubneyi specimens. The sequence analyses of the ITS-2+ from each of the C. daubneyi specimens showed them all to be 428bp, and composed of the entire ITS-2 sequence (282bp) plus the two partial flanking conserved sequences, 5.8S (99bp) and 28S (47bp). No intra-specific variation was observed in the nucleotide composition of the ITS-2+ (homology=100%). There was, however, an observable inter-specific variation between the ITS-2+ of C. daubneyi and the ITS-2+ of both Calicophoron calicophorum (homology=97.2 %) and Calicophoron microbothrioides (homology=97.4 %), both previously deposited in the GenBank™. The finding of the present study shows that, as has already demonstrated for other parasitic helminths, ITS-2 can serve as an effective genetic marker for the molecular identification of paramphistomes, and as a useful tool for developing molecular epidemiological techniques for the study of C. daubneyi transmission patterns and prevalence in definitive and intermediate hosts. [Copyright &y& Elsevier]

A geographic information system (GIS) was constructed using remote sensing (RS) and landscape feature data together with Calicophoron daubneyi positive survey records from 197 georeferenced ovine farms with animals pasturing in a 3971 km2 area of the southern Italian Apennines. The objective was to study the spatial distribution of this rumen fluke, identify environmental features that influence its distribution, and develop a preliminary risk assessment model.The GIS for the study area was constructed utilizing the following environmental variables: normalized difference vegetation index (NDVI), land cover, elevation, slope, aspect, and total length of rivers. These variables were then calculated for “buffer zones” consisting of the areas included in a circle of 3 km diameter centered on 197 farms.The environmental data obtained from GIS and RS and from data taken by the veterinarians on the field (stocking rate and presence of streams, springs and brooks on pasture) were analyzed by univariate (Spearman and ANOVA) and multivariate (discriminant) statistical analyses using the farm coprological status (positive/negative) as the dependent variable.Sheep on 32 of the 197 (16.2%) farms, were positive for C. daubneyi, with an average intensity of 52 epg.A multivariate stepwise discriminant analysis model was developed that included moors and heathland, sclerophyllous and coniferous forest vegetation, autumn–winter NDVI and presence of streams, springs and brooks on pasture.The variables entered in the model were also correlated with C. daubneyi positive farms in the univariate tests and are consistent with the environmental requirements of C. daubneyi and its snail intermediate host. [Copyright &y& Elsevier]

• A 8-year integrated program to control CE was implemented in southern Italy. • New strategies have been developed and applied for animal-centered interventions. • The control programme resulted in a noteworthy reduction of CE in livestock. • The new strategies developed could be extended to other endemic Mediterranean areas. Cystic echinococcosis (CE) is a severe zoonosis, caused by the larval stage of the tapeworm Echinococcus granulosus. This helminth infection is of increasing public health and socio-economic concern due to the considerable morbidity rates that cause economic losses both in the public health sector and in the livestock industry. Control programmes against E. granulosus are considered long-term actions which require an integrated approach and high expenditure of time and financial resources. Since 2010, an integrated approach to control CE has been implemented in a highly endemic area of continental southern Italy (Campania region). Innovative procedures and tools have been developed and exploited during the control programme based on the following strategies: i) active and passive surveillance in livestock (using geospatial tools for georeferencing), ii) diagnosis in dogs (using the FLOTAC techniques and molecular analysis), iii) targeted treatment of farm dogs (using purpose-built confinement cages), iv) early diagnosis in livestock (by ultrasonography), v) surveillance in humans (through hospital discharge records analysis), vi) monitoring the food chain (analysing raw vegetables), vii) outreach activities to the general public (through dissemination material, e.g. brochures, gadgets, videos, virtual reality). Over eight years, the integrated approach and the new strategies developed have resulted in a noteworthy reduction of the parasite infection rates in livestock (e.g. up to 30 % in sheep). The results obtained so far highlight that using a one health multidisciplinary and multi-institution effort is of pivotal importance in preparing CE control programmes at regional level and could be extended to other endemic Mediterranean areas. [ABSTRACT FROM AUTHOR]