Calodium hepaticum (Nematoda: Capillaridae) in a red fox (Vulpes vulpes) in Italy with scanning electron microscopy of the eggs.

Calodium hepaticum (Bancroft, 1893) Moravec, 1982 (syn. Capillaria hepatica) is a cosmopolitan capillariid nematode, infecting mainly rodents and occasionally other mammals, including humans. Reports of C. hepaticum in canids are rare and the present one is, to the best of our knowledge, the first reported case in a red fox (Vulpes vulpes Linnaeus) in Italy. Scanning electron microscopy (SEM) examination of the eggs of C. hepaticum allowed a precise description of the egg morphology, which is one of the most relevant specific characteristics of capillariid nematodes. The egg shell showed a fibrous beam-like network which differs from that of the eggs of closely related trichinelloid species. This characteristic can be useful especially in case of spurious infection, when misdiagnosis among different trichinelloids species can occur.

Vulpes vulpes: a possible wild reservoir for zoonotic filariae.

Foxes (Vulpes vulpes, n = 132) killed during the hunting seasons 2005-2006 in Central Italy (Tuscany region) were examined in order to investigate the possible importance of this animal as a wild reservoir for zoonotic filariae. In each specimen adult worms of Dirofilaria immitis and hematic microfilariae were searched for. Species identification was performed by morphology, morphometry, the Barka staining technique applied to pulmonary and splenic blood smears, and, finally, by molecular diagnostics -- polymerase chain reaction (PCR) and sequencing. Twenty-three subjects (17.4%) proved to be positive for filarial parasites. Infection by Acanthocheilonema was more widespread than by Dirofilaria. Briefly, 8 foxes harbored mature adults of D. immitis; two of them (25%) also had microfilariae that in one case were mixed with the microfilariae of D. repens. Twelve subjects had microfilariae of Acanthocheilonema reconditum, and 3 harbored microfilariae of A. dracunculoides. Molecular diagnostics confirmed all results. Our findings, drawn by the examination of a few microliters of blood obtained from foxes approximately <2 years of age, support the hypothesis that this animal may be an abundant source of infection for ticks that transmit Acanthocheilonema parasites and for mosquitoes that act as vectors for dirofilarial nematodes. Therefore foxes, contributing to the parasite circulation in areas where dogs usually undergo prophylactic treatment, have to be considered an important wild reservoir for filarial parasites that can be transmitted to companion animals and people.

WWOX expression in different histologic types and subtypes of non-small cell lung cancer.

PURPOSE: Non-small cell lung cancer (NSCLC) has heterogeneous histopathologic classification and clinical behavior and very low survival rate. WWOX (WW domain-containing oxidoreductase) is a tumor suppressor gene, and its expression is altered in several cancers. The purpose of this study is to better define the role of WWOX in NSCLC tumorigenesis and progression by determining its pathogenetic and prognostic significance. EXPERIMENTAL DESIGN: WWOX protein expression was evaluated by immunohistochemistry in 170 patients with NSCLC (101 squamous cell carcinomas, 66 adenocarcinomas, 3 large cell carcinomas) and was correlated with histopathologic (histotype, subtype, grade, tumor-node-metastasis, stage, index of cell proliferation Ki67/MIB1) and clinical (age, gender, local recurrences, distant metastases, overall survival, and disease-free survival) characteristics. RESULTS: WWOX expression was absent/reduced in 84.9% of NSCLCs, whereas it was normal in 80.5% of adjacent normal lung tissues. WWOX expression was strongly associated with tumor histology (P=1.1x10(-5)) and histologic grade (P=0.0081): the percentage of cases with absent/strongly reduced WWOX expression was higher in squamous cell carcinomas and in poorly differentiated tumors. Regarding adenocarcinoma, bronchioloalveolar pattern showed normal WWOX expression in 62.5% of the cases, whereas in solid and acinar patterns, a prevalence of cases with absent/very low WWOX expression was observed (79.2% and 50%, respectively). Finally, weak WWOX staining intensity was related to the high index of cell proliferation (P=0.0012). CONCLUSIONS: Our results suggest that the loss of WWOX expression plays different roles in tumorigenesis of distinct histotypes and subtypes of NSCLC and is related to high aggressiveness (G3; high proliferating activity) of tumors.

Fishes and their parasites in the water district of Massaciuccoli (Tuscany, Central Italy).

This study has been conducted in the district of Massaciuccoli (lake, marsh and reclaimed areas with drainage channels) in Tuscany region (Central Italy). The aim of the research was to detect the presence of parasites in fishes, in particular of Opisthorchis felineus, which causes an important zoonosis. Between 2010-2012, the health status of 381 fishes was monitored, morphometric characteristics were determined, and parasites were searched for and identified. Of the 381 examined fishes, 189 were free of parasites while 192 were infected, among them 91 presented multiple infections. Opisthorchis felineus was not found in any of the examined fishes.

Applications of tissue microarray technology in immunohistochemistry: a study on c-kit expression in small cell lung cancer.

Tissue microarray technology allows the immediate evaluation of molecular profiles of numerous different tissues, with savings of money and time. It was created for rapid, large-scale molecular studies, and the main concern regarding its possible broad acceptance is that the analysis of tissue microarrays instead of whole tissue sections may lead to false negative or positive results because of tissue heterogeneity. In the present study, we analyzed in 54 small cell lung cancers, by immunohistochemistry, the expression of the antigen c-kit, which seems to be important in these neoplasms' tumorigenesis, and compared the staining obtained on whole sections with that of the corresponding tissue microarrays. Although c-kit expression of the whole sections agreed with that of the corresponding biopsies in many cases, the correlation between whole sections and all the companion nonlost single cores or their mean value turned out to be highly significant only if the 36 double negatives (ie, both whole sections and companion tissue microarrays negative) were included (P <0.0001). In fact, if only cases positive to at least 1 of the tests (i.e. whole sections or corresponding tissue microarrays positive) were considered, the correlation was not significant (P=0.055). Tissue microarrays showed a good specificity (94.2% for all single cores and 92.3% for their mean value) but a rather poor sensitivity (respectively, 69.4% and 71.4%). Moreover, a high percentage (13.4%) of cores was lost, and this loss was not random. To sum up, in our experience, tissue microarray technology cannot be a substitute for whole sections in clinical diagnosis of individual cases.

West Nile seroprevalence study in Bolivian horses, 2011.

West Nile virus (WNV) is a mosquito-borne virus belonging to the family Flaviviridae included in the Japanese encephalitis antigenic complex (JEAC). A seroepidemiological study was carried out in 2011 using 160 horse sera collected from different areas of Bolivia to investigate the presence of WNV antibody. A high proportion (59.4%) of the tested sera were positive to a commercially available WNV competitive enzyme-linked immunosorbent assay (C-ELISA). Sixty-six randomly selected C-ELISA-positive sera were further tested by WNV plaque reduction neutralization test (PRNT), virus neutralization (VN), and immunoglobulin M (IgM)-WNV ELISA to exclude false-positive results due to possible cross-reactions to other members of the JEAC and to investigate if the horses were recently infected. No WNV IgM was detected in these samples, whereas neutralizing antibodies were found in 21 and 18 samples by PRNT and VN, respectively. In conclusion, a high proportion of the Bolivian horses included in this study reacted serologically against viruses of the JEAC. WNV was partially responsible (31.8%) for these reactions, supporting the conclusion that WNV circulated in Bolivia.

Expression profiling of angiogenic genes for the characterisation of colorectal carcinoma.

The development of new blood and lymphatic vessels is a crucial event for cancer growth, metastatic spread and relapse after therapy. In this work, the expression levels of chemokines, angiogenic and angiostatic factors and their receptors were determined in paired mucosal and tumour samples of patients with colorectal carcinoma and correlated with clinical and histological parameters by advanced multivariate analyses. The most important predictors to discriminate between tumour and paired normal mucosa turned out to be the levels of expression of plexin-A1 and stromal cell-derived factor 1 (SDF-1), the former overexpressed and the latter downregulated in tumours. The levels of osteopontin and Tie-2 transcripts discriminated between the presence and absence of lymph node infiltration, the former overexpressed in the presence of infiltration whilst the latter providing a protective role. These results add support to the notion that the expression levels of selected genes involved in new blood and lymphatic vessel formation represent trustable biomarkers of tumour development and invasion and contribute to the identification of novel molecular classifiers for colorectal carcinoma.

Different estrogen receptor beta expression in distinct histologic subtypes of lung adenocarcinoma.

Adenocarcinoma is becoming the most common histologic type of lung cancer in both sex. Although most cases are seen in smokers, it develops more frequently than other histologic types in individuals who have never smoked. This evidence suggests that other putative etiologic factors, such as sex hormones, need to be investigated. Several subtypes of lung adenocarcinoma have been recently described with distinct clinicopathologic features and prognostic implications. The purpose of this study is to investigate the role of estrogen receptor beta in lung adenocarcinoma, with particular attention paid to its different histologic subtypes. Nuclear estrogen receptor beta expression was evaluated by immunohistochemistry in 112 lung adenocarcinomas, including both "single subtype" and "mixed subtype" samples. Using a 2-level (high/low) score system, estrogen receptor beta expression was high in most (75%) adenocarcinomas and turned out to be strongly related to the histologic subtypes. In fact, estrogen receptor beta expression was low or negative in 68.2% of solid subtypes, whereas it was high in 76.5% of nonmucinous bronchioloalveolar, in 69.4% of acinar, and in 61.2% of papillary patterns (P = .00004). Furthermore, a strong association between estrogen receptor beta expression and tumor histologic grade was observed: estrogen receptor beta was highly expressed predominantly in well- and moderately differentiated tumors (P = .0014). In conclusion, estrogen receptor beta expression has distinct patterns in lung adenocarcinoma, suggesting a specific role for estrogen receptor beta in the pathogenesis of different histologic subtypes of this type of cancer. Moreover, loss of estrogen receptor beta expression in poorly differentiated (G3) tumors could represent a crucial step in the dedifferentiation process of lung adenocarcinoma.