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Biliary tract brush cytology is increasingly being recognized as a favoured method for evaluating abnormalities of the biliary tract. In order to increase the diagnostic accuracy, we devise a new brush processing method finalized to the complete and ideal cytologic examination of the collected material. Small fragments of the mucosa, of inflammatory cell aggregates or of carcinomas are observed and the results are optimally fixed and allow a definitive histological diagnosis.

Although endoscopic ultrasound combined with fine needle aspiration (EUS-FNA) is rapidly becoming the preferred diagnostic approach for the sampling and diagnosis of gastrointestinal and mediastinal malignancies, there are limited data as to its use in the diagnosis of lymphoproliferative disorders. Therefore, we carried out a retrospective evaluation of the performance of EUS-guided FNA combined with flow cytometry (FC) as a tool to improve overall sensitivity and specificity in the diagnosis of lymphoma.Of 1560 patients having EUS-guided FNA during the period of the study, a total of 56 patients were evaluated by cytology with FC after EUS-FNA. There was adequate material to perform FC analysis for all but one case.EUS-FNA-FC gave a diagnosis of lymphoma in 11 cases and of reactive lymphadenopathy in 20. A specific histological type was defined by FC alone in eight cases. The remaining cases were diagnosed later by cytology and cell block sections: 13 carcinomas, nine granulomatous lymphadenopathies and one mediastinal extramedullary haematopoiesis. One case was considered only suspicious for lymphoma on cytology and FC but was not confirmed on molecular analysis and one had insufficient material for FC.Our results show that a combination of EUS-FNA-FC is a feasible and highly accurate method, which may be used for the diagnosis and subtyping of deep-seated lymphoma, providing a significant improvement to cytomorphology alone both for diagnosis and treatment planning, as long as immunocytochemistry is available for non-lymphoma cases.

Introduzione. La citologia su striscio delle vie biliari extraepatiche effettuata con spazzolino (brushing endoscopico) è il metodo più utilizzato per la valutazione delle lesioni delle vie biliari extraepatiche, ma presenta scarsa sensibilità. Per migliorarne la potenzialità diagnostica, presentiamo una nuova ed originale metodica che permette di effettuare un esame citologico ottimale del materiale ago-aspirato. Metodi. Lo spazzolino viene immediatamente immerso in metanolo e inserito nelle biocassette per l’inclusione in paraffina. Vengono tagliate delle sezioni parallele e consecutive lungo l’asse maggiore, fino al core metallico; successivamente il blocco di paraffina viene ruotato di 180° e nuove sezioni vengono effettuate sul lato opposto. Le sezioni, colorate in Ematossilina e Eosina e con Alcian Blue Mucine, hanno mostrato piccoli frammenti di mucosa, di cellule infiammatorie aggregate o di carcinoma, con un’ottima fissazione, permettendo una diagnosi definitiva accurata in quasi la totalità dei casi esaminati. Infatti 112 campioni citologici di brushing endoscopici delle vie biliari extraepatiche (67 M; 45 F), raccolti all’Ospedale “Molinette”, Università di Torino, tra Gennaio 2002 e Agosto 2006, sono stati inclusi nel nostro studio. Tutti i pazienti avevano una diagnosi definitiva istologica e un follow-up clinico medio di 21 mesi. Risultati. Confrontando questa originale metodica su 112 casi consecutivi di brushing endoscopico con la diagnosi istologica su pezzo operatorio, abbiamo osservato: 87% di sensibilità, 100% di specificità, 100% di valore predittivo positivo (VPP) e 91% di valore predittivo negative (NPV). La metodica inoltre, confrontata con la diagnosi clinica dopo almeno 6 mesi di follow-up, ha mostrato: 88% sensibilità, 100% specificità, 100% VPP e 96% VPN. Conclusioni. In conclusione questa nuova metodica è altamente sensibile e specifica, limitando all’1% le diagnosi non idonee, con una concordanza con la diagnosi istologica pari all’88% (K-value).

The authors describe a case of acute diverticulitis of the appendix presenting as an inflammatory mass in the right lower quadrant of the abdomen. The histologic and clinical findings are discussed. This uncommon condition may mimic acute appendicitis, but in most instances it departs from typical appendicitis for later age of appearance, more indolent clinical course and increased tendency to perforation. In these cases the disease may progress with subacute interstitial inflammation, with or without abscess formation, and present as a tumor-like mass of the cecum.

Fine needle aspiration (FNA) of the thyroid, which is a rapid and cost-effective procedure, has gained wide acceptance as a valuable method for distinguishing neoplastic from non-neoplastic nodules and identifying those patients requiring surgery 1-4. At the present time, the thyroid gland is the most frequently aspirated organ for triage/diagnostic purposes. Clinically-relevant thyroid nodules occur in 5-10% of the general population in Italy and approximately 5% of the patients have malignant lesions. The goal of thyroid FNA is to identify the nodules that require surgery and decrease the number of thyroidectomy for patients with benign disease. Overall, the technique has a high sensitivity and specificity for the detection of thyroid neoplasm 5-7. Cell blocks (CBs) are often prepared on FNAs from several organ sites as an adjunct to smears in the diagnosis of potential lesions. However, the literature contains few reports on their utility with regard to specific organ sites. The main advantage of the CB is the potential to produce several sections for special stains and other ancillary, in particular immunoistochemical, studies 8-10. At our institution, CBs are made routinely on thyroid FNAs since twenty years and we have been performing about eight-hundred thyroid FNAs each year. As a result, we have been collecting a high number of CB of thyroid lesions, of great value for investigative and retrospective studies. The aspirates were procured by experienced cytopathologists, clinicians, or radiologists. The majority of the aspirates were performed by the radiologist or clinicians under ultrasound guidance and each aspirate had an immediate in situ assessment for adequacy by the cytopathologist. FNAs were performed using a 23- or 25-gauge needle attached to a 10-ml disposable syringe mounted to a metallic Cameco syringe pistol (Morton Medical Ltd., London, United Kingdom). One to 4 aspirates were performed per case. After each sampling, air-dried Hematoxylin & Eosin-stained slides were made for immediate assessment and alcohol-fixed slides for subsequent Papanicolaou (Pap) staining. The residual material and the needles were rinsed immediately in a 50% Ethanol solution (s.c. Lysis Buffer) containing Ammonia Chloride, Potassium Bicarbonate and EDTA, which proved effective for fixing cells and lysing erythrocytes. Material collected by centrifugation was embedded in paraffin.presence of cellular fragments. CB has greatly improved the pre-surgical diagnosis of thyroid nodules, since small tissue specimens representative of the lesion were detected in the great majority of cases. These “micro-biopsies” provided morphologically-relevant data. Besides, sections were employed for immunoistochemical and molecular tests. Expression of galectin-3 is routinely tested on thyroid follicular lesions (cytological cell-blocks) obtained preoperatively by ultrasound-guided fine-needle aspiration of thyroid nodules 11. Moreover, CB procedure provides the basis for new molecular and immunoistochemical studies to determine the definition of thyroid neoplasm of indeterminate malignant behavior (Thy3). Recently, we focused on the immunohistochemistry of Emerin, a protein of the nuclear membrane (NM) whose decoration best demarcates the nuclear shape of the thyrocytes 12 and we performed this stain on a series of 82 cytological CB thyroid specimens. Emerin revealed a uniform arrangement of the NM in non-neoplastic lesions (thyroiditis, microfollicular goiter, follicular adenoma) and normal thyroid as well as in follicular carcinoma. In contrast, irregular folding of the membrane and presence of curling and invaginations, eventually leading to the formation of nuclear pseudoinclusions, was observed in PTC and VFPTC cells. In conclusions, the integration of CB method with conventional cytomorphological and clinical diagnostic procedures represents a sensitive and reliable diagnostic approach for preoperative identification of thyroid carcinomas. This procedure improves the diagnostic accuracy of conventional cytology. Moreover, the CB is useful for immunoistochemical and molecular study of thyroid neoplasm

Preoperative CA 125 levels were measured in 36 patients with advanced epithelial ovarian carcinoma in clinical response undergoing a second-look operation. All the patients had positive levels (> 35 U/ml) of this tumor marker at diagnosis. The correlation between antigen levels and disease status at surgery revealed a sensitivity of this assay of 0.55 (only 11/20 patients still with tumor had positive levels) and a specificity of 0.94 (15/16 patients with no tumor had < 35 U/ml). The predictive value of a positive test was 0.92. This method unfortunately proved unable to recognize microscopic residual tumor burden, less than 0.5 cm.

Objectives. Endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) is increasingly used in diagnosis of pancreatic tumors. Our study aims to identify cytological characteristics of on-site evaluation helpful for recognizing pancreatic neuroendocrine tumors (NETs). Methods. Our study comprises 5 cases of pancreatic tumors with cytological features suggestive of neuroendocrine origin, obtained by EUS-FNA which diagnosis was confirmed by surgical excision. For each case we analized: cytological smears, cell block sections and immunocytochemical stains. Results. All cases show similar and peculiar cytological characteristics of on-site evaluation: smears were richly cellular with poorly cohesive small cells; they display a monotonous basophilic appearance with scanty cytoplasm and small eccentric nucleus with speckled or dusty chromatin pattern conferring a plasmocytoid appearance. Immunocytochemical stains (Chromogranin A, Synaptophysin), performed on cell-block, confirmed neuroendocrine differentiation. Conclusion. EUS-FNA is a useful method of identification of pancreatic NETs. Recognizing cytopathological features of neuroendocrine origin improves the diagnostic yield of the procedure, allowing a three-tiered approach in planning the handling of FNA specimens.

Bacillus thuringiensis (Bt) is a ubiquitous bacteria widely used as a biopesticide to control a number of important insect pests. Since the mid-1990s, genetically modified (GM) plants expressing Bt genes have been used as an effective tool to control a wide range of insect pests. In recent years, a wide number of articles addressing the environmental impact of genetically modified plants have been published. However, only a few have addressed the occurrence and distribution of the indigenous Bt population in agricultural systems. Here, culturing and molecular methods were used to study the occurrence of Bt harboring insecticidal cry1 genes in a corn field. Samples of corn leaves and soil were collected in July and August 2007 from a 10 ha corn field in Northern Italy. The results showed that the highest Bt density was in leaves located near the soil surface. The incidence of Bt isolates harboring antilepidopteran cry1 genes was 42% of the total tested isolates. Approximately 20% of the Bt isolates harbored the cry1Ab and cry1Ac genes. Similarly to Bt density, the highest abundance of isolates harboring cry1 genes was found in leaves collected near the plant collar. Less than 9% of the Bt isolated from soil harbored cry1 genes. Density of Bt was reduced by application of the insecticide chlorpiryfos. This effect appeared to be due to simply washing off effects of the insecticide treatment to the corn phyllosphere. This study showed that Bt was fairly abundant in the corn agroecosystem and that the high incidence of isolates harboring antilepidopterean cry1 genes could have a role in preserving the sustainability of the agroecosystem.

AIM: To increase the diagnostic potential of endoscopic biliary tract brushing, we devised an approach alternative to cytological smearing, leading to the preoperative histological examination of the collected material. METHODS: One hundred twelve consecutive biliary brush specimens were included. All patients presented a stricture of the biliary tract, leading to a diagnostic procedure by brushing. Immediately following brushing, the endoscopist immersed the brush into methanol and sent it to the pathology laboratory. The brush was introduced into a cassette for paraffin embedding and sections parallel to the long axis of brush were cut until the metal wire was almost reached, then the block was rotated and new sections were obtained from the opposite side. Samples of the mucosa, inflammatory cell aggregates, small fragments of carcinomas, or isolated cells were observed, and displayed an optimal fixation, allowing a definite diagnosis that proved mandatory for therapy in the vast majority of cases (99.1%). RESULTS: The results obtained in 112 consecutive cases using such technique compared with final histological diagnosis proved: 91% sensitivity, 100% specificity, 100% positive predictive value (PPV), and 87% negative predictive value (NPV) (P < 0.001). In nonoperated patients, the clinical diagnosis after at least 6 months of follow-up showed: 95.5% sensitivity, 100% specificity, 100% PPV, and 88.2% NPV (P < 0.001). CONCLUSION: Such novel approach to the preoperative diagnosis of biliary tract lesions proved to be highly sensitive and specific, limiting the inadequate preoperative diagnoses to less than 1%.

A semi-field experiment was carried out in two peach orchards in northern Italy to assess mortality due to predators and parasitoids on the exotic coccinellid Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) in comparison with the native coccinellid Adalia bipunctata L. (Coleoptera: Coccinellidae). The experiments were conducted in cages to avoid the possible escape of the exotic ladybird (not yet established in Italy). Two kinds of cage experiments were included: ‘exclusion cages’ (access by walking predators impeded) and ‘free cages’ (walking predators free to enter). The cages, containing all the stages of the two ladybird species, were placed in two localities and left for 24 h. All ladybird stages used for the semi-field experiments came from a laboratory rearing. The eggs of H. axyridis experienced less mortality than those of A. bipunctata. The ant workers were the most frequent predators in ‘free cages’ but A. bipunctata cannibalism on eggs was also detected. Larvae of both coccinellid species were predated equally but larval predation of L1 and L2 was higher in comparison to predation of L3 and L4. Pupae and adults of both exotic and native ladybirds were never attacked by predators. Predation on younger larval stages was higher in the ‘free cages’ in comparison with ‘exclusion cages’. No ladybird parasitisation was observed. The ‘free cage’ technique seems to provide a standardised and realistic estimation of predation impact but more studies are needed to evaluate ladybird parasitisation in semi-field conditions.

As consequence of the concern about the biosafety of genetically modified plants, biological and ecological studies are considered crucial for environmental risk assessment. Laboratory experiments were carried out in order to evaluate the transfer of the Cry1AcBt-toxin from a transgenicBt-oilseed rape to a non-target pest,Myzus persicaeSulzer. Cry1Ac protein levels in plants and aphids were determined using a double sandwich enzyme-linked immunosorbent assay. Phloem sap from (Bt+) and (Bt−) oilseed rape plants was collected from leaves using a standard method of extraction in an EDTA buffer.Bt-toxin was present in phloem sap, with a mean concentration of 2.7±1.46 ppb, corresponding to a 24-fold lower level than in oilseed rape leaves. Toxin was also detected in aphid samples, with a mean concentration in the positive samples of 2.0±0.8 ppb. The evidence thatBt-toxin remains in herbivores, in this case an aphid, could be useful to clarify functional aspects linked to possible consequences ofBt-crops on food chains involving herbivore–natural enemy trophic systems. Further studies are needed in order to improve the knowledge on the functional aspects linked to the transfer of the Cry1AcBt-toxin from GM-oilseed rape to aphids and their possible consequence.

As part of an environmental risk assessment study of exotic natural enemies used in inundative biological control, life-history characteristics of Harmonia axyridis (Pallas), Hippodamia variegata (Goeze) and Adalia bipunctata (L.) (Col., Coccinellidae) were quantified under laboratory conditions at 25°C on Myzus persicae (Sulzer) as prey. Comparative studies showed significant differences among pre-adult development times: H. axyridis developed slower ( = 19.8 days) than H. variegata ( = 18.1 days) and A. bipunctata ( = 18.4 days). Differences were also evident in the duration of egg, larval and pupal stages. No measurable differences among the three species were found for fecundity, oviposition rate and adult longevity. Harmonia axyridis exhibited the longest pre-oviposition ( = 7.4 days) and interoviposition ( = 3.6 days) periods and the shortest oviposition period ( = 13.7 days). The Bieri model was used to describe age-specific fecundity for the three species of coccinellids. The intrinsic rate of increase (rm), net reproductive rate (R0) and mean generation time (T) were higher for H. variegata (rm = 0.114, R0 = 52.75, T = 41.88 days) than for H. axyridis (rm = 0.089, R0 = 26.27, T = 38.81 days) or A. bipunctata (rm = 0.081, R0 = 18.49, T = 40.06 days). Our findings show that the biological traits of H. axyridis do not seem to be factors that may contribute to the invasiveness of this coccinellid.