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Biomedical subjects

Fabio Del Piero

Publications and source records attributed to Fabio Del Piero.

10 recordsLinked to original sources

The neutralizing antibody response against West Nile virus in naturally infected horses.

A major neutralizing epitope (here referred to as the T332 epitope) located on the lateral surface of domain III (DIII) of the West Nile virus (WNV) envelope protein has been identified based on the analysis of murine monoclonal antibodies. However, little is known about the humoral immune response against WNV in a natural host or whether DIII in general or the T332 epitope in particular are important targets of neutralizing antibodies in vivo. To characterize the types of antibodies produced during infection with WNV, we studied a group of naturally infected horses. Using immune adsorption assays coupled with the use of virus particles bearing mutations in the T332 epitope, we found that in some animals neutralizing activity against DIII and the T332 epitope was below the limit of detection. In contrast, some animals generated a significant fraction of neutralizing activity to DIII and the T332 epitope. Thus, while antibodies to the T332 epitope did not represent a significant fraction of the total antibody response in the infected animals studied, in some horses, they comprised a significant fraction of neutralizing activity, making this an important but far from dominant neutralizing epitope. Rather, the neutralizing response to WNV generated in infected horses is both variable and polyclonal in nature, with epitopes within and outside of DIII playing important roles.

Animals↗

Chlorophacinone exposure causing an epizootic of acute fatal hemorrhage in lambs.

This report describes an epizootic of chlorophacinone toxicosis in lambs with severe acute hemorrhages. Eleven lambs, approximately 1-2 months of age, suddenly developed epistaxis, respiratory distress, and facial and cervical swelling. Affected animals died within 1-2 hours from the onset of clinical signs. Two lambs were available for complete postmortem examination. Gross lesions included mucosal and organ pallor, icterus, melena, and lung edema, as well as thymic, cervical muscle, and intra-articular hemorrhage. Histologically hepatocellular centrolobular necrosis was observed. The anticoagulant chlorophacinone was detected in the livers at 0.58 ppm and 0.50 ppm (wet weight), respectively. The source of exposure to chlorophacinone was old bait material placed between the wall studs of the building housing the ewes and lambs. The lambs were able to reach the bait through a hole in the plywood interior wall of the building.

Animals↗

Necropsy findings in American alligator late-stage embryos and hatchlings from northcentral Florida lakes contaminated with organochlorine pesticides.

Increased American alligator (Alligator mississippiensis) embryo and neonatal mortality has been reported from several northcentral Florida lakes contaminated with old-use organochlorine pesticides (OCPs). However, a clear relationship among these contaminants and egg viability has not been established, suggesting the involvement of additional factors in these mortalities. Thus, the main objective of this study was to determine the ultimate cause of mortality of American alligator late-stage embryos and hatchlings through the conduction of detailed pathological examinations, and to evaluate better the role of OCPs in these mortalities. Between 2000 and 2001, 236 dead alligators were necropsied at or near hatching (after approximately 65 days of artificial incubation and up to 1 mo of age posthatch). Dead animals were collected from 18 clutches ranging in viability from 0% to 95%. Total OCP concentrations in yolk ranged from approximately 100 to 52,000 microg/kg, wet weight. The most common gross findings were generalized edema (34%) and organ hyperemia (29%), followed by severe emaciation (14%) and gross deformities (3%). Histopathologic examination revealed lesions in 35% of the animals, with over half of the cases being pneumonia, pulmonary edema, and atelectasis. Within and across clutches, dead embryos and hatchlings compared with their live cohorts were significantly smaller and lighter. Although alterations in growth and development were not related to yolk OCPs, there was an increase in prevalence of histologic lesions in clutches with high OCPs. Overall, these results indicate that general growth retardation and respiratory abnormalities were a major contributing factor in observed mortalities and that contaminants may increase the susceptibility of animals to developing certain pathologic conditions.

Alligators and Crocodiles↗

Indirect immunohistochemistry on skin biopsy for the detection of persistently infected cattle with bovine viral diarrhoea virus in Italian dairy herds.

Indirect immunohistochemistry (IHC) on skin biopsies for identification of persistently infected (PI) animals has been used as a parallel test to antigen and antibody ELISAs in a bovine viral diarrhoea (BVD) voluntary control program. The aim was to evaluate the reliability and feasibility of IHC on ear skin tissues to detect PI animals in field conditions, including both adult and calves under 6 months of age. In animals over 6 months of age skin biopsy and blood sample were collected at the same time, whereas in young calves blood sampling was performed when animals reached 6 months of age. One hundred and sixty-five animals were tested and immunohistochemical results were compared with those of antigen ELISA. In case of inconclusive results virus isolation and virus neutralization assays were performed. Agreement K value was 0.96. Immunohistochemical staining in positive animals was clearly detectable in the keratinocytes of the epidermis and adnexa.

Animals↗

Histone H1 proteins act as receptors for the 987P fimbriae of enterotoxigenic Escherichia coli.

The tip adhesin FasG of the 987P fimbriae of enterotoxigenic Escherichia coli mediates two distinct adhesive interactions with brush border molecules of the intestinal epithelial cells of neonatal piglets. First, FasG attaches strongly to sulfatide with hydroxylated fatty acyl chains. This interaction involves lysine 117 and other lysine residues of FasG. Second, FasG recognizes specific intestinal brush border proteins that migrate on a sodium-dodecyl sulfate-polyacrylamide gel like a distinct set of 32-35-kDa proteins, as shown by ligand blotting assays. The protein sequence of high performance liquid chromatography-purified tryptic fragments of the major protein band matched sequences of human and murine histone H1 proteins. Porcine histone H1 proteins isolated from piglet intestinal epithelial cells demonstrated the same SDS-PAGE migration pattern and 987P binding properties as the 987P-specific protein receptors from porcine intestinal brush borders. Binding was dose-dependent and shown to be specific in adhesion inhibition and gel migration shift assays. Moreover, mapping of the histone H1 binding domain suggested that it is located in their lysine-rich C-terminal domains. Histone H1 molecules were visualized on the microvilli of intestinal epithelial cells by immunohistochemistry and electron microscopy. Taken together these results indicated that the intestinal protein receptors for 987P are histone H1 proteins. It is suggested that histones are released into the intestinal lumen by the high turnover of the intestinal epithelium. Their strong cationic properties can explain their association with the negatively charged brush border surfaces. There, the histone H1 molecules stabilize the sulfatide-fimbriae interaction by simultaneously binding to the membrane and to 987P.

Adhesins, Escherichia coli↗

Correlation of in vivo photosensitizer fluorescence and photodynamic-therapy-induced depth of necrosis in a murine tumor model.

We compared light-induced fluorescence (LIF) to nominal injected drug dose for predicting the depth of necrosis response to photodynamic therapy (PDT) in a murine tumor model. Mice were implanted with radiation-induced fibrosarcoma (RIF) and were injected with 0, 5, or 10 mg/kg Photofrin. 630-nm light (30 J/cm(2), 75 mW/cm(2)) was delivered to the tumor after 24 hours. Fluorescence emission (lambda(excitation)=545 nm, lambda( emission)=628 nm) from the tumor was measured. The LIF data had less scatter than injected drug dose, and was found to be at least as good as an injected drug dose for predicting the depth of necrosis after PDT. Our observations provide further evidence that fluorescence spectroscopy can be used to quantify tissue photosensitizer uptake and to predict PDT tissue damage.

Animals↗

Photodynamic therapy creates fluence rate-dependent gradients in the intratumoral spatial distribution of oxygen.

In photodynamic therapy (PDT), treatment efficacy may be reduced by the presence of pre-existing tumor hypoxia or by oxygen depletion during the therapy. Tumor oxygenation during PDT has been measured with needle electrodes, but the intratumoral distribution of this oxygen is not known. In the present study, the spatial distribution of hypoxia during PDT was quantified using the hypoxia-labeling marker EF3. Mice bearing radiation-induced fibrosarcoma tumors were treated with Photofrin-mediated PDT to a total dose of 135 J/cm(2), delivered at a fluence rate of either 75 mW/cm(2) or 38 mW/cm(2). PDT-created hypoxia at each fluence rate was labeled by exposing tumors to EF3 (52 mg/kg) during the period of illumination. Cryosectioning, immunohistochemistry, and fluorescence microscopy were carried out to quantify EF3 binding as a function of distance to the nearest perfused blood vessels in sections cut from within the superficial (light-adjacent) 600 micro m or the deep (light-distant) 600 micro m of tumors (5-6 mm in diameter, approximately 3 mm in depth). In both superficial and deep sections, PDT at 75 mW/cm(2) resulted in the development of significant gradients in tumor hypoxia as a function of distance to a perfused blood vessel. Furthermore, significant hypoxia was detected even in vascular-adjacent tissue. These effects were associated with a significant decrease in the percentage of perfused vessels and a significant increase in the median distance of a cell to the nearest perfused blood vessel. In contrast, during PDT at 38 mW/cm(2), sections from deep tumor levels demonstrated only insignificant increases in the rise in hypoxia as a function of distance to a perfused vessel and in the level of hypoxia in vascular-adjacent tissue. No effects on tumor perfusion were detected during PDT at 38 mW/cm(2). Overall, these results demonstrate that spatially dependent depletion of oxygen can occur during PDT as a function of the fluence rate and that PDT can create significant hypoxia in even tissue adjacent to perfused blood vessels.

Animals↗

Metastatic melanoma in horses.

The clinical and pathologic findings are reviewed for 14 horses with metastatic melanoma. All were older gray horses, with an average age of 16 years. The most common sites of primary tumors were the ventral tail, perineum, and parotid salivary gland. Metastases were found in multiple locations and caused a variety of clinical syndromes. The most common sites for metastases were the lymph nodes, liver, spleen, skeletal muscle, lungs, and surrounding or within blood vessels throughout the body. Many of the horses had dermal melanomas for years (range 1-6 years) before succumbing to metastatic disease. Histologic characteristic of dermal masses was not predictive of malignancy in the majority of cases. Treatment consisting of surgical debulking and administration of cimetidine, an autogenous vaccine, or both was attempted in 4 horses with no effect on outcome.

Adjuvants, Immunologic↗

Hemangiosarcoma in 11 young horses.

Hemangiosarcoma is a rare neoplasm of horses and hemangiosarcoma in young horses might behave differently than in mature horses. The purpose of this study was to identify the characteristics of hemangiosarcoma occurring in horses < or = 3 years of age. Medical records from 1982 to 2004 were searched for horses < or = 3 years of age with a histopathologic diagnosis of hemangiosarcoma. Eleven records were identified. Thoroughbred and Thoroughbred crosses predominated. Age ranged from 9 days to 3 years. All horses presented with cutaneous or leg swellings or joint effusion. Physical examination findings included tachycardia, fever, and depression. Laboratory abnormalities included anemia (5/11), hyperfibrinogenemia (4/11), hypofibrinogenemia (3/11), thrombocytopenia (2/11), and neutrophilic leukocytosis (1/11). Ultrasonographic and radiographic evaluation was not diagnostic in any case. Antemortem histopathologic diagnosis was obtained in 10 cases. Six of 11 horses were euthanized. Surgical resection was performed in 5 horses, 2 of which were later euthanized. Diagnosis was confirmed histologically at postmortem examination in all euthanized horses. Two cases resolved spontaneously. Early histopathologic diagnosis may allow cure if the mass is localized and amenable to surgical resection. In cases where the horse is medically stable, and masses are not interfering with quality of life, a period of observation may be warranted.

Age Factors↗