Hydrated nucleus pulposus herniation in seven dogs.

The clinical signs, magnetic resonance imaging (MRI) findings, treatment and follow-up in seven dogs with hydrated nucleus pulposus extrusion (HNPE) are reported. All dogs had tetraparesis or tetraplegia. T2-weighted MRI revealed extradural hyperintense homogeneous material compressing the cervical spinal cord. After conservative treatment (five dogs) or surgical decompression (two dogs), all dogs returned to ambulatory function within 1 month. Follow-up MRI in conservatively treated dogs revealed complete disappearance of the extruded material. Histopathological examination of surgical specimens confirmed that the retrieved material was extruded nucleus pulposus with evidence of early degeneration.

New insights onto cardiopulmonary nematodes of dogs in Sardinia, Italy.

Dog heartworms Angiostrongylus vasorum and Dirofilaria immitis cause severe parasitological diseases; the importance of these parasitosis is growing due to their health impact on animals, the possible zoonotic implications and the recent spreading across several European countries and previously non-endemic areas. The aim of this study is to update the epidemiological scenario of cardiopulmonary nematodes A. vasorum and D. immitis in dogs of Sardinia island and to perform a morphological identification of larvae by the use of the Baermann and Knott techniques respectively and the molecular characterization of the mitochondrial cytochrome c oxidase subunit I (cox1) and the second ribosomal transcribed spacer region (ITS-2) of larvae L1 of A. vasorum. In the present study, 3.4% (5/146) of dogs resulted positive at Baermann technique for A. vasorum while 8.9% (61/684) to D. immitis. If on one side A. vasorum can be considered an emerging parasite in Sardinia, the parasitic pressure and the risk of infection for D. immitis in the island seems to be increased compared with the recent past.

Molecular characterization of subcutaneous and muscular coenurosis of goats in United Arab Emirates.

Coenurosis is a parasitic disease caused by the larval stage of Taenia multiceps, which affects various ruminants species, particularly sheep and goats, and occasionally humans. In this note, non-cerebral coenurosis in goats with the goal to compare morphological and biomolecular characteristics is focused in order to determine if the non-cerebral specimens should be considered a new specie or a strain of T. multiceps. Three hundred goats slaughtered with macroscopic lesions due to the presence of metacestodes were examined in an abattoir in Dubai, United Arab Emirates in order to evaluate the presence of extra-cerebral coenuri. Forty-eight coenuri were found under the skin, between fasciae of the skeletal muscles, diaphragm and in the abdominal cavities. Morphologically, the examined non-cerebral coenuri recovered showed the same features reported by other authors for Coenurus cerebralis, but their location outside the central nervous system suggests that they may be a different strain or genetic variants of T. multiceps. Nine caprine coenuri were then processed for sequencing of mitochondrial partial COI (396 bp) and ND1 (471 bp) genes indicating that they had a pairwise distance of 1.0-1.3% and 2.4-4.1% compared with parasite' COI sequences from Italy (Tm1-Tm3 strains) and Erzurum strains of ovine origin, respectively; whereas it had 0.6-1.3% and 0.4-1.1% pairwise distance for ND1. Phylogenetic trees of their ND1 and COI sequences using the maximum composite likelihood method with MEGA showed that these nine parasites recovered in Emirates abattoir could be grouped into a new strain. In conclusion, morphological and molecular characterization of caprine coenurosis from Dubai, United Arab Emirates, shows that the specimens are significantly different from other strains or genotypes in COI sequence, which suggests that they should belong to different genotypes or strains of T. multiceps.

Lumbar interbody expanding cage. A preliminary study on an animal model.

Interbody fusion devices are used in human medicine for treating degenerative diseases of the spine. Currently, there is not a universally accepted assessment tool for determining fusion, and the definitive criteria for diagnosing a successful interbody fusion remain controversial. The aim of this study was to describe microscopic and helical computed tomography (CT) imaging in the assessment of lumbar interbody fusion using cylindrical threaded titanium expanding cage in sheep. One cylindrical threaded expanding titanium cage (Proconcept--SA, Orange, France) was inserted through a transperitoneal approach after radical discectomy and packed with cancellous bone autograft in five adult sheep. The subjects were euthanatized after three, six, 12, 18 and 24 months. CT images revealed lumbar fusion at 12 months post operation, whereas microscopic evaluations indicated the presence of lumbar fusion at 18 months. CT and histological grades were the same in 65% of the cases observed. There were not a significant difference between CT, histological and micro radiographic grades. Helical CT scanning can be considered to be a suitable method for the monitoring of lumbar fusion as it enables observation of the deposition of bony bridging within the cage.

Nodular lesions of the tongue in canine leishmaniosis.

In this case report, an atypical clinical presentation of leishmaniosis in a dog with multiple nodular lesions of the tongue is described. Haematological and biochemical analysis, serological test for Leishmania infantum antibodies and biopsy samples from several nodules of the tongue for histopathological examination were made. The final diagnosis of leishmaniosis was based upon the observation of amastigotes in the bioptic samples. It is recommended to consider leishmaniosis among the list of differentials of mucosal nodular lesions, at least in endemic areas.

Carboplatin sustained delivery system using injectable microspheres.

A controlled carboplatin delivery system using biodegradable polymer has been used in this study. The purpose was to evaluate the local and systemic effects of injectable, biodegradable microspheres containing carboplatin when injected as aqueous suspension subcutaneously in rats. Biocompatibility and toxicity of empty microspheres and microspheres loaded with carboplatin were evaluated by clinical and histological examination. The diffusion of carboplatin in tissues and time of drug release were evaluated by platinum determination in plasma and tissues over the time. The results of the study suggest that microspheres provide a sustained slow release of carboplatin and that multiple inoculations of microspheres containing drug and no evidence of local or systemic toxicity is found. This device may be useful in the treatment of solid tumours.

Phenotypic and functional differences between human saphenous vein (HSVEC) and umbilical vein (HUVEC) endothelial cells.

The vascular endothelial cell (EC) plays an essential role in the pathogenesis of inflammation, transplant rejection and tumour metastasis. Most research on vascular ECs uses human umbilical vein endothelial cells (HUVECs). However, HUVECs are derived from immune-naive foetal tissue, and show significant functional differences from adult vascular endothelium. In this paper, we characterise an alternative model based on human saphenous vein ECs (HSVECs), describe their culture conditions and provide a detailed functional comparison with HUVECs. Compared with HUVECs, HSVECs show an increased sensitivity to ox-LDL and a reduced response to cytokines, as indicated by adhesion molecule expression as well as leukocyte adhesion and transmigration. With respect to their ability to present antigen, HSVECs have a higher level of HLA-DR, CD40 and ICOS-L following cytokine stimulation. In addition, HSVECs upregulate the costimulatory ligand CD80 (B7.1) following CD40 ligation, and support allogeneic T cell proliferation, while HUVECs fail to express CD80. Due to differential expression of adhesion molecules, poorly differentiated tumour cell lines also showed more adhesion to HSVECs than to HUVECs. These results indicate that HSVECs have advantages over HUVECs for studying adult vascular endothelial pathology in vitro.

Antibody targeted gene transfer to endothelium.

BACKGROUND: One of the drawbacks of the currently available vectors for gene therapy is the lack of selectivity in gene delivery. We have therefore investigated a strategy to generate immunoliposomes to target non-viral vectors to cell surface receptors on endothelium. MATERIALS AND METHODS: We have developed a novel method of coupling antibodies (Abs) to liposomes complexed to DNA, using mild heat treatment to aggregate the immunoglobulin G (IgG). The interaction of plasmid DNA, liposomes and Abs was measured using a gel retardation assay and a resonant mirror biosensor. The size of the transfection complex was determined by light scattering, and the binding and internalization of the complex to cells was followed using flow cytometry. The transfection ability was tested on cell lines and primary cells in vitro and human corneal or vascular tissues ex vivo. RESULTS: The interaction of antibodies with liposomes is relatively stable (t(1/2) congruent with 45 min). The size of the liposome, Ab and DNA complex was found to be around 500 nm in 4% BSA. The addition of anti-transferrin receptor Abs increased the internalization of the liposome-DNA complex into cells. Abs against both transferrin receptor and E-selectin were shown to augment transfection efficiency of liposomes to cell expressing the appropriate antigens. They are also shown to be efficient in mediating gene delivery to corneal and vascular tissues ex vivo. CONCLUSIONS: We have shown that our novel vector is capable of in vitro and ex vivo gene delivery to cells and human tissues including cornea, artery and vein. In particular, an Ab against E-selectin was effective at selectively delivering genes to activated endothelial cells expressing the adhesion molecule. Such a strategy will have applications for targeting these tissues prior to transplantation or autologous grafting, and, in the longer term, may allow in vivo targeting of gene therapy to inflammatory sites.

ATP-induced activation of expressed RyR3 at low free calcium.

Ca(2+) channel properties of the mink ryanodine receptor type 3 (RyR3), expressed in HEK293 cells, were studied in planar lipid bilayers to which RyR3 rich membrane fragments had been fused. RyR3 channels were not active at resting levels of Ca(2+)(free) but were gated by an additional 1 mM ATP, exhibiting long open times. The second major finding was the absence of channel inactivation at millimolar Ca(2+)(free). Insertion of a myc tag at the N-terminus of RyR3 did not affect the channel properties. As to skeletal muscle, the observed type 3 channel properties appear physiologically meaningful by assisting type 1 channels in calcium release.

Epstein-Barr virus-transformed human B lymphocytes produce natural antibodies to histones.

To study the mechanism(s) responsible for the appearance of Epstein-Barr virus (EBV)-induced anti-histone autoantibodies, peripheral blood B lymphocytes from healthy donors were infected with EBV and the resulting lymphoblastoid cell lines were tested for secretion of antibodies reacting with histones. It was found that EBV-transformed cells produce IgM antibody reactive with histones and that the frequency of EBV-inducible circulating B lymphocytes that produce antibodies to histones is at least 10(-5). Moreover, in cultures of tonsillar lymphoid cells, the enrichment in CD5+ B lymphocytes increases the percentage of EBV-transformed cultures making anti-histone IgM antibodies. EBV may therefore, also in vivo, induce natural anti-histone antibody by polyclonal B-cell activation without any requirement of antigen to trigger antibody response.

Epstein-Barr virus-transformed B lymphocytes produce low molecular mass molecules with autocrine growth factor and competence factor activity.

A human Epstein-Barr virus (EBV)-positive lymphoblastoid B cell line, named BA-D10-4, produces a factor of a molecular mass less than 10 kDa that promotes cell proliferation of both BA-D10-4 cells and other human T or B lymphoid cell lines, either EBV-positive or -negative. The factor synergizes with higher molecular mass autocrine growth factors and makes both BA-D10-4 cells and B cell lines from Burkitt's lymphoma, but not cells from T cell leukemia, more responsive to interleukin-1 and interleukin-6. Therefore, this low molecular mass factor seems to be an autocrine growth factor per se and to have the characteristics of a competence factor.

Density-dependent responsiveness to autocrine growth factors of Epstein-Barr virus transformed human B lymphocytes.

Analysis of the growth requirements of Epstein-Barr virus (EBV)-transformed B lymphocytes shows that interleukin 1 and thioredoxin, a disulfide reducing enzyme, are able to induce a marked increase in DNA synthesis in the early phases of in vitro culture. By contrast, interleukin 6 induces a steady increase in DNA synthesis comparable to that observed with crude conditioned supernatant. Furthermore, EBV-transformed B cells exhibit a density-dependent responsiveness to autocrine growth factors, thus suggesting that growth regulation of EBV-transformed B cells might result from the interplay between different self-stimulating soluble factors and from the competence of the cells to respond to autocrine growth factors.

Antibodies to histones in infectious mononucleosis.

A polyspecific human monoclonal (auto)antibody, isolated from a patient in the acute phase of infectious mononucleosis, was found to react with all subfractions (H1, H2A, H2B, H3 and H4) of histones. This finding prompted us to study the occurrence of antibodies to histones in sera of patients with infectious mononucleosis. It was found that IgM binding to histones was detectable both in control and patient sera; however, sera from patients showed binding values of IgM antibodies to histones significantly higher than those of healthy controls; moreover, both in control and patient groups anti-histone IgM activity was found to correlate with serum IgM concentration. These findings suggest that anti-histone IgM antibodies belong to the class of antibodies defined as "natural antibodies" and that their increase during infectious mononucleosis is due to Epstein-Barr virus-induced polyclonal B cell activation.