Histiocytic sarcoma of the nasal cavity in a horse.

Histiocytic diseases in veterinary medicine have been revised in the last few decades, but these are considered relatively rare in horses. This report describes a 9-year-old female horse, Dutch Warmblood, presented for investigation of severe nasal bleeding. A multinodular bilateral mass of 5 cm, reddish to white in color, that invaded and destroyed the surrounding tissues, was observed during a clinical examination of the nostril The morphological features of the tumor cells were represented by cytologically bizarre, highly phagocytic, multinucleated giant cells. These findings, together with immunohistochemical results allowed a diagnosis of histiocytic sarcoma.

Productive performance and histological features of intestinal mucosa of broiler chickens fed different dietary protein levels.

To evaluate the effect of decreasing dietary protein on growth performance, carcass traits, and intestinal mucosal morphometry, 180 female Hubbard strain broiler chickens were divided into 3 groups and fed 3 isoenergetic diets ad libitum from 14 d of age until slaughter age (49 d). The treatments varied according to 3 protein levels: high-protein diet (HiP, 22.5% CP, DM basis), medium-protein diet (MedP, 20.5% CP), and low-protein diet (LowP, 18.5%). Diets were obtained by replacing wheat middlings with soybean meal and were formulated to meet or exceed broiler amino acid requirements of the NRC. Morphometric indices of duodenum, jejunum, and ileum were measured at the end of the feeding period and included villus height, crypt depth, villus-to-crypt ratio, and apparent villus surface area. The dietary protein level had a significant effect on final BW of birds, whereas ADG, ADFI, and feed efficiency remained unaffected by dietary treatment. The muscle (breast and drumstick) yields were significantly higher in birds fed the HiP diet compared with those of the MedP and LowP diets. Meat quality traits were not affected by the protein level. The villus surface area of all intestinal segments did not change among groups. Instead, reducing the dietary protein level to 20.5% resulted in a higher villus height and villus height to crypt depth ratio in the duodenum and ileum. On the basis of our findings, even if the high-protein diet promoted meat yield, a medium-protein diet could positively support broiler growth performance, as confirmed by favorable morphometric features of the intestine.

Lesions associated with mineral deposition in the lymph nodes and lungs of cattle: a case-control study of environmental health hazard.

This report focuses on the state of health of the cattle raised in the district of Taranto - city of Italy rated as environmentally at risk. Representative samples of lungs, bronchial and mediastinal lymph nodes of cattle from district of Taranto's slaughterhouses were collected. After a macroscopic examination, samples with marked lesions were processed for light microscopy. Samples were also observed with polarized light microscopy, scanning electron microscopy and with microanalysis. The macroscopic examination revealed that 60 out of 183 samples showed marked lesions. Lung alterations were characterized by thickening of the alveolar septa and by the latter's modifying action on the alveolar spaces, foci of fibrosis and bronchopulmonary inflammation. For 51 out of the 60 samples observed, the histological examination confirmed the presence of pneumoconiosis and lymph nodal anthracosis. Energy-dispersive X-ray microanalysis of lung samples identified a wide range of elements including silicon, aluminium, titanium, iron, carbon and small amount of the other metals. In the lymph-nodes the same kind of metals with a different levels of distribution was observed. Our survey on cattle farmed in areas at high risk of pollution may be helpful to the estimation of the exposure risk for man to environmental contaminants and to the evaluation of the occurrence of the pathological manifestations as well.

Multiple glomus tumors of the urinary bladder in a cow associated with bovine papillomavirus type 2 (BPV-2) infection.

We report a case of multiple glomus tumors associated with bovine papillomavirus type 2 (BPV-2) infection in the urinary bladder of a 13-year-old cow suffering from severe chronic enzootic hematuria. Macroscopically, multiple submucosal reddish nodules were seen swelling the vesical mucosa. Histologically, neoplastic proliferation was characterized by the presence of numerous blood vessels. These were lined by normal endothelial cells surrounded by round epithelioid cells with central nuclei, prominent nucleoli, acidophilic cytoplasm, and well-defined cytoplasmic borders. Tumor cells were distributed around open vascular lumina and in perivascular spaces. They were immunohistochemically positive for actin and vimentin and negative for cytokeratins, desmin, and factor VIII-related antigen. On the basis of these findings, this tumor was diagnosed as glomus tumor, a neoplasm not previously reported in cattle and exceedingly rare in animals. BPV-2 DNA was amplified from the formalin-fixed, paraffin-processed tissue specimens obtained by laser capture microdissection. This report widens the spectrum of mesenchymal tumors of the bovine urinary bladder. Finally, the microscopic pattern of tumor described here shares striking morphologic and immunohistochemical similarities with the angiomatous form of glomus tumor known to occur in man.

Source of cell injected is a critical factors for short and long engraftment in xeno-transplantation.

This study aims to investigate engraftment of human cord blood and foetal bone marrow stem cells after in utero transplantation via the intracoelomic route in the sheep. Here, we performed transplantation in 14 single and 1 twin sheep foetuses at 40-47 days of development, using a novel schedule for injection. (i) Single injection of CD34(+) human cord blood stem cells via the coelomic route (from 10 to 50 x 10(4)) in seven single foetuses. (ii) Single injection of CD34(+) foetal bone marrow stem cells via the intracoelomic route with further numbers of cells (20 x 10(5) and 8 x 10(5), respectively) in three single and in one twin foetuses. (iii) Double fractioned injection (20-30 x 10(6)) via the coelomic route and 20 x 10(6) postnatally, intravenously, shortly after birth of CD3-depleted cord blood stem cells in four single foetuses. In the first group, three single foetuses showed human/sheep chimaerism at 1, 8 and 14 months after birth. In the second group, the twin foetuses showed human/sheep chimaerism at 1 month after birth. In the third group, only two out of four single foetuses that underwent transplantation showed chimaerism at 1 month. While foetal bone marrow stem cells showed good short-term engraftment (1 month after birth), cord blood stem cells were able to persist longer in the ovine recipients (at 1, 8 and 14 months after birth).

Expression of CD133-1 and CD133-2 in ovarian cancer.

Cancer stem cells have been isolated from several solid tumors including prostate, colon, liver, breast, and ovarian cancer. Stem cells isolated from nervous system and prostate express CD133 antigen, which is widely used to isolate hematopoietic stem and progenitor cells. The aims of this study were to investigate the expression of the CD133-1 and CD133-2 epitopes in primary ovarian tumors and to biologically characterize CD133(+) ovarian cancer cells, also according to clinicopathologic parameters. Tissue specimens were obtained at primary surgery from 41 ovarian carcinomas; eight normal ovaries and five benign ovarian tumors were also collected. Flow cytometry with monoclonal antibodies against CD133-1 and CD133-2 epitopes was employed. FACS (fluorescence activated cell sorting) analysis enabled the selection of CD133(+) cells, whose epithelial origin was confirmed by immunofluorescence analysis with monoclonal anti-cytokeratin 7. CD133(+) cells gave rise to a 4.7 +/- 0.9-fold larger number of colonies than that documented in CD133(-) population (P < 0.001). Moreover, CD133(+) cells showed an enhanced proliferative potential compared to CD133(-) cells. The percentages of CD133-1- and CD133-2-expressing cells were significantly lower in normal ovaries/benign tumors with respect to those in ovarian carcinoma. Both the percentages of CD133-1- and CD133-2-expressing cells were significantly lower in omental metastases than in primary ovarian cancer (P = 0.009 and 0.007 for CD133-1- and CD133-2-expressing cells, respectively). There seems not to be any difference in the distribution of the percentage of CD133-1- and CD133-2-expressing cells according to clinicopathologic parameters and response to primary chemotherapy. CD133-1 and CD133-2 may be useful in order to select and enrich the population of CD133(+) ovarian tumor cells, which are characterized by a higher clonogenic efficiency and proliferative potential.

In vitro elicitation of intestinal immune responses in teleost fish: evidence for a type IV hypersensitivity reaction in rainbow trout.

In fish the gut immune system has been the subject of few investigations until now. Here, we provide novel morphological and immunological data on the gut isolated from rainbow trout Salmo gairdneri. The pyloric (P) and terminal (T) segments of trout gut, when morphologically examined, evidenced lymphocytes and macrophages (MØ) loosely dispersed in the intestinal mucosa and in the lamina propria in the absence of typical Peyer's patches-like structures. Furthermore, incubation of P and T sections with Candida albicans (Ca) and functional analysis of supernatants generated some interesting results. In fact, active supernatants, when compared with controls, exhibited cytokine-like activities attributable to the presence of interferon (IFN)-gamma and migration inhibiting factor (MIF), respectively. In particular, IFN-gamma-like activity gave rise to an enhancement of Ca phagocytosis by MØ, whereas MIF inhibited MØ migration in agarose. Taken together, these in vitro data suggest that the gut-associated lymphoreticular tissue in fish possesses the appropriate armamentarium to mount a type IV hypersensitivity response when challenged by microbial antigens.

First observation of an Hashimoto thyroiditis-like disease in horses from Eastern Europe: histopathological and immunological findings.

Among 622 slaughtered horses from eastern Europe, 156 thyroid glands were selected on the basis of macroscopic alterations (e.g., determination of volume and weight). In the 80% of these thyroids, microscopic alterations consistent with a diagnosis of Hashimoto thyroiditis-like disease were found. In particular, a subverted architecture of the thyroid gland with colloid rarefaction, lymphocytic infiltration, and fibrosis was noted. The confirmation of the histopathological diagnosis of an equine Hashimoto thyroiditis-like disease was provided by the increased serum concentration of thyroglobulin, of antithyroglobulin, and of antithyroid peroxidase autoantibodies. Despite evidence consistent with an Hashimoto thyroiditis-like disease in eastern European horses, the etiopathogenesis of this autoimmune disorder deserves further investigation. In this respect, in some horses histopathological alterations in the pituitary gland may suggest an as-yet-unidentified disorder within the hypothalamus-pituitary adrenal axis associated with Hashimoto thyroiditis.

Modifications of serum and cellular parameters in trotters after a race. Macrophage migration inhibitory activity reduction and serum beta-glucan elevation.

Trotters are exposed to a chronic prolonged stress, such as daily training and frequent races during their active lifespans. There is evidence that trotters undergo very often lethal lung infections after a race, and therefore, is likely that modifications of certain physiologic cellular parameters could account for the increased susceptibility to microbial diseases. Here, we demonstrate that in 7 trotters after a race either serum values (e.g., glycaemia, triglycerides, transaminases, gamma-glutamyltransferase, cholinesterase, amylase, alkaline phosphatase, total proteins, serum albumin, sodium, blood urea nitrogen, lactic dehydrogenase, creatine phosphokinase, and creatinine) or hematological parameters (red blood cell count, hemoglobin, lymphocyte and monocyte count) were increased. At the same time, in the same animals after a race, macrophage migration inhibitory factor activity was depressed, thus indicating an impaired T-lymphocyte response. Finally, increased levels of circulating beta-glucans in some horses, after a race, may suggest a reduced clearance of fungal cell wall components. Taken together, these findings indicate a condition of multiple organ dysfunction, such as the liver, the kidney, the pancreas, and skeletal muscles, as well as a reduced cell-mediated immune response in trotters, after a race.

Stem cell-based treatments for gynecological solid tumors.

BACKGROUND AND OBJECTIVE: We have recently assisted to an increasing scientific interest and a new research effort in the field of stem cell-based therapy. Since the late 1980s hematopoietic stem cells (HSC) have been used to set up therapeutic strategies for the treatment of solid tumors such as gynecological cancers. In this context, different approaches have been suggested and clinically investigated. STATE OF THE ART: In the autologous setting we can describe the well-known use of HSC as hematologic support to high-dose chemotherapy regimens, and the use of HSC as a source of dendritic cells for cancer vaccination protocols. In our institution a long-term experience has been developed in high-dose chemotherapy with autologous HSC transplantation as first-line treatment of advanced ovarian cancer, and in the use of cytokines both for HSC collection and for post-transplantation hematopoietic recovery and immune reconstitution. An alternative approach consists of allogenic HSC transplantation following either myeloablative/standard or non-myeloablative/reduced conditioning regimens, which have been proposed as new adoptive immunotherapeutic treatments for different non-hematologic malignancies. PERSPECTIVES: Future strategies in the use of HSC in oncology comprise the possibility of HSC ex-vivo expansion, the use of umbilical cord blood HSC, and the development of HSC-based gene-therapy programs. Further investigations are expected in the new field of cancer stem cells.

[High-dose chemotherapy for metastatic breast cancer]. / La chemioterapia ad alte dosi nel carcinoma della mammella metastatico.

In ninety's breast cancer was first in Europe for the use of high-dose chemotherapy with autologous hematopoietic stem cell transplantation in solid tumors in adults. Some phase II trials of high-dose chemotherapy showed high response rates and prolonged progression free survival in selected metastastic breast cancer patients. Few large, powerful randomized phase III studies comparing this approach with conventional chemotherapy have been completed: some studies showed a better progression free survival in favor of high dose chemotherapy, but no statistically significant difference in overall survival was observed. Many variables inside high dose chemotherapy program need to be considered. The identification of subsets of breast cancer patients who can benefit from high-dose chemotherapy is essential: high-dose chemotherapy should be included in a global treatment strategy, evaluating the integration with innovative treatment modalities, with the aim of eradicating the minimal residual disease in breast cancer patients achieving complete response after high dose chemotherapy.

Stem cells in gynecology and obstetrics.

Over the past 10 years, we have become involved in a new research effort and an increasing scientific interest in the field of stem cell-based therapy. We are therefore able to describe different areas in which stem cell research can be applied and developed in gynecology and obstetrics. I) Hematopoietic stem cells have been used to set up therapeutic strategies for the treatment of gynecological solid tumors such as ovarian cancer. In this context different autologous or allogeneic transplantation approaches have been proposed and clinically investigated. II) Umbilical cord blood, which was often considered a waste material of the delivery, actually represents a precious source of stem cells that can be used for cell-based treatments of malignancies and inherited diseases. III) A feto-maternal cell traffic has recently been demonstrated through the placental barrier during pregnancy. This cellular exchange also includes stem cells from the fetus, which can generate microchimerisms in the mother and contribute to tissue repair mechanisms in different maternal organs. IV) Stem cells can be used for prenatal transplantation to treat different severe congenital diseases of the fetus. Nevertheless, several problems need to be solved to achieve an efficient in utero stem cell transplantation. Recent reports have pointed out the importance of timing in prenatal stem cell transplantation procedures and have shown the advantage of an early stem cell injection. An ultrasound-guided intracelomic approach could allow this possibility.

Cyclooxygenase-1 and -2 expression in urothelial carcinomas of the urinary bladder in cows.

Bovine urinary bladder tumors occur frequently in animals suffering from chronic enzootic hematuria because of prolonged ingestion of bracken fern (Pteridium spp.). Cyclooxygenase (COX) genes (COX-1 and COX-2) are known to be involved in the carcinogenesis of human and some animal urothelial tumors. The aim of the present study was to investigate COX-1 and COX-2 expression by immunohistochemical methods in 20 bovine urothelial carcinomas collected at public slaughterhouses from cows that had been suffering from chronic enzootic hematuria. COX-1 immunostaining was identified intracytoplasmically in normal urothelium and in 15 of 20 neoplastic specimens. COX-1 immunosignal in the tumor cells was either absent or weak. COX-2 was also expressed intracytoplasmically in 17 of 20 urothelial carcinomas. Moderate to intense COX-2 labeling was detected in both noninvasive and invasive urothelial carcinomas. Coexpression of both enzyme isoforms was also revealed by confocal laser scanning microscopic investigations. This study indicates that COX-2 is overexpressed in naturally occurring urothelial carcinomas of cows.

Administration of low-dose interleukin-2 plus G-CSF/EPO early after autologous PBSC transplantation: effects on immune recovery and NK activity in a prospective study in women with breast and ovarian cancer.

This study evaluated the effects of low-dose IL-2 plus G-CSF/EPO on post-PBSC transplantation (PBSCT) immune-hematopoietic reconstitution and NK activity in patients with breast (BrCa) and ovarian cancer (OvCa). To this end, two consecutive series of patients were prospectively assigned to distinct post-PBSCT cytokine regimens (from day +1 to day +12) which consisted of G-CSF (5 microg/kg/day) plus EPO (150 IU/kg/every other day) in 17 patients (13 BrCa and 4 OvCa) or G-CSF/EPO plus IL-2 (2 x 10(5) IU/m(2)/day) in 15 patients (10 BrCa and 5 OvCa). Hematopoietic recovery and post-transplantation clinical courses were comparable in G-CSF/EPO- and in G-CSF/EPO plus IL-2-treated patients, without significant side-effects attributable to IL-2 administration. In the early and late post-transplant period a significantly higher PMN count was observed in G-CSF/EPO plus IL-2-treated patients (P = 0.034 and P = 0.040 on day +20 and +100, respectively). No significant differences were found between the two groups of patients in the kinetics of most lymphocyte subsets except naive CD45RA(+) T cells which had a delayed recovery in G-CSF/EPO plus IL-2 patients (P = 0.021 on day +100). No significant difference was observed between NK activity in the two different groups, albeit a significantly higher NK count was observed in G-CSF/EPO plus IL-2 series on day +20 (P = 0.020). These results demonstrate that low-dose IL-2 can be safely administered in combination with G-CSF/EPO early after PBSCT and that it exerts favorable effects on post-PBSCT myeloid reconstitution, but not on immune recovery.

The role of hematopoietic stem cells in the treatment of ovarian cancer.

In recent years hematopoietic stem cells (HSC) have been the object of new research efforts and scientific advances. Therapeutic strategies have been set up using HSC for the treatment of solid tumors such as ovarian cancer. In this context different approaches have been proposed and clinically investigated. The "autologous" approach refers to the use of HSC as hematologic support to high-dose chemotherapy regimens, and to the use of HSC as an abundant source of dendritic cells for cancer vaccination protocols. Our institution has developed a long-term experience in high-dose chemotherapy with autologous HSC transplantation as first-line treatment of advanced ovarian cancer, and in the use of cytokines both for the HSC collection and for the post-transplantation hematopoietic recovery. Moreover, the "allogeneic" approach with HSC consists of the allogeneic transplantation with both myeloablative/standard or nonmyeloablative/reduced conditioning regimens, which has been proposed as a new adoptive immunotherapeutic treatment for different nonhematologic malignancies. Perspectives in the use of HSC in oncology comprise the possibility of an HSC ex vivo expansion, the use of umbilical cord blood HSC, and the development of future HSC-based gene-therapy programs.

Docetaxel and epirubicin plus G-CSF as mobilizing treatment to support high-dose chemotherapy in breast cancer.

BACKGROUND: In order to combine an active regimen with a simultaneous efficient mobilization of peripheral blood precursor cells (PBPC), we explored the combination of Docetaxel 75 mg/m2 and Epirubicin 120 mg/m2 with G-CSF 5 mcg/Kg/day s.c. to mobilize PBPC in breast cancer patients to support high-dose chemotherapy (HDC). PATIENTS AND METHODS: Forty patients were enrolled: 27 high risk and 13 metastatic. The entire procedure, including chemotherapy and PBPC collection, was on an outpatient basis. RESULTS: The median day of starting apheresis was day +10 (range 10-12) and the average value of circulating CD34+ cells at peak was 175/microliter (range 33-403). The median yield of CD34+ cells per apheresis was 8.76 x 10(6)/Kg (range 1.83-27.87). None of the patients developed side effects which required hospitalization. All patients enrolled successively received HDC as consolidation treatment. High risk patients received one and metastatic patients two HDC with PBPC reinfusion. All patients obtained a complete engraftment. No significant differences between high-risk and metastatic patients were observed. CONCLUSIONS: Our study suggests that the combination of Docetaxel, Epirubicin, and G-CSF is feasible, safe and efficient outpatient mobilizing treatment for patients with breast cancer receiving HDC.

Peripheral blood progenitor cell collection after epirubicin, paclitaxel, and cisplatin combination chemotherapy using EPO-based cytokine regimens: a randomized comparison of G-CSF and sequential GM-/G-CSF.

BACKGROUND: The peripheral blood progenitor cell (PBPC) mobilization capacity of EPO in association with either G-CSF or sequential GM-CSF/G-CSF was compared in a randomized fashion after epirubicin, paclitaxel, and cisplatin (ETP) chemotherapy. STUDY DESIGN AND METHODS: Forty patients with stage IIIB, IIIC, or IV ovarian carcinoma were enrolled in this randomized comparison of mobilizing capacity and myelopoietic effects of G-CSF + EPO and GM-/G-CSF + EPO following the first ETP chemotherapy treatment. After ETP chemotherapy (Day 1), 20 patients received G-CSF 5 microg per kg per day from Day 2 to Day 13 and 20 patients received GM-CSF 5 microg per kg per day from Day 2 to Day 6 followed by G-CSF 5 microg per kg per day from Day 7 to Day 13. EPO (150 IU per kg) was given every other day from Day 2 to Day 13 to all patients in both arms of the study. Apheresis (two blood volumes) was performed during hematologic recovery. RESULTS: The magnitude of CD34+ cell mobilization and the abrogation of patients' myelosuppression were comparable in both study arms; however, GM-/G-CSF + EPO patients had significantly higher CD34+ yields because of a higher CD34+ cell collection efficiency (57.5% for GM-/G-CSF + EPO and 46.3% for G-CSF + EPO patients; p = 0.0009). Identical doses of PBPCs mobilized by GM-/G-CSF + EPO and G-CSF + EPO drove comparable hematopoietic recovery after reinfusion in patients treated with identical high-dose chemotherapy. CONCLUSION: The sequential administration of GM-CSF and G-CSF in combination with EPO is feasible and improves the PBPC collection efficiency after platinum-based intensive polychemotherapy, associating high PBPC mobilization to high collection efficiency during apheresis.