Modulation of the effects of lung immune response on bone marrow by oral antigen exposure.

Allergic airway inflammation is attenuated by oral tolerization (oral exposure to allergen, followed by conventional sensitization and challenge with homologous antigen), which decreases airway allergen challenge-induced eosinophilic infiltration of the lungs and bone marrow eosinophilia. We examined its effects on bone marrow eosinophil and neutrophil production. Mice of wild type (BP-2, BALB/c, and C57BL/6) and mutant strains (lacking iNOS or CD95L) were given ovalbumin (OVA) or water (vehicle) orally and subsequently sensitized and challenged with OVA (OVA/OVA/OVA and H2O/OVA/OVA groups, resp.). Anti-OVA IgG and IgE, bone marrow eosinophil and neutrophil numbers, and eosinophil and neutrophil production ex vivo were evaluated. T lymphocytes from OVA/OVA/OVA or control H2O/OVA/OVA donors were transferred into naïve syngeneic recipients, which were subsequently sensitized/challenged with OVA. Alternatively, T lymphocytes were cocultured with bone marrow eosinophil precursors from histocompatible sensitized/challenged mice. OVA/OVA/OVA mice of the BP-2 and BALB/c strains showed, relative to H2O/OVA/OVA controls, significantly decreased bone marrow eosinophil counts and ex vivo eosinopoiesis/neutropoiesis. Full effectiveness in vivo required sequential oral/subcutaneous/intranasal exposures to the same allergen. Transfer of splenic T lymphocytes from OVA/OVA/OVA donors to naive recipients prevented bone marrow eosinophilia and eosinopoiesis in response to recipient sensitization/challenge and supressed eosinopoiesis upon coculture with syngeneic bone marrow precursors from sensitized/challenged donors.

Inhibition of long-term potentiation by CuZn superoxide dismutase injection in rat dentate gyrus: involvement of muscarinic M1 receptor.

Long-term potentiation (LTP) and long-term depression represent important processes that modulate synaptic transmission that carries out a key role in neural mechanisms of memory. Many studies give strong evidences on a role of the reactive oxygen species in the induction of LTP in CA1 region of hippocampal slices that was inhibited by adding the scavenger enzyme superoxide dismutase (SOD1). Previous data showed that SOD1 is secreted by many cellular lines, including neuroblastoma SK-N-BE cells through microvesicles by an ATP-dependent mechanism; moreover, it has been shown that SOD1 interacts with human neuroblastoma cell membranes increasing intracellular calcium levels via a phospholipase C-protein kinase C pathway activation. The aim of this study was to investigate the effect of intracerebral injection of SOD1 or the inactive form of enzyme (ApoSOD) on the modulation of synaptic transmission in dentate gyrus of the hippocampus in urethane anesthetized rats. The results of the present research showed that intracerebral injection of SOD1 and ApoSOD in the dentate gyrus of the rat hippocampal formation inhibits LTP induced by high-frequency stimulation of the perforant path. This result cannot be only explained by the dismutation of oxygen radical induced by SOD1 since also ApoSOD, that lacks the enzymatic activity, carries out the same inhibitory effect on LTP induction.

Intracerebroventricular injection of oxidant and antioxidant molecules affects long-term potentiation in urethane anaesthetized rats.

Production of superoxide anions in the incubation medium of hippocampal slices can induce long-term potentiation (LTP). Other reactive oxygen species (ROS) such as hydrogen peroxide are able to modulate LTP and are likely to be involved in aging mechanisms. The present study explored whether intracerebro-ventricular (ICV) injection of oxidant or antioxidant molecules could affect LTP in vivo. With this aim in mind, field excitatory post-synaptic potentials (fEPSPs) elicited by stimulation of the perforant pathway were recorded in the dentate gyrus of the hippocampal formation in urethane-anesthetized rats. N-acetyl-L-cysteine, hydrogen peroxide (H2O2) or hypoxanthine/xanthine-oxidase solution (a superoxide producing system) were administrated by ICV injection. The control was represented by a group injected with saline ICV. Ten minutes after the injection, LTP was induced in the granule cells of the dentate gyrus by high frequency stimulation of the perforant pathway. Neither the H(2)O(2) injection or the N-acetyl-L-cysteine injection caused any variation in the fEPSP at the 10-min post-injection time point, whereas the superoxide generating system caused a significant increase in the fEPSP. Moreover, at 60 min after tetanic stimulation, all treatments attenuated LTP compared with the control group. These results show that ICV administration of oxidant or antioxidant molecules can modulate LTP in vivo in the dentate gyrus. Particularly, a superoxide producing system can induce potentiation of the synaptic response. Interestingly, ICV injection of oxidants or antioxidants prevented a full expression of LTP compared to the saline injection.

Immunohistochemical distribution of proteins belonging to the receptor-mediated and the mitochondrial apoptotic pathways in human placenta during gestation.

The balance between cell death and cell proliferation and its regulation are essential features of many physiological processes and are particularly important in fetal morphogenesis and adult tissue homeostasis. Apoptosis is a type of cell suicide that is activated in two main ways: through a receptor-mediated pathway or through a mitochondrial pathway. We have investigated the immunohistochemical distribution of proteins belonging to these two pathways in human placenta during gestation by comparing their expression levels between the first and third trimester of gestation. In the first trimester, the receptor-mediated pathway prevails over the mitochondrial pathway with a moderate/intense expression of its three components, viz., Fas ligand (FasL), Fas, and caspase-8, and weak positivity of anti-apoptotic FLIP, these proteins being mainly localized in the cytotrophoblast compartment. In the third trimester of gestation, there is an increased expression of mitochondrial pathway proteins, viz., Apaf-1 and caspase-9. We have also investigated the expression level of caspase-3, the primary effector caspase of both pathways, and have observed that it is moderately expressed during gestation, being mainly localized in the cytotrophoblast during the first trimester and in both placental compartments during the third trimester of gestation. Thus, both pathways actively function in human placenta to execute cell death. By means of immunoelectron microscopy, we have further shown that, in human placenta, the two proteins of the mitochondrial pathway together with caspase-3 are localized both in the cytoplasm and in the nucleus. In particular, Apaf-1 and caspase-9 are distributed near to the nuclear envelope suggesting an important role for these two proteins in disrupting the nuclear-cytoplasmic barrier.

Detachable snare versus epinephrine injection in the prevention of postpolypectomy bleeding: a randomized and controlled study.

BACKGROUND AND STUDY AIMS: Several endoscopic techniques have been developed to prevent bleeding after the removal of large pedunculated polyps. PATIENTS AND METHODS: From January 1995 to December 2002, 488 consecutive patients with pedunculated colorectal polyps, the heads of which were larger than 10 mm in diameter, were randomly assigned to three groups. In group A (163 patients), detachable snares were placed at the base of the stalk and standard snares were then used for polypectomy. In group B (161 patients), the polyp stalk was injected with a 0.01 % epinephrine solution before conventional snare polypectomy. Group C (a control group including 164 patients) underwent conventional snare polypectomy without preventive measures. Early (< 24 h) and late (> 24 h - 30 days) bleeding complications were assessed. Each group was divided into two subgroups relative to the polyp size (polyps 1.0 - 1.9 cm and polyps > or = 2 cm). RESULTS: Overall bleeding complications occurred after 4.3 % of the polypectomies. Bleeding was successfully controlled in all patients, and no blood transfusions were required. There were three cases of bleeding in group A (1.8 %), five in group B (3.1 %), and 13 in group C (7.9 %). Early bleeding was more frequent than late bleeding (15 vs. six patients). In polyps > or = 2 cm (207 patients), postpolypectomy bleeding occurred in 14 patients (6.7 %): two (2.7 %) in the detachable snare group, two (2.9 %) in the epinephrine injection group, and 10 (15.1 %) in the control group. CONCLUSIONS: These results show that polypectomy of large pedunculated polyps is associated with a higher incidence of bleeding. Particularly in polyps larger than 2 cm, preventive measures can significantly reduce bleeding complications after polypectomy. This can be achieved with similar efficacy either by placing Endoloops or by injecting epinephrine.

Immunohistochemical expression of cyclin T1 in a case of AIDS-related cachexia.

The expression of cyclin T1 in an autoptic case of AIDS-related cachexia was investigated by immunohistochemistry. When contrasted with normal human tissues, a very similar pattern of expression was found. However, a peculiar distribution of cyclin T1 was noticed in the brown fat and in lymph nodes affected by AIDS-associated lymphadenopathy.

Expression of the retinoblastoma-related p107 and Rb2/p130 genes in human placenta: an immunohistochemical study.

It has been proposed that tumor suppressor genes may have a role in the mechanisms of proliferation and differentiation during human placental development. The Retinoblastoma gene family is a well known family of tumor suppressor genes. Many studies have pointed out a role of this family not only in cell cycle progression, but also during development and differentiation. On the light of these observations we have investigated the immunohistochemical expression pattern of the Retinoblastoma family members, p107 and Rb2/p130 in human placenta samples in first trimester and full-term placental sections. p107 and pRb2/p130 showed the most abundant expression levels during the first trimester of gestation and progressively declined to being barely detectable in the placenta by late gestation. These results indicate that the expression of the above genes is modulated during placental development and suggest a mechanism for controlling trophoblast proliferation.

Alteration of the Bcl-2:Bax ratio in the placenta as pregnancy proceeds.

The placenta is the primary site of nutrient and gas exchange between mother and foetus. During human placental development, proliferation, differentiation and apoptosis occur at different stages. In order to clarify some of the molecular mechanisms underlying these events, we investigated the pattern of expression of two members of the Bcl-2 family in human placenta samples and compared them to the level of apoptosis detected by the TUNEL method. In particular, we evaluated the expression of Bcl-2 and Bax and their ratio during the first and third trimester. We found that Bcl-2 was generally expressed at low levels during the entire gestational period. On the other hand, Bax was low during the first trimester but increased towards the end of gestation. In accordance with the change of ratio of these two molecules, the increase of apoptotic cells was observable in the third trimester. These data indicate that Bcl-2 and Bax are spatio-temporally regulated during placental development and that the different expression of the above mentioned genes is at least in part responsible for the delicate balance between cell proliferation and programmed cell death in the human placenta during pregnancy.

Macrophages and antioxidant status in the NOD mouse pancreas.

This study showed that citiolone (CIT), a free radical scavenger, significantly increased superoxide dismutase (P < 0.001 vs. untreated NOD, NMMA-treated, and silica-treated animals), catalase (P < 0.01 vs. untreated NOD), and glutathione peroxidase (P < 0.001 vs. untreated NOD and C57BL6/J) values. Silica treatment was capable of counteracting the plasma antioxidant capacity (TRAP) decrease observed in untreated NOD mice, although it did not block the blood glucose rise and insulitis progression in type 1 diabetes significantly. Conversely, early silica administration was able to deplete macrophages (as demonstrated by immunocytochemistry) and to block the rise in blood glucose levels and insulitis progression significantly. Silica-treated animals in this study showed the highest TRAP levels, demonstrating that depletion of macrophages also was able to improve the antioxidant status. This study suggested that macrophages are essential for type 1 diabetes development and showed that they also are involved when the antioxidant status is affected. The reported findings are significant in view of previous studies indicating that oxygen and/or nitrogen free radicals contribute to the islet beta-cell destruction in type 1 diabetes animal models.

Prognostic role of cyclin D1 in non small cell lung cancer: an immunohistochemical analysis.

Lung cancer is a worldwide problem and in many countires it is the most lethal malignancy. Because relapse is frequent after resection of non small cell lung cancer, an urgent need exists to define prognostic factors which could help in choosing the best therapeutic approach. We performed immunohistochemistry on 60 formalin-fixed paraffin-embedded non small cell lung cancer specimens in order to evaluate the frequency of cyclin D1 overexpression, and to relate it to the degree of malignancy of these tumors and to the overall survival time of the patients. All specimens were positive for cyclin D1 immunostaining. We found cyclin D1 overexpression in 30 (50%) of our specimens, with no significant difference among the different histological types. Cyclin D1 overexpression correlates in a statistical manner with short-term patient survival. Mantel-Cox analysis of these data generated a significant P value = 0.003. The mean survival time and the five-year survival rate also differed statistically. We did not find any statistically significant correlation between cyclin D1 overexpression and histological grading, tumor stage or TNM status. We concluded that cyclin D1 overexpression in 30 patients is a frequent event in non small cell lung cancer pathogenesis and may have prognostic relevance.

[The diagnostic significance of the small bowel enema in lesions of the small intestine requiring surgery]. / Importanza diagnostica del clisma del tenue nelle lesioni di interesse chirurgico del piccolo intestino.

Given its anatomical location the small intestine is the section of the digestive tract most inaccessible to objective clinical and instrumental examination. For this reason, traditional investigation methods provide inadequate data. In the barium meal X-ray, images are superimposed and the barium excessively fragmented due to the tortuosity and length of the intestinal loop. Access problems limit the value of endoscopy. The indications to angiography are fairly restrictive. Ultrasound and CT scans provide no conclusive evidence in this area. The so-called Small Bowel Enema technique based on the direct infusion of an opaque contrast medium after duodenal intubation remains the only way to obtain images with a satisfactory diagnostic value and should be the method of choice in examinations of the small intestine. Introduced in the twenties, the value of the technique was long questioned, mainly because it was difficult to perform correctly and it is only quite recently that several decisive technological innovations have made it highly reliable. The present paper describes its use on a series of patients with suspected pathologies of the small intestine requiring surgical treatment. The aim is to specify the correct indications to this technique as well as its limitations with a view to promoting its appropriate use in routine practice. MATERIALS AND METHODS. The study covers 42 small bowel enemas performed on 29 male and 13 female patients aged 12-85 years. Indications to the enema included chronic abdominal pain in 28.5%, chronic diarrhea in 21.4%, pain+diarrhea in 14%, subocclusion in 16.5%, faecal blood in 11.9%, abdominal pain+diarrhea+intestinal haemorrhage in 4.7%, anaemia in 2.4%. We adopted the Sellink technique involving intubation with a French 14 Bilbao-Dotter probe and the infusion of 200-350 cc barium sulphate in a 90%-70% P/V concentration at a speed of 100 ml/min, followed by the infusion of 1000-1500 cc 0.5-1% methylcellulose in a 50-50 mixture with water for double contrast purposes. Mean duration of procedure 35-40 mins. RESULTS. We found lesions of the small intestine in 18 cases or 45% (true positives) and no alterations in 18 or 42.8% (true negatives). In 3 cases (7%) the examination was inconclusive. One false positive and one false negative complete the series, making a diagnostic accuracy of 89.1%. In 23.8% of the total series the diagnosis of Crohn's disease was confirmed; cancerous lesions were found in 11.9% and adhesions following earlier surgery in 9.7%. Surgery confirmed this diagnosis in 10 cases (5 tumours, 3 stenoses caused by adhesion fragments, 2 Crohn's disease) and disproved it in one case. Biopsy was used, for objective confirmation in 5 cases (2 Crohn's disease, 3 negatives). In the other cases reliance was placed on clinical course and the patients' response to treatment. DISCUSSION AND CONCLUSIONS. Examination of our results reveals that in most clinical cases the small bowel enema was used to confirm suspected Crohn's disease (31 cases out of 42) and with a very high degree of diagnostic accuracy (96.7%). In line with data in the literature we decided to base the indication to this examination on certain primary symptoms such as chronic abdominal pain, diarrhea, blood in the faeces, whether in isolation or in combination. On this rather general basis the diagnosis of Crohn's disease concerned only 23.8% of the cases studied, an incidence that rose dramatically in the few cases in which the indications were made more specific. We therefore conclude that the adoption of more restrictive criteria will improve the cost-benefit ratio for this test. Another major pathology in which small bowel enema possesses a high diagnostic value is partial obstruction of the small intestine.(ABSTRACT TRUNCATED AT 400 WORDS)

Heat production and motor deficit in rats lesioned in globus pallidus, entopeduncular nucleus and lateral hypothalamus.

Oxygen consumption, colonic and interscapular brown adipose tissue temperature were evaluated in four groups of male and four groups of female rats both before and after lesions in different brain regions, and following beta-blocker propranolol administration. Recovery of body weight with varying difficulties in reaching food was also recorded in the injured animals. Groups consisted of rats with bilateral lesions in the entopeduncular nucleus (group EN), rats with bilateral lesions in the globus pallidus (group GP), rats with lateral hypothalamic lesions in the left side and entopeduncular lesion in the right side (group EN-LH), and rats with bilateral lesions in the lateral hypothalamus (group LH). Colonic and brown adipose tissue temperature and oxygen consumption were significantly increased after lesions in rats of groups EN, LH and EN-LH, but not in animals of the GP groups. Similarly, propranolol administration blocked the rise in heat production only in EN, LH and EN-LH animals. No differences were found between sexes. The survival rate was the same in all groups. GP rats recovered body weight earlier than animals injured in the other regions. The difficulty in reaching food was an important factor only in rats damaged in the EN. The results suggest that lateral hypothalamus and entopeduncular nucleus share a common regulatory function of the energy metabolism, while EN lesions induce a motor deficit in addition.

Phase II double-blind randomized study of lonidamine and radiotherapy in epidermoid carcinoma of the lung.

Patients with non metastatic squamous cell lung cancer were treated with radiotherapy (RT) plus lonidamine (LND) or placebo (PLAC), according to a randomized double-blind study design. Treatment with lonidamine 150 mg t.i.d. (27 patients) or placebo (23 patients) started 3 days before RT, lasted up to 7 months. Partial responses were observed in 14 and 6 patients respectively in the LND + RT and PLAC + RT groups. Statistical analysis of the survival curves showed no significant difference between the LND + RT (median 311 days) and PLAC + RT (median 193 days) groups. Stage III patients survived significantly longer (p less than 0.05) when treated with LND + RT (median 318 days) than with PLAC + RT (median 163 days). No synergistic toxic effects between radiation and LND were noted. To confirm these data a new and larger multicentric study is now in progress.

Cortical control of thermogenesis induced by lateral hypothalamic lesion and overeating.

Increased O2 consumption was found in rats after bilateral lesions of the lateral hypothalamus (LH) or during voluntary overeating. This phenomenon appears to be mediated by the sympathetic nervous system (SNS) in both conditions, since it is blocked by the beta-blocker propranolol administration. In the first experiment we showed that the brain cortex is involved in the thermogenesis induced by LH lesion and this effect is mediated by SNS, since bilateral functional decortication induced by cortical-spreading depression (CSD) impaired the increase of O2 consumption to the same extent as administration of propranolol. In the second experiment the role played by the cerebral cortex on thermogenesis in rats during voluntary overeating of "cafeteria" diet and in control rats was investigated. Cafeteria rats showed a significantly higher colonic temperature, brown adipose tissue temperature (Tbat), and rate of O2 consumption than control animals. CSD led to a significant decrease of Tbat and O2 consumption in cafeteria rats but not in controls. On the basis of the results obtained in the two experiments, the possibility that the cerebral cortex could be involved in the metabolic responses for reduction of body weight to the "set-point" is hypothesized.