196 research outputs found
HSV-replication defective based vector as vaccines against Rotavirus infections
No full textBackground: Rotaviruses (RVs) are the most important cause of acute gastroenteritis in humans and animals. These viruses cause diarrheal disease primarily in the young, but infection and disease in older children and adults can occur, resulting in more than 600 000 deaths per annum, mainly in developing countries.
RV particle is nonenveloped, with the viral genome of 11 segments of double-stranded RNA surrounded by three concentric protein layers: the outer layer is composed of VP (Viral Protein) 7 and VP4, the intermediate layer is formed by VP6, and the innermost layer is composed by VP2.
Studies of natural infection in children indicated that primary infections can protect against severe disease upon secondary infection. For these reasons the most frequent approach to Rotavirus vaccine design was based on the use of attenuated animal or human Rotavirus strains, orally delivered to mimic natural infection. However, several major drawbacks have affected the development and application of these live Rotavirus-based oral vaccines, including the withdrawal from market of one of these vaccines because of adverse effects (gut intussusception). Moreover Rotaviruses are constantly and rapidly evolving, due both to their segmented genomes and to the fact that their genes can reassort between strains coinfecting a same host, and this fact is an important threat to the rational of using live attenuated Rotavirus strains as vaccines, both because these strains can revert to more virulent phenotypes and because these vaccine strains are being released to the nature in the faeces of the inoculated person, raising a considerable ecological concern. It is therefore critical to develop alternatives to this classical approach, both to generate a deeper understanding and to explore the potential of novel vaccination strategies.
Aim: the goal of this project is the production of a collection of plasmids and HSV-1-based vectors expressing Rotavirus antigens as tool for research on Rotaviruses and the development of innovative genetic vaccines to fight against Rotavirus, based on the use of replication-defective HSV-1 vectors.
Methods: the Rotavirus genes (vp7, vp4, vp6 and vp2) were subcloned first in basic plasmids (pcDNA Hygro 3.1+ or 3.1-, or in pBSSK) then in plasmids that have HSV specific loci sequences (pB41, pB5, pgJHE), where the Rotavirus cassettes were inserted between these Herpes sequences in order to recombine them into the viral genomes. Homologous recombinations have been carried out using standard calcium phosphate transfection.
The recombinant backbones used (T0ZGFP, THZ and S0ZgJGFP) are replication-defective HSV-1 viral vectors that has low toxicity due to the deletion of one or more immediate early genes essential for viral replication; in addition they contain marker genes (gfp gene or lacZ gene) useful for the identification of the recombinant backbone which has integrated the transgene in its genome.
Transfection and isolation of the recombinant viruses were performed in modified Vero cells (7b or E5), capable of providing the essential immediate early gene products in trans. The recombinant viruses containing the Rotavirus transgenes were identified by isolation of a clear plaque phenotype for GFP under the fluorescent microscope or after X-gal staining.
All the recombinant viruses have been purified by three rounds of limiting dilution and the presence of the transgenes was verified by Southern blot analysis. The protein expression was evaluated by immunofluorescence and Western blot techniques with specific mono/polyclonal antibodies.
Main results: a lot of plasmids containing Rotavirus genes of human, mice or simian origin and a large set of HSV-1-based vectors expressing Rotavirus proteins were constructed. The expression of VP6, VP2 and VP7 Rotavirus proteins (of murine or simian origin, under ICP0 promoter or HCMV promoter control, and in different loci of HSV genome) was visible in Western blot analysis. Moreover, nanotubes formed by VP6 protein were detected after immunofluorescence assays.
Conclusions: plasmids and recombinant HSV-vectors carrying Rotavirus genes represent a tool to improve the research on Rotavirus and to elucidate the features of individual Rotavirus antigens. HSV-1-based vectors expressing Rotavirus antigens could represent an attractive alternative strategy to current vaccines. In addition, the possibility to cross recombinant HSV-vectors carrying Rotavirus genes in different loci of HSV could enhance the immune response by the construction of virus-like particles (VLPs); VLPs are a highly effective type of subunit vaccines that mimic the overall structure of virus particles without any requirement that they contain infectious genetic material
Malignant Pleural Mesothelioma: Management and Role of Radiation Therapy
No full textMalignant pleural mesothelioma is a neoplasm derived from the mesothelial surfaces of the pleura. There are
tree different mesothelioma types: Epithelioid Mesothelioma; Sarcomatoid; Biphasic /Mixed Mesothelioma. Patients
with mesothelioma have a poor prognosis with a median survival ranging from 6 to 18 months depending on the
stage of the disease at the time of diagnosis
CONCOMITANT MALIGNANT MESOTHELIOMA OF THE PLEURA, PERITONEUM AND TUNICA VAGINALIS TESTIS
No full textFamilial pleural malignant mesothelioma: clustering in three sisters and one cousin
No full textMalignant mesothelioma is associated with asbestos, yet its occurrence in genetically-related individuals suggests a role of host predisposition. We have identified a cluster of pleural malignant mesothelioma in three sisters and one cousin (male). The women had worked in the same confectionery shop as pastry cooks and/or pastry shop assistants; the use of an asbestos-insulated oven was the putative source of exposure. The man had occupational exposure as a heating system installation worker. The family history reported malignant cancers in first-degree (larynx, brother; pleura and lung, mother), second-degree (lung, aunt and uncle) and third-degree (lung, cousin) relatives. The study reveals the potential existence of the mesothelioma risk among pastry cooks and highlights the fact that inherited factors may be involved in the development of this tumour. (C) 1998 Elsevier Science Inland Ltd. All rights reserved
Beclomethasone dipropionate, formoterol fumarate and glycopyrronium bromide: Synergy of triple combination therapy on human airway smooth muscle ex vivo
No full textCombining inhaled corticosteroids (ICSs), long-acting β2 -adrenoceptor agonists (LABAs) and long-acting muscarinic antagonists (LAMAs) is recommended to treat severe forms of asthma and chronic obstructive pulmonary disease (COPD). Clinical benefits have been demonstrated for ICS/LABA/LAMA combinations. This study characterized the interaction between the ICS beclomethasone dipropionate, the LABA formoterol fumarate and the LAMA glycopyrronium bromide in human airways
Pharmacological investigation on the anti-oxidant and anti-inflammatory activity of N-acetylcysteine in an ex vivo model of COPD exacerbation
No full textOxidative stress is recognized to be one of predisposing factor in the pathogenesis of COPD. The oxidant/antioxidant imbalance is significantly pronounced in patients with COPD exacerbation. N-acetylcysteine (NAC) seems to be able to reduce COPD exacerbations by modulating the oxidative stress in addition to its well-known mucolytic activity, but there are discordant findings on the actual anti-oxidant activity of NAC
Beclomethasone dipropionate and formoterol fumarate synergistically interact in hyperresponsive medium bronchi and small airways
No full textCorticosteroids increase the expression of β2-adrenoceptors (β2-ARs) and protect them against down-regulation. Conversely, β2-AR agonists improve the anti-inflammatory action of corticosteroids. Nevertheless, it is still uncertain whether adding a long-acting β2-AR agonist (LABA) to an inhaled corticosteroid (ICS) results in an additive effect, or there is true synergy. Therefore, the aim of this study was to pharmacologically characterize the interaction between the ICS beclomethasone diproprionate (BDP) and the LABA formoterol fumarate (FF) in a validated human ex vivo model of bronchial asthma
Beclomethasone dipropionate and sodium cromoglycate protect against airway hyperresponsiveness in a human ex vivo model of cow's milk aspiration
No full textBackground: Recurrent cow's milk (CM) aspiration is often associated with gastroesophageal reflux in infants and toddlers and it seems to be implicated in the etiology of different inflammatory lung disorders. This study aimed to investigate ex vivo the impact of CM aspiration on human airways and whether treatment with beclomethasone dipropionate (BDP) or sodium cromoglycate (SCG) may prevent the potential CM-induced airway hyperresponsiveness (AHR). Methods: Human isolated bronchi were contracted by electrical field stimulation (EFS10Hz) to mimic the contractile tone induced by the parasympathetic activity and challenged with CM, fat/lactose-free CM, or human breast milk (HM). The effect of pre-treatment with beclomethasone dipropionate (BDP) and sodium cromoglycate (SCG) was also investigated on the AHR induced by CM. Results: After a 60 min-challenge with CM 1:10 v/v and fat/lactose-free CM 1:10 v/v, ASM significantly (P < 0.05) increased compared to control (+67.04 ± 17.08% and +77.91 ± 1.34%, respectively), a condition that remained stable for 150 min post-treatment, whereas HM did not alter ASM contractility. BDP 1 μM and 10 μM significantly (P < 0.05) reduced the AHR elicited by CM (-52.49 ± 10.97% and -66.98 ± 7.90%, respectively vs. control). At the same manner, SCG 1 μM and 10 μM significantly (P < 0.05) inhibited the CM-induced AHR (-59.03 ± 9.24% and -73.52 ± 7.41%, respectively vs. control). Conclusion: CM induces AHR in human ASM by eliciting an increased parasympathetic contractile response. Preventive treatment with nebulized SCG may be indicated in infants or toddlers fed with CM, rather than with BDP due to a superior safety profile
A rare case of oral multisystem Langerhans cell histiocytosis
No full textLangerhans cell histiocytosis (LCH) is a rare disorder characterized by high proliferation of Langerhans dendritic cells. LCH is a solitary or multifocal disease that primarily involves bone tissue and often affects children and young men. A 29 years-old Caucasian man was referred to the Oral Surgery Unit of George Eastman Hospital - Umberto I teaching hospital, with third degree mobility of teeth belonging to second, third and fourth quadrant. Panoramic radiograph showed multiple radiolucent areas with well demarcated borders on the right and left site of the mandible and on the left site of the maxilla. Extractions of compromised teeth and biopsy of the osteolytic tissue were performed. The final diagnosis of multisystem Langerhans cell histiocytosis of the soft and hard tissues of the oral cavity was made. The patient was sent to the Hematology department of Umberto I Teaching Hospital of ?Sapienza? ? University of Rome for the proper treatment. The present case of rare multisystem LCH involving oral hard and soft tissues shows the strong importance of better investigate, with appropriate additional exams, initial shifty symptoms that could lead to a misdiagnosis
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