86,606 research outputs found
Analisi biometrica e ciclo di maturità sessuale in Mytilus galloprovincialis Lmk in allevamento a Grignano (Nota preliminare).
No full textCaratteristiche dell'accrescimento e del periodo riproduttivo in un allevamento sperimentale di Mytilus galloprovincialis Lam.
No full textRecovery of EDTA and metal precipitation from soil flushing solutions
No full textThis work studies the effectiveness of a process proposed for the recovery of ethylenediaminetetraacetic acid (EDTA) and metal precipitation from soil flushing solutions. Two series of experimental tests were carried out on two samples of a soil artificially contaminated with copper or lead. The metals were extracted from the soil by flushing with a 0.05M aqueous solution of EDTA sodium salt (E-Na2). Cu or Pb extraction efficiencies of about 95 and 98% were observed, respectively. The two extracted solutions were then treated to obtain EDTA recovery and metal precipitation from the aqueous solution. EDTA recovery was achieved in two steps. An initial evaporation treatment lead to reduce the solution volume by about 75%. This was followed by the acidification of the residue solution, which precipitated more than 93% of the used EDTA. The precipitated EDTA was removed by filtration, and was suitable for reuse after adding an alkaline agent. Metal precipitation from the filtered solution was performed using two different methods: an almost total (99.5%) Pb precipitation in alkaline conditions was achieved after complex destabilization through the sequential addition of ferric ions and sodium phosphate, while 93.6% copper precipitation was achieved with ferrous sulfate as a destabilization agent
N-terminal fatty acids of NEFmut are required for the CD8+ t-cell immunogenicity of in vivo engineered extracellular vesicles
No full textWe recently described a cytotoxic CD8+ T lymphocyte (CTL) vaccine platform based on the intramuscular (i.m.) injection of DNA eukaryotic vectors expressing antigens of interest fused at the C-terminus of HIV-1 Nefmut, i.e., a functionally defective mutant that is incorporated at quite high levels into exosomes/extracellular vesicles (EVs). This system has been proven to elicit strong CTL immunity against a plethora of both viral and tumor antigens, as well as inhibit both transplantable and orthotopic tumors in mice. However, a number of open issues remain regarding the underlying mechanism. Here we provide evidence that hindering the uploading into EVs of Nefmut-derived products by removing the Nefmut N-terminal fatty acids leads to a dramatic reduction of the downstream antigen-specific CD8+ T-cell activation after i.m. injection of DNA vectors in mice. This result formally demonstrates that the generation of engineered EVs is part of the mechanism underlying the in vivo induced CD8+ T-cell immunogenicity. Gaining new insights on the EV-based vaccine platform can be relevant in view of its possible translation into the clinic to counteract both chronic and acute infections as well as tumors
Antitumor HPV E7-specific CTL activity elicited by in vivo engineered exosomes produced through DNA inoculation
No full textPaola Di Bonito,1 Chiara Chiozzini,2 Claudia Arenaccio,2 Simona Anticoli,2 Francesco Manfredi,2 Eleonora Olivetta,2 Flavia Ferrantelli,2 Emiliana Falcone,3 Anna Ruggieri,3 Maurizio Federico2 1Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy; 2National AIDS Center, Istituto Superiore di Sanità, Rome, Italy; 3Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità, Rome, Italy Abstract: We recently proved that exosomes engineered in vitro to deliver high amounts of HPV E7 upon fusion with the Nefmut exosome-anchoring protein elicit an efficient anti-E7 cytotoxic T lymphocyte immune response. However, in view of a potential clinic application of this finding, our exosome-based immunization strategy was faced with possible technical difficulties including industrial manufacturing, cost of production, and storage. To overcome these hurdles, we designed an as yet unproven exosome-based immunization strategy relying on delivery by intramuscular inoculation of a DNA vector expressing Nefmut fused with HPV E7. In this way, we predicted that the expression of the Nefmut/E7 vector in muscle cells would result in a continuous source of endogenous (ie, produced by the inoculated host) engineered exosomes able to induce an E7-specific immune response. To assess this hypothesis, we first demonstrated that the injection of a Nefmut/green fluorescent protein-expressing vector led to the release of fluorescent exosomes, as detected in plasma of inoculated mice. Then, we observed that mice inoculated intramuscularly with a vector expressing Nefmut/E7 developed a CD8+ T-cell immune response against both Nef and E7. Conversely, no CD8+ T-cell responses were detected upon injection of vectors expressing either the wild-type Nef isoform of E7 alone, most likely a consequence of their inefficient exosome incorporation. The production of immunogenic exosomes in the DNA-injected mice was formally demonstrated by the E7-specific CD8+ T-cell immune response we detected in mice inoculated with exosomes isolated from plasma of mice inoculated with the Nefmut/E7 vector. Finally, we provide evidence that the injection of Nefmut/E7 DNA led to the generation of effective antigen-specific cytotoxic T lymphocytes whose activity was likely part of the potent, therapeutic antitumor effect we observed in mice implanted with TC-1 tumor cells. In summary, we established a novel method to generate immunogenic exosomes in vivo by the intramuscular inoculation of DNA vectors expressing the exosome-anchoring protein Nefmut and its derivatives. Keywords: nanovesicles, cytotoxic T lymphocytes, HIV-1 Nef, DNA vector
Effect on physiological parameters and anaesthetic dose requirement of isoflurane when tramadol given as a continuous rate infusion vs a single intravenous bolus injection during ovariohysterectomy in dogs
No full textBackground
Tramadol produces a significant reduction in both sevoflurane and isoflurane minimum alveolar
concentrations in dogs under experimental conditions. This study aims to compare the
effects of tramadol administered as a constant rate infusion (CRI) with those of tramadol
administered as a single intravenous bolus on physiological parameters and isoflurane
requirements in dogs undergoing ovariohysterectomy.
Methods
In this study, forty female dogs undergoing ovariohysterectomy were enrolled. The bitches
were anesthetized with 5 mg/kg of tiletamine/zolazepam combined with 0.05 mg/kg of acepromazine
intravenously. Anesthesia was maintained with isoflurane delivered in 100% oxygen.
The group A (n = 20) received tramadol 4 mg/kg in a single intravenous bolus, whereas
the group B (n = 20) received tramadol 1.5 mg/kg in an intravenous bolus followed by tramadol
2.6 mg/kg/h as a CRI. The following parameters were recorded: heart rate, respiratory
rate, non-invasive blood pressure, body temperature, EtCO2, SpO2 and inspired and
expired concentrations of isoflurane. Parameter measurements were performed from prepreedication
(baseline) to skin suturing.
Results
The dogs were healthy subjects that demonstrated no abnormalities on laboratory investigations.
Significant tachycardia was recorded after administration of tiletamine/zolazepam
combined with acepromazine in both groups. Heart rate decreased after intubation but
remained significantly higher compared to baseline values in both groups. Systolic blood
pressure significantly decreased in both groups but the recorded values were within thephysiological range. Mild reduction in body temperature was recorded in both groups. SpO2
and EtCO2 remained within the physiological range. Isoflurane requirement was significantly
lower in the group B compared to the group A. Transient twitching was recorded in two dogs
belonging to the group A after tramadol administration.
Conclusions
Compared to tramadol given as a single intravenous bolus injection during ovariohysterectomy
in dogs, tramadol administered as a CRI reduces isoflurane requirements in dogs anesthetized
with tiletamine/zolazepam combined with acepromazine. Both tramadol given as a
CRI and a single intravenous bolus injection, induce decrease in heart rate, respiratory rate
and in body temperature but the values of these parameters remain within physiological
range in dogs undergoing ovariohysterectomy
Extracellular vesicle-mediated intercellular communication in HIV-1 infection and its role in the reservoir maintenance
No full textHIV-1 infection is efficiently controlled by combination anti-retroviral therapy (cART). However, despite preventing disease progression, cART does not eradicate virus infection which persists in a latent form for an individual's lifetime. The latent reservoir comprises memory CD4+ T lymphocytes, macrophages, and dendritic cells; however, for the most part, the reservoir is generated by virus entry into activated CD4+ T lymphocytes committed to return to a resting state, even though resting CD4+ T lymphocytes can be latently infected as well. The HIV-1 reservoir is not recognized by the immune system, is quite stable, and has the potential to re-seed systemic viremia upon cART interruption. Viral rebound can occur even after a long period of cART interruption. This event is most likely a consequence of the extended half-life of the HIV-1 reservoir, the maintenance of which is not clearly understood. Several recent studies have identified extracellular vesicles (EVs) as a driving force contributing to HIV-1 reservoir preservation. In this review, we discuss recent findings in the field of EV/HIV-1 interplay, and then propose a mechanism through which EVs may contribute to HIV-1 persistence despite cART. Understanding the basis of the HIV-1 reservoir maintenance continues to be a matter of great relevance in view of the limitations of current strategies aimed at HIV-1 eradication
Vaccines based on the native HIV Tat protein and on the combination of Tat and the structural HIV protein variant DeltaV2 Env.
No full textThe promising results obtained with the HIV-1 Tat-based vaccines in mice, monkeys and humans, a better understanding of Tat immunomodulatory functions, as well as evidence that vaccination with trimeric V2 loop-deleted HIV-1 Env induces cross-clade neutralizing antibodies led to the rational design of a novel vaccine based on the combination of Tat and V2-deleted Env
On the morphology of Anisakis pegreffii: a comparative analysis of three microscopic techniques used to build a new parasite atlas
No full textBACKGROUND: Human anisakidosis is a parasitic anthropozoonosis caused by larval
nematodes of the family Anisakidae. Here, we report a detailed description of the
morphology of Anisakis pegreffii third-stage larva performed using a conventional light
and confocal microscopy, and scanning electron microscopy (SEM) that provide a
basis for both phenotypic studies and genetic mutations.
METHODS: The collected larvae from fish were morphologically identified as Anisakis
larvae Type I, and they were characterized by PCR-RFLP to identify the Anisakis
pegreffii specie. Using NC5/NC2 primers, ribosomal genomic regions ITS1, 5.8 SrRNA
and ITS2 of DNA were amplified and PCR products were sequenced. Fifteen larvae
belonging to Anisakis pegreffii were fixed, sectioned, and examined with a light and
confocal microscope and by SEM.
RESULTS: In our studies, have been acquired detailed ultrastructural images, which
have been integrated with those derived from the dissection of the parasite, obtained
with light and confocal microscopy. The structural and ultrastructural images
concerning the third stage larvae of Anisakis pegreffii have been studied, analyzed and
compared among them. The derived overall view has allowed detecting new interesting
details of a well-known parasite and has been schematically showed.
CONCLUSIONS: The aim of this study is to furnish an updated atlas of Anisakis
pegreffii. Confocal microscopy, as well as the light and electron microscopy have
played a pivotal role in the accumulation of new scientific data regarding the anatomical
structures of this nematode. This work is the result of one year of engagement by the
Authors and the outcome is a comprehensive atlas on Anisakis pegreffii microscopy
Variations on the Author
No full text“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
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