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Further to a previous publication by the European Council of Legal Medicine (ECLM) concerning on-site forensic and medico-legal scene and corpse investigation, this publication provides guidance for forensic medical specialists, pathologists and, where present, coroners' activity at a scene of death inspection and to harmonize the procedures for a correct search, detection, collection, sampling and storage of all elements which may be useful as evidence, and ensure documentation of all these steps. This ECLM's inspection form provides a checklist to be used on-site for the investigation of a corpse present at a crime or suspicious death scene. It permits the collection of all relevant data not only for the pathologist, but also for forensic anthropologists, odontologists, geneticists, entomologists and toxicologists, thus supporting a collaborative work approach. Detailed instructions for the completion of forms are provided.
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Entomología , Medicina Legal , Antropología , Cadáver , Medicina Legal/métodos , Patologia Forense , HumanosRESUMEN
In the last years, change in multiple sclerosis (MS) therapeutic scenario has highlighted the need for an improved doctor-patient communication in advance of treatment initiation in order to allow patient's empowerment in the decision-making process. AIMS: The aims of our project were to review the strategies used by Italian MS specialists to inform patients about treatment options and to design a multicentre shared document that homogenizes the information about disease-modifying treatment (DMTs) and the procedure of taking informed consent in clinical practice. RESULTS: The new resource, obtained by consensus among 31 neurologists from 27 MS Centres in Italy with the supervision of a medico-legal advisor, received the aegis of Italian Neurological Society (SIN) and constitutes a step toward a standardized decision process around DMTs in MS.
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Consentimiento Informado , Esclerosis Múltiple , Consenso , Humanos , Italia , Esclerosis Múltiple/terapia , Relaciones Médico-PacienteRESUMEN
Chromosomes are key players of cell physiology, their dynamics provides valuable information about its physical organization. In both prokaryotes and eukaryotes, the short-time motion of chromosomal loci has been described with a Rouse model in a simple or viscoelastic medium. However, little emphasis has been put on the influence of the folded organization of chromosomes on the local dynamics. Clearly, stress propagation, and thus dynamics, must be affected by such organization, but a theory allowing us to extract such information from data, e.g., on two-point correlations, is lacking. Here, we describe a theoretical framework able to answer this general polymer dynamics question. We provide a scaling analysis of the stress-propagation time between two loci at a given arclength distance along the chromosomal coordinate. The results suggest a precise way to assess folding information from the dynamical coupling of chromosome segments. Additionally, we realize this framework in a specific model of a polymer whose long-range interactions are designed to make it fold in a fractal way and immersed in a medium characterized by subdiffusive fractional Langevin motion with a tunable scaling exponent. This allows us to derive explicit analytical expressions for the correlation functions.
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Cromosomas/química , Cromosomas/genética , Modelos Químicos , Modelos Genéticos , Escherichia coli/genética , Escherichia coli/ultraestructura , Fractales , Humanos , ViscosidadRESUMEN
The ototoxic effect of the exposure to styrene is evaluated, also in the presence of simultaneous exposure to noise, using otoacoustic emissions as biomarkers of mild cochlear damage. Transient-evoked and distortion product otoacoustic emissions were recorded and analyzed in a sample of workers (15 subjects) exposed to styrene and noise in a fiberglass manufacturing facility and in a control group of 13 non-exposed subjects. Individual exposure monitoring of the airborne styrene concentrations was performed, as well as biological monitoring, based on the urinary concentration of two styrene metabolites, the Mandelic and Phenylglyoxylic acids. Noise exposure was evaluated using wearable phonometers, and hearing loss with pure tone audiometry. Due to their different job tasks, one group of workers was exposed to high noise and low styrene levels, another group to higher styrene levels, close to the limit of 20 ppm, and to low noise levels. A significant negative correlation was found between the otoacoustic emission levels and the concentration of the styrene urinary metabolites. Otoacoustic emissions, and particularly distortion products, were able to discriminate the exposed workers from the controls, providing also a rough estimate of the slope of the dose-response relation between otoacoustic levels and styrene exposure.
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Contaminantes Ocupacionales del Aire/efectos adversos , Cóclea/efectos de los fármacos , Pérdida Auditiva Provocada por Ruido/etiología , Ruido/efectos adversos , Emisiones Otoacústicas Espontáneas/efectos de los fármacos , Estireno/efectos adversos , Adulto , Contaminantes Ocupacionales del Aire/orina , Audiometría de Tonos Puros , Biomarcadores/orina , Biotransformación , Estudios de Casos y Controles , Cromatografía Líquida de Alta Presión , Cóclea/lesiones , Cóclea/fisiopatología , Relación Dosis-Respuesta a Droga , Monitoreo del Ambiente/métodos , Femenino , Cromatografía de Gases y Espectrometría de Masas , Pérdida Auditiva Provocada por Ruido/diagnóstico , Pérdida Auditiva Provocada por Ruido/fisiopatología , Humanos , Perfil Laboral , Masculino , Persona de Mediana Edad , Exposición Profesional/efectos adversos , Salud Laboral , Saliva/metabolismo , Estireno/orina , Espectrometría de Masas en TándemRESUMEN
Modified vaccinia virus Ankara (MVA) is an attenuated poxvirus strain, currently under evaluation as a vaccine vector in various clinical settings. It has been reported that human dendritic cells (DCs) mature after infection with MVA, but reports on the functionality of DCs have so far been controversial. In this work, we studied the phenotype and functionality of MVA-infected DCs. As previously reported, we found that human monocyte-derived DCs upregulated CD86 and HLA-DR in response to MVA infection. Moreover, infected DCs produced a broad array of chemokines and cytokines and were able to activate and induce gamma interferon (IFN-γ) production both in CD4(+) and in CD8(+) allogeneic T cells and in specific autologous peripheral blood lymphocytes (PBLs). Analysis of DC maturation following infection with a recombinant green fluorescent protein (GFP)-expressing MVA revealed that upregulation of CD86 expression was mainly observed in GFP(neg) (bystander) cells. While GFP(pos) (infected) DCs produced tumor necrosis factor alpha (TNF-α), they were unable to produce CXCL10 and were less efficient at inducing IFN-γ production in CEF-specific autologous PBLs. Maturation of bystander DCs could be achieved by incubation with supernatant from infected cultures or with apoptotic infected cells. Type I IFNs were partially responsible for the induction of CXCL10 on bystander DCs. Our findings demonstrate for the first time that, in MVA-infected DC cultures, the leading role with respect to functionality and maturation characteristics is achieved by the bystander DCs.
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Células Dendríticas/inmunología , Virus Vaccinia/inmunología , Antígeno B7-2/análisis , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Células Cultivadas , Citocinas/metabolismo , Células Dendríticas/química , Antígenos HLA-DR/análisis , Humanos , Interferón gamma/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Modern deep neural networks (DNNs) represent a formidable challenge for theorists: according to the commonly accepted probabilistic framework that describes their performance, these architectures should overfit due to the huge number of parameters to train, but in practice they do not. Here we employ results from replica mean field theory to compute the generalization gap of machine learning models with quenched features, in the teacher-student scenario and for regression problems with quadratic loss function. Notably, this framework includes the case of DNNs where the last layer is optimized given a specific realization of the remaining weights. We show how these results-combined with ideas from statistical learning theory-provide a stringent asymptotic upper bound on the generalization gap of fully trained DNN as a function of the size of the dataset P. In particular, in the limit of large P and N_{out} (where N_{out} is the size of the last layer) and N_{out}âªP, the generalization gap approaches zero faster than 2N_{out}/P, for any choice of both architecture and teacher function. Notably, this result greatly improves existing bounds from statistical learning theory. We test our predictions on a broad range of architectures, from toy fully connected neural networks with few hidden layers to state-of-the-art deep convolutional neural networks.
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This study is aimed at testing the cochlear functionality using otoacoustic emissions, analyzing the synergistic effects of simultaneous exposure to noise and organic solvents EBTx on workers of a glass-reinforced plastic products factory. Exposure to organic solvents was assessed using ambiental measurements and evaluation of the salivary concentration. Biological monitoring was performed evaluating the urinary concentration of the styrene metabolites. Statistical analysis shows that otoacoustic tests can discriminate between different exposure groups. Significant differences were found between controls and subjects exposed to high styrene and low noise levels, showing the ototoxic effect (at sub-clinical level) of the styrene exposure.
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Cóclea/efectos de los fármacos , Cóclea/fisiopatología , Ruido en el Ambiente de Trabajo/efectos adversos , Exposición Profesional/efectos adversos , Estireno/efectos adversos , Adulto , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
In critical systems, the effect of a localized perturbation affects points that are arbitrarily far from the perturbation location. In this paper, we study the effect of localized perturbations on the solution of the random dimer problem in two dimensions. By means of an accurate numerical analysis, we show that a local perturbation of the optimal covering induces an excitation whose size is extensive with finite probability. We compute the fractal dimension of the excitations and scaling exponents. In particular, excitations in random dimer problems on nonbipartite lattices have the same statistical properties of domain walls in spin glass. Excitations produced in bipartite lattices, instead, are compatible with a loop-erased self-avoiding random walk process. In both cases, we find evidence of conformal invariance of the excitations that is compatible with SLE_{κ} with parameter κ depending on the bipartiteness of the underlying lattice only.
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BACKGROUND: DNA bridging promoted by the H-NS protein, combined with the compaction induced by cellular crowding, plays a major role in the structuring of the E. coli genome. However, only few studies consider the effects of the physical interplay of these two factors in a controlled environment. METHODS: We apply a single molecule technique (Magnetic Tweezers) to study the nanomechanics of compaction and folding kinetics of a 6 kb DNA fragment, induced by H-NS bridging and/or PEG crowding. RESULTS: In the presence of H-NS alone, the DNA shows a step-wise collapse driven by the formation of multiple bridges, and little variations in the H-NS concentration-dependent unfolding force. Conversely, the DNA collapse force observed with PEG was highly dependent on the volume fraction of the crowding agent. The two limit cases were interpreted considering the models of loop formation in a pulled chain and pulling of an equilibrium globule respectively. CONCLUSIONS: We observed an evident cooperative effect between H-NS activity and the depletion of forces induced by PEG. GENERAL SIGNIFICANCE: Our data suggest a double role for H-NS in enhancing compaction while forming specific loops, which could be crucial in vivo for defining specific mesoscale domains in chromosomal regions in response to environmental changes.
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ADN Bacteriano/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Proteínas Fimbrias/metabolismo , Polietilenglicoles/metabolismo , ADN Bacteriano/química , Escherichia coli/química , Fenómenos Magnéticos , Conformación de Ácido NucleicoRESUMEN
Neuromuscular diseases (NMD) may affect respiratory muscles, leading to respiratory failure. Studies show that long-term noninvasive mechanical ventilation (NIV) improves symptoms, gas exchange, quality of life and survival. NIV improved these parameters in muscular dystrophies and also in patients with amyotrophic lateral sclerosis without severe bulbar dysfunction. NIV should be started at the onset of nocturnal hypoventilation. In selected cases, NIV may be simpler, better accepted by patients and cheaper than invasive mechanical ventilation, but it cannot be used as an alternative. Tracheostomy may be preferred by patients unable to protect their airways and wishing to survive as long as possible, or by ventilator-dependent patients. Glossopharyngeal breathing consists of taking air and propelling it into the lungs. Chest percussions and vibrations can help to mobilise airway secretions but they cannot substitute coughing. Manually assisted coughing requires substantial lung inflation through air stacking or deep lung insufflation, followed by an abdominal thrust with open glottis. The combination of mechanical in-exsufflation with an abdominal thrust is a mechanically assisted cough. In conclusion, recent advances in respiratory care of NMD have improved prognosis and many caregivers have changed from a traditional noninterventional to a more aggressive, supportive approach.
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Enfermedades Neuromusculares/terapia , Neumología/métodos , Adulto , Esclerosis Amiotrófica Lateral/fisiopatología , Esclerosis Amiotrófica Lateral/terapia , Tos , Humanos , Insuflación , Distrofias Musculares/fisiopatología , Distrofias Musculares/terapia , Enfermedades Neuromusculares/fisiopatología , Neumología/tendencias , Calidad de Vida , Respiración Artificial , Insuficiencia Respiratoria/fisiopatología , Insuficiencia Respiratoria/terapia , Sistema Respiratorio , Terapia Respiratoria/métodos , Traqueostomía/métodosRESUMEN
Hygienists are interested in hexavalent chromium due to its genotoxic and carcinogenic effect on humans. The use of products containing hexavalent chromium is decreasing in many industrial fields because of the substitution with less-toxic compounds. In the aeronautical industry, however, the chromate are added to primer paint as a corrosion inhibitor of aircrafts surfaces: so hexavalent chromium compounds are available in many primers with a composition ranging from 10% to 13%. The application of these primers by using electrostatic guns potentially exposes painting and coating workers at high concentrations of aerosols containing Cr(VI). The aim of the present study is the evaluation of professional exposure to hexavalent chromium during aircraft painting, by adopting both environmental personal sampling and biological monitoring. To valuate workers exposure levels the personal measurements results have been compared with the exposure limit values (TLV-TWA) and the urinary chromium contents with the biological exposure indices (IBE). Moreover the strategy of coupling environmental sampling with biological monitoring seems to be a useful instrument to measure the validity of the individual protection devices.
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Contaminación del Aire/análisis , Carcinógenos Ambientales/análisis , Cromo/orina , Industrias , Exposición Profesional/análisis , Aeronaves , HumanosRESUMEN
An optimal vaccine against leishmaniasis should elicit parasite specific CD4+ and cytotoxic CD8+ T cells. In this investigation, we described a prime/boost immunization approach based on DNA and on poxvirus vectors (Western Reserve, WR, and the highly attenuated modified vaccinia virus Ankara, MVA), both expressing the LACK antigen of Leishmania infantum, that triggers different levels of specific CD8+ T cell responses and protection (reduction in lesion size and parasitemia) against L. major infection in mice. A prime/boost vaccination with DNA-LACK/MVA-LACK elicits higher CD8+ T cell responses than a similar protocol with the replication competent VV-LACK. Both CD4+ and CD8+ T cells were induced by DNA-LACK/MVA-LACK immunization. The levels of IFN-gamma and TNF-alpha secreting CD8+ T cells were higher in splenocytes from DNA-LACK/MVA-LACK than in DNA-LACK/VV-LACK immunized animals. Moreover, protection against L. major was significantly higher in DNA-LACK/MVA-LACK than in DNA-LACK/VV-LACK immunized animals when boosted with the same virus dose, and correlated with high levels of IFN-gamma and TNF-alpha secreting CD8+ T cells. In DNA-LACK/MVA-LACK vaccinated animals, the extent of lesion size reduction ranged from 65 to 92% and this protection was maintained for at least 17 weeks after challenge with the parasite. These findings demonstrate that in heterologous prime/boost immunization approaches, the protocol DNA-LACK/MVA-LACK is superior to DNA-LACK/VV-LACK in triggering specific CD8+ T cell immune responses and in conferring protection against cutaneous leishmaniasis. Thus, MVA-LACK is a safe and efficient vector for vaccination against leishmaniasis.
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Antígenos de Protozoos/inmunología , Leishmaniasis Cutánea/inmunología , Leishmaniasis Cutánea/prevención & control , Proteínas Protozoarias/inmunología , Vacunas Antiprotozoos/inmunología , Virus Vaccinia , Animales , Antígenos de Protozoos/genética , ADN Protozoario , Esquema de Medicación , Femenino , Inmunización Secundaria , Interferón gamma/metabolismo , Leishmania major , Leishmaniasis Cutánea/patología , Ratones , Ratones Endogámicos BALB C , Proteínas Protozoarias/genética , Vacunas Antiprotozoos/efectos adversos , Vacunas Antiprotozoos/genética , Linfocitos T/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Vacunas Sintéticas/efectos adversos , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunologíaRESUMEN
The aim of this study was to verify whether different living conditions of Polychaeta are correlated with morphological and functional differences in the organization of the integument. For this purpose, we decided to study the epidermis of Timarete filigera, a non-tubicolous polychaete. With this objective in mind, we have identified the various cellular types responsible for mucous secretion in the epidermis of this species and defined the histochemical composition of the mucus produced by different types of gland cells. Three types of gland cells have been identified by histochemical and ultrastructural studies in the epidermis of this polychaete. The histochemistry was carried out using standard techniques and peroxidase-labelled lectins. In type 1 cells, the secretory granules contain neutral glycoproteins with glucosidic residues of GalNAc, Galbeta 1,3 GalNAc, glucosidic and/or mannosidic residues. In type 2 cells, the secretory granules contain acid glycoproteins mainly sulphated with glucosidic residues of GalNAc, Galbeta 1,3 GalNAc, glucosidic and/or mannosidic residues, and some terminal sialic acid. In type 3 cells, the residual granules have the same chemical composition as that of granules present in type 2 cells. The secretion of these glandular mucous cells consists of mainly sulphated acidic glycoproteins and GAG resistant to testis jaluronidase. In these cells, the residual granules have the same chemical composition as that of their secretion. The heterogeneity of mucus composition may be correlated with its different functions.
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Epidermis/ultraestructura , Células Epiteliales/metabolismo , Glándulas Exocrinas/ultraestructura , Moco/metabolismo , Poliquetos/ultraestructura , Animales , Epidermis/metabolismo , Glándulas Exocrinas/metabolismo , Glucósidos/química , Glucósidos/metabolismo , Glicoproteínas/química , Glicoproteínas/metabolismo , Glicosaminoglicanos/química , Glicosaminoglicanos/metabolismo , Histocitoquímica , Lectinas , Microscopía Electrónica de Transmisión , Moco/química , Ácido N-Acetilneuramínico/química , Ácido N-Acetilneuramínico/metabolismo , Poliquetos/metabolismo , Vesículas Secretoras/química , Vesículas Secretoras/metabolismo , Vesículas Secretoras/ultraestructura , Especificidad de la EspecieRESUMEN
The use of chemicals during restoration practices involves emissions of toxics depending on both the amount of original products used and the specific techniques applied. Restorers perform a great variety of techniques by using not standardized operative procedures: moreover, they are exposed to mixtures and very rarely to single chemicals. In this study we evaluated workers' exposure to solvent mixture which were generated during restoration of archaeological metal handcraft. Exposure to low levels of twelve organic compounds has been experimented inside a public restoration laboratory: based on the data presented in this article we assume that inside restoration workplaces a diffused indoor contamination can take place. The indoor pollution can generate a risk of an uncontrolled exposure to volatile mixtures: therefore the emissions of solvents during restoration practices has to be avoided even if they cause a low level of exposure.
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Contaminación del Aire Interior/análisis , Arte , Exposición Profesional/análisis , Solventes/análisis , Humanos , Factores de Riesgo , VolatilizaciónRESUMEN
Protocols of immunization based on the DNA prime/vaccinia virus (VV) boost regime with recombinants expressing relevant antigens have been shown to elicit protection against a variety of pathogens in animal model systems, and various phase I clinical trials have been initiated with this vaccination approach. We have previously shown that mice immunized with a DNA vector expressing p36/LACK of Leishmania infantum followed by a booster with VVp36/LACK induced significant protection against Leishmania major infection. To further improve this protocol of immunization, here we investigated whether the cytokines interleukin-12 (IL-12) and IL-18 could enhance protection against L. major infection in BALB/c mice. We found that priming with DNA vectors expressing p36/LACK and either IL-12 or IL-18, followed by a booster with a VV recombinant expressing the same L. infantum LACK antigen, elicit a higher cellular immune response than by using the same protocol in the absence of the cytokines. The cytokine IL-12 triggered a higher number of IFN-gamma-secreting cells specific for p36 protein than IL-18. When immunized animals were challenged with promastigotes, the highest protection against L. major infection was observed in animals primed with DNAp36 + DNA IL-12 + DNA IL-18 and boosted with VVp36. This protection correlated with a Th1 type of immune response. Our findings revealed that in prime/booster protocols, co-expressing IL-12 and IL-18 during priming is an efficient approach to protect against leishmaniasis. This combined prime/booster immunization regime could have wide use in fighting against parasitic and other infectious diseases.
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Antígenos de Protozoos , Inmunización , Interleucina-12/inmunología , Interleucina-18/inmunología , Leishmaniasis Cutánea/prevención & control , Proteínas Protozoarias/inmunología , Virus Vaccinia/genética , Animales , Anticuerpos Antiprotozoarios/sangre , Femenino , Vectores Genéticos , Esquemas de Inmunización , Inmunización Secundaria , Interleucina-12/genética , Interleucina-12/metabolismo , Interleucina-18/genética , Interleucina-18/metabolismo , Leishmania infantum/inmunología , Leishmania major/patogenicidad , Leishmaniasis Cutánea/inmunología , Leishmaniasis Cutánea/parasitología , Ratones , Ratones Endogámicos BALB C , Proteínas Protozoarias/genética , Vacunas Antiprotozoos/administración & dosificación , Vacunas Antiprotozoos/inmunología , Células TH1/inmunología , Vacunas de ADN/inmunologíaRESUMEN
A main goal of the industrialized world is the development of effective vaccines to control infectious diseases with major health and socio-economic impact. Current understanding of the immune response triggered during infection with pathogens causing malaria, hepatitis C and AIDS emphasizes the importance of cytotoxic T lymphocytes (CTLs) in combating these infections. This has led to the development of new vaccination strategies, some of which are in phase I/II clinical trials. Promising strategies of vaccination are based on highly attenuated viral vectors, such as Vaccinia virus (VV) in combination with heterologous like vectors naked DNA, referred to as priming/booster vaccination. While these immunization schedules increased the production of specific CTLs, there is a need to further expand the CD8+T cell population to control an infection. Among molecules that play a significant role in the modulation of the CTL response is the cytokine IL-12. Immunoregulation by IL-12 is of central importance in cell-mediated immunity (CMI) against those pathogens and tumors that are controlled by cell-mediated mechanisms, supported by Thl cells. The use of this cytokine in combination with highly immunogenic VV-derived vectors is a promising system for development of future vaccination schedules. In this review, we summarize recent data on the use of IL-12 in vaccination procedures, as well as undesired side-effects of the cytokine that can be overcome by accurate use of dose, route and time-window administration of IL-12 encoding vectors. Results described here indicate that VV IL-12-mediated enhancement of the specific CMI response against a model antigen HIV-1 env was time- and dose-dependent and that the antigen and the cytokine could be expresed from two different rVVs modulating the doses of the vectors and allowing for enhancement of a specific CMI response. Moreover, the use of IL-12 during DNA prime/VV boost regimens enhanced the specific anti-HIV-1 env cellular response 20 times compared to that generated after a single rVVenv inoculation. Variables such as: a) dose of the cytokine applied, b) time of its administration and c) routes of inoculation play a critical role in the final outcome of the response. The findings presented here can be extended to other antigens, suggesting that immunomodulatory cytokines can be useful in the development of the future vaccines against numerous infectious diseases and tumors.
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Adyuvantes Inmunológicos , Inmunidad Celular , Inmunización Secundaria , Interleucina-12/inmunología , Vacunas de ADN/inmunología , Vacunas Virales/inmunología , Animales , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Citocinas/metabolismo , Proteínas gp160 de Envoltorio del VIH/genética , Proteínas gp160 de Envoltorio del VIH/inmunología , VIH-1/inmunología , Humanos , Interleucina-12/fisiología , Linfocitos T Citotóxicos/inmunología , Células TH1/inmunología , Células TH1/metabolismo , Virus Vaccinia/genética , Virus Vaccinia/inmunologíaRESUMEN
Temperature-sensitive mutants of Streptococcus pneumoniae were isolated after chemical mutagenesis. Intranasal immunization with temperature-sensitive mutant J/3 induced higher levels of circulating antibody than those obtained after immunization with the heat-killed parental wild type. Moreover, local immunization with mutant J/3 induced high levels of anti-S. pneumoniae IgG and IgA in the lower respiratory tract, whereas only moderate IgG (and no IgA) antibodies were detected in lung lavage fluids from mice immunized intranasally with the heat-killed strain.
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Anticuerpos Antibacterianos/biosíntesis , Cápsulas Bacterianas/inmunología , Infecciones Neumocócicas/inmunología , Administración Intranasal , Animales , Anticuerpos Antibacterianos/sangre , Formación de Anticuerpos , Líquido del Lavado Bronquioalveolar/inmunología , Inmunidad Activa , Inmunoglobulina A/análisis , Inmunoglobulina G/análisis , Pulmón/inmunología , Ratones , Ratones Endogámicos , Membrana Mucosa/inmunología , Mutagénesis , Infecciones Neumocócicas/genéticaRESUMEN
A temperature-sensitive filamentation (fts) Salmonella typhimurium mutant was isolated after transposon mutagenesis with mini-Tn 10dTc. The mutant was unable to form colonies after 20 h incubation at 37 degrees C on LB agar. Colonies appeared, however, after longer incubation at the restrictive temperature. Filamentation affected only part of the bacterial population. Rapid mapping using Mu dP22 hybrid phages revealed that the mutation, ftsD220, lies within minutes 68.5 and 73.6 on the genetic map. Further analysis revealed that the ftsD220 mapped at min 73 and that it is linked to cysG (6%) and to aroB (39%). Complementation tests suggested that the ftsD220 mutation is not homologous to a Escherichia coli ftsH mutation.
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Fimbrias Bacterianas/genética , Salmonella typhimurium/genética , Mapeo Cromosómico , Elementos Transponibles de ADN , Microscopía de Contraste de Fase , Mutagénesis Insercional , Fenotipo , Salmonella typhimurium/ultraestructura , Temperatura , Transducción GenéticaRESUMEN
Female mice were immunized by the intramammary route with live-attenuated Staphylococcus aureus according to different schedules and challenged with virulent S. aureus. Immunization in late pregnancy or early lactation induced a significant decrease (P <0.05) in the number of S. aureus CFU recovered from glands after the challenge and a significant increase (P <0.05) in the levels of milk and serum specific IgG and IgA antibodies. Mice immunized before pregnancy were not protected from S. aureus challenge. Immunization did not increase the number of somatic cells in milk when compared with control mice. Protection from S. aureus intramammary infection may be achieved if mice are locally immunized during late pregnancy or early lactation.
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Vacunas Bacterianas , Mastitis/prevención & control , Complicaciones Infecciosas del Embarazo/prevención & control , Infecciones Estafilocócicas/prevención & control , Staphylococcus aureus/inmunología , Animales , Anticuerpos Antibacterianos/análisis , Anticuerpos Antibacterianos/sangre , Vacunas Bacterianas/administración & dosificación , Vacunas Bacterianas/inmunología , Femenino , Esquemas de Inmunización , Inmunoglobulina A/análisis , Inmunoglobulina A/sangre , Inmunoglobulina G/análisis , Inmunoglobulina G/sangre , Glándulas Mamarias Animales/inmunología , Glándulas Mamarias Animales/microbiología , Glándulas Mamarias Animales/patología , Mastitis/inmunología , Ratones , Leche/inmunología , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Infecciones Estafilocócicas/inmunología , Vacunación , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/inmunologíaRESUMEN
The persistence of Salmonella enteritidis temperature-sensitive (ts) mutants of different phenotypes in Peyer's patches (PP) and the spleen, and their immunogenicity after intragastric (i.g.) and peroral (p.o.) administration to mice was investigated. After p.o. administration the ts mutant C/2/2 colonized PP, but was not recovered from the spleen. After i.g. administration the ts mutant E/1/3 colonized both the spleen and PP for at least 2 weeks. Mutant C/2/2 persisted in PP up to 8 days but was not found in the spleen. Mutant H/2/26, although it poorly colonized the PP, was recovered from the spleen up to day 15 after i.g. administration. Immunization with E/1/3 by either the i.g. or the p.o. routes protected mice from challenge with 100 LD50 of the virulent wild-type (wt) strain. Immunization with either C/2/2 or H/2/26 did not confer protection. The three ts mutants induced the production of local IgA after i.g. administration regardless of their protective capacity.