Characterizing the distribution of a STING agonist and its metabolites in mouse liver by MALDI imaging mass spectrometry.

A STING (stimulator of interferon genes) agonist GSK3996915 under investigation in early discovery for hepatitis B was orally dosed to a mouse model for understanding the parent drug distribution in liver, the target organ. MALDI imaging mass spectrometry (IMS) was used to quantify the distribution of GSK3996915 in liver collected from mice administered a single oral dose at 90 mg/kg. GSK3996915 was detected with a zonal distribution localized in the portal triad and highly concentrated in the main bile ducts, indicating clearance through biliary excretion. High spatial resolution imaging showed the distribution of the parent drug localized to the cellular populations in the sinusoids including the Kupffer cells. Additionally, a series of drug-related metabolites were observed to be localized in the central zones of the liver. These results exemplify the potential of utilizing MALDI IMS for measuring not only quantitative drug distribution and target exposure, but also drug metabolism and elimination in a single suite of experiments. Significance Statement An integrated imaging approach utilizing MALDI IMS, immunohistochemistry (IHC), and histology was used to measure MALDI IMS complemented with other imaging techniques such as immunohistochemistry addressed the question of target exposure at the cellular level. Localized quantification of the parent drug in the target organ and identificaitonidentification of potential metabolites in the context of tissue histology were also achieved in one experimental suite to support characterization of pharmacokinetic properties of the drug in the early discovery stage.

T Cell- and Monocyte-Specific RNA-Sequencing Analysis in Septic and Nonseptic Critically Ill Patients and in Patients with Cancer.

Sepsis is characterized as life-threatening organ dysfunction caused by a dysregulated host immune response to infection. The purpose of this investigation was to determine the differential effect of sepsis on innate versus adaptive immunity, in humans, by examining RNA expression in specific immune cell subsets, including monocytes/macrophages and CD4 and CD8 T cells. A second aim was to determine immunosuppressive mechanisms operative in sepsis that might be amenable to immunotherapy. Finally, we examined RNA expression in peripheral cells from critically ill nonseptic patients and from cancer patients to compare the unique immune response in these disorders with that occurring in sepsis. Monocytes, CD4 T cells, and CD8 T cells from septic patients, critically ill nonseptic patients, patients with metastatic colon cancer, and healthy controls were analyzed by RNA sequencing. Sepsis induced a marked phenotypic shift toward downregulation of multiple immune response pathways in monocytes suggesting that impaired innate immunity may be fundamental to the immunosuppression that characterizes the disorder. In the sepsis cohort, there was a much more pronounced effect on gene transcription in CD4 T cells than in CD8 T cells. Potential mediators of sepsis-induced immunosuppression included Arg-1, SOCS-1, and SOCS-3, which were highly upregulated in multiple cell types. Multiple negative costimulatory molecules, including TIGIT, Lag-3, PD-1, and CTLA-4, were also highly upregulated in sepsis. Although cancer had much more profound effects on gene transcription in CD8 T cells, common immunosuppressive mechanisms were present in all disorders, suggesting that immunoadjuvant therapies that are effective in one disease may also be efficacious in the others.

Author Correction: Design of amidobenzimidazole STING receptor agonists with systemic activity.

Change history: In this Letter, author Ana Puhl was inadvertently omitted; this error has been corrected online.An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Design of amidobenzimidazole STING receptor agonists with systemic activity.

Stimulator of interferon genes (STING) is a receptor in the endoplasmic reticulum that propagates innate immune sensing of cytosolic pathogen-derived and self DNA1. The development of compounds that modulate STING has recently been the focus of intense research for the treatment of cancer and infectious diseases and as vaccine adjuvants2. To our knowledge, current efforts are focused on the development of modified cyclic dinucleotides that mimic the endogenous STING ligand cGAMP; these have progressed into clinical trials in patients with solid accessible tumours amenable to intratumoral delivery3. Here we report the discovery of a small molecule STING agonist that is not a cyclic dinucleotide and is systemically efficacious for treating tumours in mice. We developed a linking strategy to synergize the effect of two symmetry-related amidobenzimidazole (ABZI)-based compounds to create linked ABZIs (diABZIs) with enhanced binding to STING and cellular function. Intravenous administration of a diABZI STING agonist to immunocompetent mice with established syngeneic colon tumours elicited strong anti-tumour activity, with complete and lasting regression of tumours. Our findings represent a milestone in the rapidly growing field of immune-modifying cancer therapies.

T Regulatory Cells Support Plasma Cell Populations in the Bone Marrow.

Long-lived plasma cells (PCs) in the bone marrow (BM) are a critical source of antibodies after infection or vaccination, but questions remain about the factors that control PCs. We found that systemic infection alters the BM, greatly reducing PCs and regulatory T (Treg) cells, a population that contributes to immune privilege in the BM. The use of intravital imaging revealed that BM Treg cells display a distinct behavior characterized by sustained co-localization with PCs and CD11c-YFP+ cells. Gene expression profiling indicated that BM Treg cells express high levels of Treg effector molecules, and CTLA-4 deletion in these cells resulted in elevated PCs. Furthermore, preservation of Treg cells during systemic infection prevents PC loss, while Treg cell depletion in uninfected mice reduced PC populations. These studies suggest a role for Treg cells in PC biology and provide a potential target for the modulation of PCs during vaccine-induced humoral responses or autoimmunity.

Inflammasome Priming in Sterile Inflammatory Disease.

The inflammasome is a cytoplasmic protein complex that processes interleukins (IL)-1ß and IL-18, and drives a form of cell death known as pyroptosis. Oligomerization of this complex is actually the second step of activation, and a priming step must occur first. This involves transcriptional upregulation of pro-IL-1ß, inflammasome sensor NLRP3, or the non-canonical inflammasome sensor caspase-11. An additional aspect of priming is the post-translational modification of particular inflammasome constituents. Priming is typically accomplished in vitro using a microbial Toll-like receptor (TLR) ligand. However, it is now clear that inflammasomes are activated during the progression of sterile inflammatory diseases such as atherosclerosis, metabolic disease, and neuroinflammatory disorders. Therefore, it is time to consider the endogenous factors and mechanisms that may prime the inflammasome in these conditions.

Distinct modes of antigen presentation promote the formation, differentiation, and activity of foxp3+ regulatory T cells in vivo.

How the formation and activity of CD4(+)Foxp3(+) regulatory T cells (Tregs) are shaped by TCR recognition of the diverse array of peptide:MHC complexes that can be generated from self-antigens and/or foreign Ags in vivo remains poorly understood. We show that a self-peptide with low (but not high) stimulatory potency promotes thymic Treg formation and can induce conventional CD4(+) T cells in the periphery to become Tregs that express different levels of the transcription factor Helios according to anatomical location. When Tregs generated in response to this self-peptide subsequently encountered the same peptide derived instead from influenza virus in the lung-draining lymph nodes of infected mice, they proliferated, acquired a T-bet(+)CXCR3(+) phenotype, and suppressed the antiviral effector T cell response in the lungs. However, these self-antigen-selected Tregs were unable to suppress the antiviral immune response based on recognition of the peptide as a self-antigen rather than a viral Ag. Notably, when expressed in a more immunostimulatory form, the self-peptide inhibited the formation of T-bet(+)CXCR3(+) Tregs in response to viral Ag, and Ag-expressing B cells from these mice induced Treg division without upregulation of CXCR3. These studies show that a weakly immunostimulatory self-peptide can induce thymic and peripheral Foxp3(+) Treg formation but is unable to activate self-antigen-selected Tregs to modulate an antiviral immune response. Moreover, a strongly immunostimulatory self-peptide expressed by B cells induced Tregs to proliferate without acquiring an effector phenotype that allows trafficking from the draining lymph node to the lungs and, thereby, prevented the Tregs from suppressing the antiviral immune response.

Coinfection with Streptococcus pneumoniae modulates the B cell response to influenza virus.

Pathogen-specific antibodies (Abs) protect against respiratory infection with influenza A virus (IAV) and Streptococcus pneumoniae and are the basis of effective vaccines. Sequential or overlapping coinfections with both pathogens are common, yet the impact of coinfection on the generation and maintenance of Ab responses is largely unknown. We report here that the B cell response to IAV is altered in mice coinfected with IAV and S. pneumoniae and that this response differs, depending on the order of pathogen exposure. In mice exposed to S. pneumoniae prior to IAV, the initial virus-specific germinal center (GC) B cell response is significantly enhanced in the lung-draining mediastinal lymph node and spleen, and there is an increase in CD4(+) T follicular helper (TFH) cell numbers. In contrast, secondary S. pneumoniae infection exaggerates early antiviral antibody-secreting cell formation, and at later times, levels of GCs, TFH cells, and antiviral serum IgG are elevated. Mice exposed to S. pneumoniae prior to IAV do not maintain the initially robust GC response in secondary lymphoid organs and exhibit reduced antiviral serum IgG with diminished virus neutralization activity a month after infection. Our data suggest that the history of pathogen exposures can critically affect the generation of protective antiviral Abs and may partially explain the differential susceptibility to and disease outcomes from IAV infection in humans. Importance: Respiratory tract coinfections, specifically those involving influenza A viruses and Streptococcus pneumoniae, remain a top global health burden. We sought to determine how S. pneumoniae coinfection modulates the B cell immune response to influenza virus since antibodies are key mediators of protection.

Pneumolysin expression by streptococcus pneumoniae protects colonized mice from influenza virus-induced disease.

The response to influenza virus (IAV) infection and severity of disease is highly variable in humans. We hypothesized that one factor contributing to this variability is the presence of specific respiratory tract (RT) microbes. One such microbe is Streptococcus pneumoniae (Sp) that is carried asymptomatically in the RT of many humans. In a mouse co-infection model we found that in contrast to secondary bacterial infection that exacerbates disease, Sp colonization 10 days prior to IAV protects from virus-induced morbidity and lung pathology. Using mutant Sp strains, we identified a critical role for the bacterial virulence factor pneumolysin (PLY) in mediating this protection. Colonization with the PLY-sufficient Sp strain induces expression of the immune-suppressive enzyme arginase 1 in alveolar macrophages (aMø) and correlates with attenuated recruitment and function of pulmonary inflammatory cells. Our study demonstrates a novel role for PLY in Sp-mediated protection by maintaining aMø as "gatekeepers" against virus-induced immunopathology.

The transcription factor Foxp1 is a critical negative regulator of the differentiation of follicular helper T cells.

CD4(+) follicular helper T cells (T(FH) cells) are essential for germinal center (GC) responses and long-lived antibody responses. Here we report that naive CD4(+) T cells deficient in the transcription factor Foxp1 'preferentially' differentiated into T(FH) cells, which resulted in substantially enhanced GC and antibody responses. We found that Foxp1 used both constitutive Foxp1A and Foxp1D induced by stimulation of the T cell antigen receptor (TCR) to inhibit the generation of T(FH) cells. Mechanistically, Foxp1 directly and negatively regulated interleukin 21 (IL-21); Foxp1 also dampened expression of the costimulatory molecule ICOS and its downstream signaling at early stages of T cell activation, which rendered Foxp1-deficient CD4(+) T cells partially resistant to blockade of the ICOS ligand (ICOSL) during T(FH) cell development. Our findings demonstrate that Foxp1 is a critical negative regulator of T(FH) cell differentiation.

A promiscuous lipid-binding protein diversifies the subcellular sites of toll-like receptor signal transduction.

The Toll-like receptors (TLRs) of the innate immune system are unusual in that individual family members are located on different organelles, yet most activate a common signaling pathway important for host defense. It remains unclear how this common signaling pathway can be activated from multiple subcellular locations. Here, we report that, in response to natural activators of innate immunity, the sorting adaptor TIRAP regulates TLR signaling from the plasma membrane and endosomes. TLR signaling from both locations triggers the TIRAP-dependent assembly of the myddosome, a protein complex that controls proinflammatory cytokine expression. The actions of TIRAP depend on the promiscuity of its phosphoinositide-binding domain. Different lipid targets of this domain direct TIRAP to different organelles, allowing it to survey multiple compartments for the presence of activated TLRs. These data establish how promiscuity, rather than specificity, can be a beneficial means of diversifying the subcellular sites of innate immune signal transduction.

Estudio y análisis médico-legal de las muertes en privación de libertad en Málaga (2004-2012) / Study and forensic analysis of deaths in custody in Malaga, Spain (2004-2012)

Se presenta un estudio sobre todos los casos de muertes en privación de libertad ocurridos en la provincia de Málaga durante los años 2004-2012, que fueron 41. En cuanto al origen y la causa de la muerte, la mayoría fueron accidentales (46,3%, incluyendo reacción adversa a drogas), seguidas de suicidios (24,3%), muertes de origen natural (21,9%) y homicidios (7,3%). En el estudio se detalla también la edad, el sexo, la nacionalidad, el consumo de tóxicos y el lugar del suceso. Finalmente se exponen cuatro interesantes casos: dos accidentes, un suicidio y un homicidio (AU)

A study of all cases of deaths in custody occurred in the province of Malaga, Spain, during the years 2004 to 2012 is presented. These were 41. As for the origin and etiology of death, most were accidents 46.3% (including adverse drug reaction), followed by suicides (24.3%), natural deaths (21.9%) and homicides (7.3%). In the study; age, sex, national consumption of toxic and location of occurrence is detailed. Finally four interesting cases are discussed: twoaccidents, one suicide and a homicide (AU)

Cooperativity between CD8+ T cells, non-neutralizing antibodies, and alveolar macrophages is important for heterosubtypic influenza virus immunity.

Seasonal epidemics of influenza virus result in ∼36,000 deaths annually in the United States. Current vaccines against influenza virus elicit an antibody response specific for the envelope glycoproteins. However, high mutation rates result in the emergence of new viral serotypes, which elude neutralization by preexisting antibodies. T lymphocytes have been reported to be capable of mediating heterosubtypic protection through recognition of internal, more conserved, influenza virus proteins. Here, we demonstrate using a recombinant influenza virus expressing the LCMV GP33-41 epitope that influenza virus-specific CD8+ T cells and virus-specific non-neutralizing antibodies each are relatively ineffective at conferring heterosubtypic protective immunity alone. However, when combined virus-specific CD8 T cells and non-neutralizing antibodies cooperatively elicit robust protective immunity. This synergistic improvement in protective immunity is dependent, at least in part, on alveolar macrophages and/or other lung phagocytes. Overall, our studies suggest that an influenza vaccine capable of eliciting both CD8+ T cells and antibodies specific for highly conserved influenza proteins may be able to provide heterosubtypic protection in humans, and act as the basis for a potential "universal" vaccine.

Microsatellite marker-based identification and genetic relationships of olive cultivars from the Extremadura region of Spain.

Seventy-seven olive accessions corresponding to 25 cultivars from the Extremadura region of Spain were studied using four microsatellite or SSR markers in order to fingerprint them, and evaluate genetic similarity and relationships between local and introduced olive cultivars. The number of alleles per locus ranged from 4 to 8, with a mean of 6.25 alleles per primer pair (a total of 25 alleles). The observed heterozygosity ranged from 0.58 to 0.95, while the expected heterozygosity varied between 0.68 and 0.83. The polymorphism information content values ranged from 0.63 to 0.79. The mean polymorphism information content value of 0.70 for the SSR loci provided sufficient discriminating ability to evaluate the genetic diversity among the cultivars. The SSR data allowed unequivocal identification of all the cultivars; a combination of three SSR markers was sufficient to discriminate all 25 olive cultivars. A dendrogram was prepared, using the unweighted pair-group method with arithmetic mean clustering algorithm; it depicted the pattern of relationships between the cultivars. Most of the local cultivars grouped according to their geographic origin. No clear clustering trends were observed when the morphological traits of fruit endocarps or fruit use of cultivars were employed as analysis criteria. We conclude that there is a high level of variability among local olive cultivars from the Extremadura region at both the morphological and molecular levels; these data should be useful for identifying and distinguishing local germplasm.

Vaccination with M2e-based multiple antigenic peptides: characterization of the B cell response and protection efficacy in inbred and outbred mice.

BACKGROUND: The extracellular domain of the influenza A virus protein matrix protein 2 (M2e) is remarkably conserved between various human isolates and thus is a viable target antigen for a universal influenza vaccine. With the goal of inducing protection in multiple mouse haplotypes, M2e-based multiple antigenic peptides (M2e-MAP) were synthesized to contain promiscuous T helper determinants from the Plasmodium falciparum circumsporozoite protein, the hepatitis B virus antigen and the influenza virus hemagglutinin. Here, we investigated the nature of the M2e-MAP-induced B cell response in terms of the distribution of antibody (Ab) secreting cells (ASCs) and Ab isotypes, and tested the protective efficacy in various mouse strains. METHODOLOGY/PRINCIPAL FINDINGS: Immunization of BALB/c mice with M2e-MAPs together with potent adjuvants, CpG 1826 oligonucleotides (ODN) and cholera toxin (CT) elicited high M2e-specific serum Ab titers that protected mice against viral challenge. Subcutaneous (s.c.) and intranasal (i.n.) delivery of M2e-MAPs resulted in the induction of IgG in serum and airway secretions, however only i.n. immunization induced anti-M2e IgA ASCs locally in the lungs, correlating with M2-specific IgA in the bronchio-alveolar lavage (BAL). Interestingly, both routes of vaccination resulted in equal protection against viral challenge. Moreover, M2e-MAPs induced cross-reactive and protective responses to diverse M2e peptides and variant influenza viruses. However, in contrast to BALB/c mice, immunization of other inbred and outbred mouse strains did not induce protective Abs. This correlated with a defect in T cell but not B cell responsiveness to the M2e-MAPs. CONCLUSION/SIGNIFICANCE: Anti-M2e Abs induced by M2e-MAPs are highly cross-reactive and can mediate protection to variant viruses. Although synthetic MAPs are promising designs for vaccines, future constructs will need to be optimized for use in the genetically heterogeneous human population.

Consumo de alcohol, tabaco, cannabis y otras sustancias psicoactivas en estudiantes de la Universidad de Málaga / Consumption of alcohol, tobacco, cannabis and other psychoactive substances among University of Malaga students

Objetivos. Conocer la prevalencia de consumo de sustancias psicoactivas en los estudiantes de la Universidad de Málaga, patrones de consumo, accesibilidad y riesgo percibido, con el fin de obtener información que permita diseñar intervenciones dirigidas a la prevención y modificación de actitudes. Material y métodos. Cuestionario anónimo y autocumplimentado administrado a una muestra representativa (n = 1.121) de la población de estudiantes (bienio 2007-2009). Resultados. Las sustancias más consumidas en el último mes fueron alcohol (64,6%) y cannabis (9%), con una prevalencia de consumo para todas las sustancias por debajo de la media en la población de referencia, excepto para el alcohol, donde se iguala. El consumo de tabaco diario (11,2%) es muy inferior al de la población general, aunque similar al de otras universidades. Perciben como conductas de consumo menos arriesgadas el consumo de alcohol (23-24,5%) y el consumo esporádico de cannabis (20,6%), que consideran la droga ilícita más accesible. La mayoría de las sustancias presentan un uso recreativo y son consumidas en grupo. No se aprecian diferencias por razón de género en la percepción del riesgo para ninguna de las conductas, aun cuando el patrón de consumo sí varía en función del sexo. Conclusiones. La prevalencia de consumo para alcohol y cannabis junto a la percepción del riesgo asociado al uso de las sustancias psicoactivas estudiadas y otros datos obtenidos en este estudio podrían contribuir a definir intervenciones dirigidas a la prevención y modificación de actitudes, así como al diseño de futuras investigaciones en este colectivo (AU)

Objectives. To find out the prevalence of psychoactive substance use among students in Malaga University, and their consumption patterns, accessibility and risk awareness associated with their use in order to obtain information to design interventions for prevention and behavioural changes. Material and methods. An anonymous self-administered questionnaire were used in a representative sample of students population (n=1,121, period 2007-2009). Results. The substances most consumed in the last month were alcohol (64.6%) and cannabis (9%), with a prevalence of consumption for all the substances under the average in the population of reference, except for alcohol that remains equal. The consumption of daily smoked tobacco (11.2%) is lower than that of the general population, although similar to other universities. Student perceive low risk associated with alcohol consumption (23-24.5%) as well as sporadic consumption of cannabis (20.6%) which is consider the most accessible illicit drug. Most substances show a recreational use and are usually consumed in groups. There are not gender differences in risk awareness for any of the substances, although the consumption patterns for the majority of substances depend on the gender. Conclusions. Prevalence of both alcohol and cannabis consumption as well as risk awareness associated with psychoactive substance use, and other findings from this study could address the development of interventions for prevention, behavioural changes and the design of new studies in this group (AU)

Protective antiviral antibody responses in a mouse model of influenza virus infection require TACI.

Antiviral Abs, for example those produced in response to influenza virus infection, are critical for virus neutralization and defense against secondary infection. While the half-life of Abs is short, Ab titers can last a lifetime due to a subset of the Ab-secreting cells (ASCs) that is long lived. However, the mechanisms governing ASC longevity are poorly understood. Here, we have identified a critical role for extrinsic cytokine signals in the survival of respiratory tract ASCs in a mouse model of influenza infection. Irradiation of mice at various time points after influenza virus infection markedly diminished numbers of lung ASCs, suggesting that they are short-lived and require extrinsic factors in order to persist. Neutralization of the TNF superfamily cytokines B lymphocyte stimulator (BLyS; also known as BAFF) and a proliferation-inducing ligand (APRIL) reduced numbers of antiviral ASCs in the lungs and bone marrow, whereas ASCs in the spleen and lung-draining lymph node were surprisingly unaffected. Mice deficient in transmembrane activator and calcium-modulator and cyclophilin ligand interactor (TACI), a receptor for BLyS and APRIL, mounted an initial antiviral B cell response similar to that generated in WT mice but failed to sustain protective Ab titers in the airways and serum, leading to increased susceptibility to secondary viral challenge. These studies highlight the importance of TACI signaling for the maintenance of ASCs and protection against influenza virus infection.

Agitación psicomotriz e implicaciones médico forenses / Psychomotor agitation and forensic implications

Observaciones llevadas a cabo durante los últimos 15 años por expertos forenses han dado lugar al reconocimiento de un nuevo síndrome que puede llevar a la muerte súbita e inesperada. La muerte por agitación psicomotriz generalmente se acompaña por un procedimiento de restricción llevado a cabo por fuerzas y cuerpos de seguridad del estado, cuerpos de seguridad privada o personal hospitalario. Ello conlleva que dicho fallecimiento sea catalogado dentro del grupo de muertes en privación de libertad, muertes en custodia o deaths in custody. La frecuencia de este cuadro, llamativo por las circunstancias extrañas en las que se produce, es cada vez mayor ya que uno de sus principales factores de riesgo como es el consumo de drogas, fundamentalmente la cocaína, aumenta de forma alarmante en la sociedad de nuestros días.Por tanto, partiendo de la hipótesis de que la agitación psicomotriz de origen tóxico o psiquiátrico puede acabar en la muerte del sujeto tras una retención prolongada así como que algunos de los sujetos que sufren un cuadro de agitación psicomotriz son ingresados en el hospital durante o tras dicho episodio, nuestro objetivo ha sido realizar un estudio descriptivo utilizando para ello una muestra de 31 casos que requirieron atención médica en el Hospital de Poniente entre los años 2003 y 2005. Esta muestra la dividimos en dos subgrupos: • Sujetos con agitación psicomotriz y análisis cualitativo de tóxicos positivo y, • Sujetos con agitación psicomotriz y análisis cualitativo de tóxicos negativo. Con esta división en dos grupos realizamos un estudio comparativo entre ambos analizando estadísticamente diferentes variables con el fin de determinar la influencia de las drogas psicoactivas en los cuadros de agitación psicomotriz. Los resultados obtenidos ponen de manifiesto la falta de datos en las historias clínicas, el escaso número de estudios relacionados con el tema en cuestión y la ausencia de diferencias estadísticamente significativas entre los pacientes con agitación psicomotriz y consumo de tóxicos de los que no han consumido recientemente

Observations carried out by forensic experts over the last 15 years lead us to the recognition of a new syndrome that can take to a sudden and unexpected death. Death caused by psychomotor agitation is generally accompanied by a restriction procedure carried out by security forces, private security or hospital staff. It entails that this death can be classified within the group of deaths in deprivation of liberty or deaths in custody. The frequency of this condition, remarkable for the strange circumstances in which it occurs, is growing as one of their major risk factors such as drugs, primarily cocaine, increases dramatically in today's society. Therefore, based on the hypothesis that psychomotor agitation due to toxic or psychiatric origin can end in the death of the subject after a prolonged retention as well as some of the subjects suffering from psychomotor agitation are admitted to hospital during or after this episode, our goal was to perform a descriptive study using a sample of 31 cases requiring medical attention in the Poniente Hospital between 2003 and 2005. This sample was divided into two subgroups: • Patients with psychomotor agitation and positive toxic qualitative analysis and, • Patients with psychomotor agitation and negative toxic qualitative analysis. A comparative study was carried out between the two different groups, statistically analyzing different variables to determine the influence of psychoactive drugs in the state of psychomotor agitation. Results highlight the lack of data in medical records, the small numbers of studies related to the topic and no statistically significant differences between patients with psychomotor agitation and toxic consumption of those who have not recently consumed

Plasmacytoid dendritic cells are dispensable during primary influenza virus infection.

Plasmacytoid dendritic cells (pDC) are thought to be pivotal in the first line of defense against viral infections. Although previous studies have suggested that pDC regulate the immune response against respiratory syncytial virus, their role in pulmonary infection with influenza virus has remained unclear. Using mice with GFP-tagged pDC, we observed a marked increase in pDC numbers in the lung airways 3 days after intranasal infection with influenza virus A/PR/8/34. To further investigate their potential involvement in the disease, we made use of pDC-deficient IkarosL/L mice. In the absence of pDC, the recruitment of T cells to the bronchoalveolar space was delayed, which could be reversed by the adoptive transfer of pDC before infection. Surprisingly, however, when compared with wild-type animals, IkarosL/L mice revealed a similar course of disease, as determined by weight loss, viral titers, levels of neutralizing Ab, and lung pathology. Moreover, the activation and differentiation of influenza-specific CD8+ effector T cells was unaltered in the absence of pDC, as was the generation of CD8+ memory T cells. Taken together, our study suggests that pDC regulate the accumulation of T cells in the bronchoalveolar space during early influenza virus infection, but are dispensable for the control of this disease.

A tomato peroxidase involved in the synthesis of lignin and suberin.

The last step in the synthesis of lignin and suberin has been proposed to be catalyzed by peroxidases, although other proteins may also be involved. To determine which peroxidases are involved in the synthesis of lignin and suberin, five peroxidases from tomato (Lycopersicon esculentum) roots, representing the majority of the peroxidase activity in this organ, have been partially purified and characterized kinetically. The purified peroxidases with isoelectric point (pI) values of 3.6 and 9.6 showed the highest catalytic efficiency when the substrate used was syringaldazine, an analog of lignin monomer. Using a combination of transgenic expression and antibody recognition, we now show that the peroxidase pI 9.6 is probably encoded by TPX1, a tomato peroxidase gene we have previously isolated. In situ RNA hybridization revealed that TPX1 expression is restricted to cells undergoing synthesis of lignin and suberin. Salt stress has been reported to induce the synthesis of lignin and/or suberin. This stress applied to tomato caused changes in the expression pattern of TPX1 and induced the TPX1 protein. We propose that the TPX1 product is involved in the synthesis of lignin and suberin.