Long-term antigen-specific immune response by an oncolytic adenovirus encoding SP-SA-E7-4-1BBL in HPV-16 cancer model.

BACKGROUND: To describe an oncolytic adenovirus (OAd) encoding SP-SA-E7-4-1BBL that is capable of inducing tumor regression in therapeutic assays. Herein, we tested whether the antitumor effect is given by the induction of a tumor-specific immune response, as well as the minimum dose needed to elicit antitumor protection and monitor the OAd biodistribution over time. METHODS AND RESULTS: C57BL/6 mice (n = 5) per group were immunized twice with OAds encoding SP-SA-E7-4-1BBL, SA-E7-4-1BBL, or SP-SA-4-1BBL and challenged with TC-1 cancer cells. The DNA construct SP-SA-E7-4-1BBL was employed as a control via biolistic or PBS injection. Groups without tumor development at 47 days were rechallenged with TC-1 cells, and follow-up lasted until day 90. The minimum dose of OAd to induce the antitumor effect was established by immunization using serial dilution doses. The cytometry bead assay and the ELISpot assay were used to evaluate cytokine release in response to ex vivo antigenic stimulation. The distribution profile of the OAd vaccine was evaluated in the different organs by histological, immunohistochemical and qPCR analyses. The OAd SP-SA-E7-4-1BBL-immunized mice did not develop tumors even in a rechallenge. A protective antitumor effect was observed from a dose that is one hundredth of most reports of adenoviral vaccines. Immunization with OAd increases Interferon-gamma-producing cells in response to antigen stimulation. OAd was detected in tumors over time, with significant morphological changes, contrary to nontumor tissues. CONCLUSIONS: The OAd SP-SA-E7-4-1BBL vaccine confers a prophylactic, safe, long-lasting, and antigen-dependent antitumor effect mediated by a Th1 antitumor immune response.

Implementation of a Novel Prehospital Clinical Decision Tool and ECG Transmission for STEMI Significantly Reduces Door-to-Balloon Time and Sex-Based Disparities.

BACKGROUND: An important method employed to reduce door to balloon time (DTBT) for ST segment elevation Myocardial Infarctions (STEMIs) is a prehospital MI alert. The purpose of this retrospective study was to examine the effects of an educational intervention using a novel decision support method of STEMI notification and prehospital electrocardiogram (ECG) transmission on DTBT. METHODS: An ongoing database (April 4, 2000 - present) is maintained to track STEMI alerts. In 2007, an MI alert program began; emergency medicine physicians could activate a "prehospital MI alert". In October 2015, modems were purchased for Emergency Medical Services personnel to transmit ECGs. There was concurrent implementation of a decision support tool for identifying STEMI. Sex was assigned as indicated in the medical record. Data were analyzed in two groups: Pre-2016 (PRE) and 2016-2022 (POST). RESULTS: In total, 3,153 patients (1,301 PRE; 1,852 POST) were assessed; the average age was 65.2 years, 32.6% female, 87.7% white with significant differences in age and race between the two cohorts. Of the total 3,153 MI alerts, 239 were false activations, leaving 2,914 for analysis. 2,115 (72.6%) had cardiac catheterization while 16 (6.7%) of the 239 had a cardiac catheterization. There was an overall decrease in DTBT of 27.5% PRE to POST of prehospital ECG transmission (p < 0.001); PRE median time was 74.5 min vs. 55 min POST. There was no significant difference between rates of cardiac catheterization PRE and POST for all patients. After accounting for age, race, and mode of arrival, DTBT was 12.2% longer in women, as compared to men (p < 0.001) PRE vs. POST. DTBT among women was significantly shorter when comparing PRE to POST periods (median 77 min vs. 60 min; p = 0.0001). There was no significant sex difference in the proportion of those with cardiac catheterization between the two cohorts (62.5% vs. 63.5%; p = 0.73). CONCLUSION: Introduction of a decision support tool with prehospital ECG transmission with prehospital ECG transmission decreased overall DTBT by 20 min (27.5%). Women in the study had a 17-minute decrease in DTBT (22%), but their DTBT remained 12.2% longer than men for reasons that remain unclear.

Insight into the molecular basis of substrate recognition by the wall teichoic acid glycosyltransferase TagA.

Wall teichoic acid (WTA) polymers are covalently affixed to the Gram-positive bacterial cell wall and have important functions in cell elongation, cell morphology, biofilm formation, and ß-lactam antibiotic resistance. The first committed step in WTA biosynthesis is catalyzed by the TagA glycosyltransferase (also called TarA), a peripheral membrane protein that produces the conserved linkage unit, which joins WTA to the cell wall peptidoglycan. TagA contains a conserved GT26 core domain followed by a C-terminal polypeptide tail that is important for catalysis and membrane binding. Here, we report the crystal structure of the Thermoanaerobacter italicus TagA enzyme bound to UDP-N-acetyl-d-mannosamine, revealing the molecular basis of substrate binding. Native MS experiments support the model that only monomeric TagA is enzymatically active and that it is stabilized by membrane binding. Molecular dynamics simulations and enzyme activity measurements indicate that the C-terminal polypeptide tail facilitates catalysis by encapsulating the UDP-N-acetyl-d-mannosamine substrate, presenting three highly conserved arginine residues to the active site that are important for catalysis (R214, R221, and R224). From these data, we present a mechanistic model of catalysis that ascribes functions for these residues. This work could facilitate the development of new antimicrobial compounds that disrupt WTA biosynthesis in pathogenic bacteria.

Early production of proinflammatory cytokines in response to Scedosporium apiospermum during murine pulmonary infection.

Scedosporium apiospermum is an opportunistic pathogen that can cause pulmonary infections in both immunosuppressive and immunocompetent patients. Cytokines are molecules that mediate the immune response to promote or eliminate fungal infections. In this work, we evaluated the cytokines profile in the lung and serum of mice infected with Scedosporium apiospermum. We found early production of IL-6, IL-1ß and TNF-α cytokines in the lung of infected mice during the first 5 days of infection. We suggest that release of pro-inflammatory cytokines could play a role in the control of fungal invasion.

Safety profile of intravenous administration of live Pichia pastoris cells in mice.

Pichia pastoris has been widely used to produce antigenic proteins aimed to integrate subunit vaccines. Moreover, increasing interest in large-scale vaccine production at the lowest cost is rapidly focusing in the development of yeast surface display (YSD) systems for delivery of antigens. In this scenario, the safety of live yeast administration must be warranted, however, such information is very scarce. Here, we assess the intravenous administration (i.v.) of live P. pastoris cells in order to trace dissemination in BALB/c mice and to evaluate the immune response raised against the yeast compared to the well-defined pathogen Candida albicans. Our results demonstrate dissemination of P. pastoris to the heart, kidney, and spleen, but it is quickly eliminated during the first 48 h postinfection (hpi), with persistence in the liver along with mild mononuclear (MN) and polymorphonuclear (PMN) infiltrate, which was resolved at 144 hpi. In vivo delayed-type hypersensitivity test (DTH) or in vitro antigenic stimulation of mice splenocytes demonstrate that transient infection of P. pastoris did not induce a cell-mediated immune response nor increase the level of circulating IgG or IgM. These results demonstrate the innocuous profile of P. pastoris and support its use as a safe delivery system for vaccine development.

"House Calls" by Mobile Integrated Health Paramedics for Patients with Heart Failure: A Feasibility Study.

Background: Early readmissions following hospital discharge for heart failure (HF) remain a major concern. Among the various strategies designed to reduce readmissions, home evaluations have been observed to have a favorable impact. We assessed the feasibility of integrating community paramedics into the outpatient management of HF patients. Methods: Selected paramedics completed an educational HF curriculum. These Mobile Integrated Health Paramedics (MIHP) performed scheduled home visits 2- and 15-days post-discharge for patients with Stage C HF (Phase I) and patients with Stage D HF (Phase II). Facilitated by a Call Center, a process was created for performing urgent MIHP house calls within 60 minutes of a medical provider's request. A HF specialist, with an on-call emergency department command physician, could order an intravenous diuretic during home visits. During each phase of the study the incidence of 30-day HF readmissions, 30-day all-cause readmissions, emergency room evaluations, unplanned office encounters, as well as any adverse events were prospectively documented. Results: Collaborative relationships between our hospital network and local EMS organizations were created. There were 82 MIHP home visits. Eight patients received urgent home evaluations within 60-minutes post-request, one requiring transport to an ED. The incidence of all-cause 30-day readmissions in 20 Stage C and 20 Stage D patients was 15% and 40%, respectively. There were no adverse events attributable to the MIHP house calls. Conclusions: It is feasible to integrate MIHPs into the outpatient management of HF. Signals of effectiveness for reducing early readmissions were observed. Obstacles to creating an effective paramedic "House Calls" program were identified. A randomized trial is required to assess the value of this care process and its impact on early readmissions in patients with Stage C and Stage D HF.

Structure and mechanism of TagA, a novel membrane-associated glycosyltransferase that produces wall teichoic acids in pathogenic bacteria.

Staphylococcus aureus and other bacterial pathogens affix wall teichoic acids (WTAs) to their surface. These highly abundant anionic glycopolymers have critical functions in bacterial physiology and their susceptibility to ß-lactam antibiotics. The membrane-associated TagA glycosyltransferase (GT) catalyzes the first-committed step in WTA biosynthesis and is a founding member of the WecB/TagA/CpsF GT family, more than 6,000 enzymes that synthesize a range of extracellular polysaccharides through a poorly understood mechanism. Crystal structures of TagA from T. italicus in its apo- and UDP-bound states reveal a novel GT fold, and coupled with biochemical and cellular data define the mechanism of catalysis. We propose that enzyme activity is regulated by interactions with the bilayer, which trigger a structural change that facilitates proper active site formation and recognition of the enzyme's lipid-linked substrate. These findings inform upon the molecular basis of WecB/TagA/CpsF activity and could guide the development of new anti-microbial drugs.

Distinct innate immune responses between sublethal and lethal models of disseminated candidiasis in newborn BALB/c mice.

Invasive candidiasis is associated with a high incidence and mortality rates in infants, especially in preterm newborns. The immunopathogenesis of the mycosis during the neonatal period is poorly understood. Although several in vivo models exist to study invasive candidiasis, the majority of studies employ distinct routes of infection and use 2 to 6 day-old mice that could be less comparable in studying candidiasis in preterm infants. In this study, by using 0-days-old mice we developed a new neonatal murine model of intravenous Candida albicans infection. Using different inoculums of Candida albicans we evaluated survival, dissemination of the fungus, frequency of CD45+ cells, and cytokine production in the liver, brain, and kidneys of newborn and adult BALB/c mice. Unexpectedly, the newborn mice infected with a low inoculum (1×105 cfu per mouse) of Candida albicans survive to the infection. Compared to adult mice, the liver and brain of newborn animals had the greatest fungal burden, fungal invasion and leukocyte infiltrate. A moderate production of TNFα, IL-1ß, IL-6 and IFNγ was detected in tissues of newborn mice infected with a non-lethal inoculum of Candida albicans. In contrast, overproduction of TNFα, IL-1ß, IL-6 and IL-10 was determined when injecting with a lethal inoculum. In agreement, flow cytometry of brain and liver showed an inoculum-dependent CD45+ leukocyte infiltration in newborn mice infected with Candida albicans. Overall, our data shows that Candida albicans infection in newborn mice affects mainly the brain and liver and a 2-fold increase of the inoculum rapidly becomes lethal probably due to massive fungal invasion and exacerbated CD45+ leukocyte infiltrate and cytokine production. This study is the first analysis of innate immune responses in different tissues during early neonatal disseminated candidiasis.

SOP- habituation laboratory: An interactive tool for simulating the basic behavioral features of habituation.

This paper presents an open-source online tool for introducing psychology students to the major theoretical and empirical facts of habituation. The tool was designed in a way that combines theory and data through simulated experiments. The simulations exemplify how the priming theory of Allan R. Wagner accounts for the set of behavioral characteristics of habituation proposed by Richard F. Thompson and W. Alden Spencer in 1966. Through this interactive platform, the user can learn the basics of the theory and examine how it accounts for the empirical facts with different parameters. Instructions and commands are provided in three languages: English, Spanish, and Portuguese.

A model based on PDCA and data mining approach for the prevention of occupational accidents in the plumbing activity in the construction sector.

BACKGROUND: Occupational accidents in the plumbing activity in the construction sector in developing countries have high rates of work absenteeism. The productivity of enterprises is heavily influenced by it. OBJECTIVE: To propose a model based on the Plan, Do, Check, and Act cycle and data mining for the prevention of occupational accidents in the plumbing activity in the construction sector. METHODS: This cross-sectional study was administered on a total of 200 male technical workers in plumbing. It considers biological, biomechanical, chemical, and, physical risk factors. Three data mining algorithms were compared: Logistic Regression, Naive Bayes, and Decision Trees, classifying the occurrences occupational accident. The model was validated considering 20% of the data collected, maintaining the same proportion between accidents and non-accidents. The model was applied to data collected from the last 17 years of occupational accidents in the plumbing activity in a Colombian construction company. RESULTS: The results showed that, in 90.5% of the cases, the decision tree classifier (J48) correctly identified the possible cases of occupational accidents with the biological, chemical, and, biomechanical, risk factors training variables applied in the model. CONCLUSION: The results of this study are promising in that the model is efficient in predicting the occurrence of an occupational accident in the plumbing activity in the construction sector. For the accidents identified and the associated causes, a plan of measures to mitigate the risk of occupational accidents is proposed.

Blood glucose and epicardial adipose tissue at the hospital admission as possible predictors for COVID-19 severity.

PURPOSE: To study the possible association of CT-derived quantitative epicardial adipose tissue (EAT) and glycemia at the admission, with severe outcomes in patients with COVID-19. METHODS: Two hundred and twenty-nine patients consecutively hospitalized for COVID-19 from March 1st to June 30th 2020 were studied. Non contrast chest CT scans, to confirm diagnosis of pneumonia, were performed. EAT volume (cm3) and attenuation (Hounsfield units) were measured using a CT post-processing software. The primary outcome was acute respiratory distress syndrome (ARDS) or in-hospital death. RESULTS: The primary outcome occurred in 56.8% patients. Fasting blood glucose was significantly higher in the group ARDS/death than in the group with better prognosis [114 (98-144) vs. 101 (91-118) mg/dl, p = 0.001]. EAT volume was higher in patients with vs without the primary outcome [103 (69.25; 129.75) vs. 78.95 (50.7; 100.25) cm3, p < 0.001] and it was positively correlated with glycemia, PCR, fibrinogen, P/F ratio. In the multivariable logistic regression analysis, age and EAT volume were independently associated with ARDS/death. Glycemia and EAT attenuation would appear to be factors involved in ARDS/death with a trend of statistical significance. CONCLUSIONS: Our findings suggest that both blood glucose and EAT, easily measurable and modifiable targets, could be important predisposing factors for severe Covid-19 complications.

[Solid-pseudopapillary tumor of the pancreas. Presentation of 3 clinical cases]. / Tumor sólido-pseudopapilar del páncreas presentación de 3 casos clínicos.

The solid-pseudopapillary tumors of the pancreas (TSPP) are a uncommon neoplasm (1- 2% of exocrine tumors). Are more frequent in young women and presents a low malignant potential (15% develop metastases). Its preoperative diagnosis is very difficult, because these tumors haven`t radiological features that makes distinguish from the other pancreatic cysts tumors. Usually are a histopathological finding. Surgical therapy provides good survival, even in the presence of metastases. We present 3 cases with clinical, imaging, treatment and histopathology to provide more information about this rare disease.

The Sometimes Context-Specific Habituation: Theoretical Challenges to Associative Accounts.

A substantial corpus of experimental research indicates that in many species, long-term habituation appears to depend on context-stimulus associations. Some authors have recently emphasized that this type of outcome supports Wagner's priming theory, which affirms that responding is diminished when the eliciting stimulus is predicted by the context where the animal encountered that stimulus in the past. Although we agree with both the empirical reality of the phenomenon as well as the principled adequacy of the theory, we think that the available evidence is more provocative than conclusive and that there are a few nontrivial empirical and theoretical issues that need to be worked out by researchers in the future. In this paper, we comment on these issues within the framework of a quantitative version of priming theory, the SOP model.

In vitro characterization and preferential infection by canine distemper virus of glial precursors with Schwann cell characteristics from adult canine brain.

AIMS: Canine distemper virus (CDV)-induced demyelinating leukoencephalomyelitis is a naturally occurring model for multiple sclerosis. The aim of this study was to establish primary glial cell cultures from adult canine brain for the analysis of CDV spread and cell tropism. METHODS: Cultures were inoculated with the CDV-R252 and a CDV-Onderstepoort strain expressing the green fluorescent protein (CDV-OndeGFP). CDV antigen expression was studied using cell type-specific antibodies at different days post infection. Glial cells expressing p75(NTR) were purified using antibody-based techniques and characterized with regard to antigen expression and proliferation. RESULTS: Three weeks after seeding, cultures contained spindle-shaped cells expressing p75(NTR), oligodendrocytic cells, astrocytes, microglia and fibroblasts. Both CDV strains induced a mild to moderate cytopathic effect that consisted of single necrotic and few syncytial giant cells, but displayed in part a differential cell tropism. Whereas CDV-OndeGFP expression in microglia and astrocytes did not exceed 1% and 50%, respectively, CDV-R252 infected 100% and 80% of both cell types, respectively. The cells most early infected by both CDV strains expressed p75(NTR) and may correlate to cells previously identified as aldynoglia. Treatment of p75(NTR+) cells with Schwann cell mitogens and serum deprivation increased proliferation and A2B5 expression, respectively, indicating common properties compared with Schwann cells and oligodendrocyte precursors. CONCLUSIONS: Infection of adult canine astrocytes and microglia revealed CDV strain-specific cell tropism. Moreover, this is the first identification of a glial cell type with Schwann cell-like properties in adult canine brain and, more importantly, these cells displayed a high susceptibility to CDV infection.

Solution of Heliospheric Propagation: Unveiling the Local Interstellar Spectra of Cosmic-ray Species.

Local interstellar spectra (LIS) for protons, helium, and antiprotons are built using the most recent experimental results combined with state-of-the-art models for propagation in the Galaxy and heliosphere. Two propagation packages, GALPROP and HelMod, are combined to provide a single framework that is run to reproduce direct measurements of cosmic-ray (CR) species at different modulation levels and at both polarities of the solar magnetic field. To do so in a self-consistent way, an iterative procedure was developed, where the GALPROP LIS output is fed into HelMod, providing modulated spectra for specific time periods of selected experiments to compare with the data; the HelMod parameter optimization is performed at this stage and looped back to adjust the LIS using the new GALPROP run. The parameters were tuned with the maximum likelihood procedure using an extensive data set of proton spectra from 1997 to 2015. The proposed LIS accommodate both the low-energy interstellar CR spectra measured by Voyager 1 and the high-energy observations by BESS, Pamela, AMS-01, and AMS-02 made from the balloons and near-Earth payloads; it also accounts for Ulysses counting rate features measured out of the ecliptic plane. The found solution is in a good agreement with proton, helium, and antiproton data by AMS-02, BESS, and PAMELA in the whole energy range.

Characterization of a recurrent germ line mutation of the E-cadherin gene: implications for genetic testing and clinical management.

PURPOSE: To identify germ line CDH1 mutations in hereditary diffuse gastric cancer (HDGC) families and develop guidelines for management of at risk individuals. EXPERIMENTAL DESIGN: We ascertained 31 HDGC previously unreported families, including 10 isolated early-onset diffuse gastric cancer (DGC) cases. Screening for CDH1 germ line mutations was done by denaturing high-performance liquid chromatography and automated DNA sequencing. RESULTS: We identified eight inactivating and one missense CDH1 germ line mutation. The missense mutation conferred in vitro loss of protein function. Two families had the previously described 1003C>T nonsense mutation. Haplotype analysis revealed this to be a recurrent and not a founder mutation. Thirty-six percent (5 of 14) of the families with a documented DGC diagnosed before the age of 50 and other cases of gastric cancer carried CDH1 germ line mutations. Two of 10 isolated cases of DGC in individuals ages <35 years harbored CDH1 germ line mutations. One mutation positive family was ascertained through a family history of lobular breast cancer (LBC) and another through an individual with both DGC and LBC. Occult DGC was identified in five of six prophylactic gastrectomies done on asymptomatic, endoscopically negative 1003C>T mutation carriers. CONCLUSIONS: In addition to families with a strong history of early-onset DGC, CDH1 mutation screening should be offered to isolated cases of DGC in individuals ages <35 years and for families with multiple cases of LBC, with any history of DGC or unspecified GI malignancies. Prophylactic gastrectomy is potentially a lifesaving procedure and clinical breast screening is recommended for asymptomatic mutation carriers.

GALACTIC COSMIC RAYS IN THE LOCAL INTERSTELLAR MEDIUM: VOYAGER 1 OBSERVATIONS AND MODEL RESULTS.

Since 2012 August Voyager 1 has been observing the local interstellar energy spectra of Galactic cosmic-ray nuclei down to 3 MeV nuc-1 and electrons down to 2.7 MeV. The H and He spectra have the same energy dependence between 3 and 346 MeV nuc-1, with a broad maximum in the 10-50 MeV nuc-1 range and a H/He ratio of 12.2 ± 0.9. The peak H intensity is ~15 times that observed at 1 AU, and the observed local interstellar gradient of 3-346 MeV H is -0.009 ± 0.055% AU-1, consistent with models having no local interstellar gradient. The energy spectrum of electrons (e - + e +) with 2.7-74 MeV is consistent with E -1.30±0.05 and exceeds the H intensity at energies below ~50 MeV. Propagation model fits to the observed spectra indicate that the energy density of cosmic-ray nuclei with >3 MeV nuc-1 and electrons with >3 MeV is 0.83-1.02 eV cm-3 and the ionization rate of atomic H is in the range of 1.51-1.64 × 10-17 s-1. This rate is a factor >10 lower than the ionization rate in diffuse interstellar clouds, suggesting significant spatial inhomogeneity in low-energy cosmic rays or the presence of a suprathermal tail on the energy spectrum at much lower energies. The propagation model fits also provide improved estimates of the elemental abundances in the source of Galactic cosmic rays.

BAYESIAN ANALYSIS OF COSMIC RAY PROPAGATION: EVIDENCE AGAINST HOMOGENEOUS DIFFUSION.

We present the results of the most complete scan of the parameter space for cosmic ray (CR) injection and propagation. We perform a Bayesian search of the main GALPROP parameters, using the MultiNest nested sampling algorithm, augmented by the BAMBI neural network machine-learning package. This is the first study to separate out low-mass isotopes (p, p ¯ , and He) from the usual light elements (Be, B, C, N, and O). We find that the propagation parameters that best-fit p, p ¯ , and He data are significantly different from those that fit light elements, including the B/C and 10Be/9Be secondary-to-primary ratios normally used to calibrate propagation parameters. This suggests that each set of species is probing a very different interstellar medium, and that the standard approach of calibrating propagation parameters using B/C can lead to incorrect results. We present posterior distributions and best-fit parameters for propagation of both sets of nuclei, as well as for the injection abundances of elements from H to Si. The input GALDEF files with these new parameters will be included in an upcoming public GALPROP update.