Low-level Response Research Articles

Superiority in Rhesus Macaques of Targeting HIV-1 Env gp140 to CD40 versus LOX-1 in Combination with Replication-Competent NYVAC-KC for Induction of Env-Specific Antibody and T Cell Responses.

We compared the HIV-1-specific immune responses generated by targeting HIV-1 envelope protein (Env gp140) to either CD40 or LOX-1, two endocytic receptors on dendritic cells (DCs), in rhesus macaques primed with a poxvirus vector (NYVAC-KC) expressing Env gp140. The DC-targeting vaccines, humanized recombinant monoclonal antibodies fused to Env gp140, were administered as a boost with poly-ICLC adjuvant either alone or coadministered with the NYVAC-KC vector. All the DC-targeting vaccine administrations with poly-ICLC increased the low-level serum anti-Env IgG responses elicited by NYVAC-KC priming significantly more (up to a P value of 0.01) than in a group without poly-ICLC. The responses were robust and cross-reactive and contained antibodies specific to multiple epitopes within gp140, including the C1, C2, V1, V2, and V3, C4, C5, and gp41 immunodominant regions. The DC-targeting vaccines also elicited modest serum Env-specific IgA responses. All groups gave serum neutralization activity limited to tier 1 viruses and antibody-dependent cytotoxicity responses (ADCC) after DC-targeting boosts. Furthermore, CD4+ and CD8+ T cell responses specific to multiple Env epitopes were strongly boosted by the DC-targeting vaccines plus poly-ICLC. Together, these results indicate that prime-boost immunization via NYVAC-KC and either anti-CD40.Env gp140/poly-ICLC or anti-LOX-1.Env gp140/poly-ICLC induced balanced antibody and T cell responses against HIV-1 Env. Coadministration of NYVAC-KC with the DC-targeting vaccines increased T cell responses but had minimal effects on antibody responses except for suppressing serum IgA responses. Overall, targeting Env to CD40 gave more robust T cell and serum antibody responses with broader epitope representation and greater durability than with LOX-1.IMPORTANCE An effective vaccine to prevent HIV-1 infection does not yet exist. An approach to elicit strong protective antibody development is to direct virus protein antigens specifically to dendritic cells, which are now known to be the key cell type for controlling immunity. In this study, we have tested in nonhuman primates two prototype vaccines engineered to direct the HIV-1 coat protein Env to dendritic cells. These vaccines bind to either CD40 or LOX-1, two dendritic cell surface receptors with different functions and tissue distributions. We tested the vaccines described above in combination with attenuated virus vectors that express Env. Both vaccines, but especially that delivered via CD40, raised robust immunity against HIV-1 as measured by monitoring potentially protective antibody and T cell responses in the blood. The safety and efficacy of the CD40-targeted vaccine justify further development for future human clinical trials.

Supervised classification of etoposide-treated in vitro adherent cells based on noninvasive imaging morphology.

Single-cell studies using noninvasive imaging is a challenging, yet appealing way to study cellular characteristics over extended periods of time, for instance to follow cell interactions and the behavior of different cell types within the same sample. In some cases, e.g., transplantation culturing, real-time cellular monitoring, stem cell studies, in vivo studies, and embryo growth studies, it is also crucial to keep the sample intact and invasive imaging using fluorophores or dyes is not an option. Computerized methods are needed to improve throughput of image-based analysis and for use with noninvasive microscopy such methods are poorly developed. By combining a set of well-documented image analysis and classification tools with noninvasive microscopy, we demonstrate the ability for long-term image-based analysis of morphological changes in single cells as induced by a toxin, and show how these changes can be used to indicate changes in biological function. In this study, adherent cell cultures of DU-145 treated with low-concentration (LC) etoposide were imaged during 3 days. Single cells were identified by image segmentation and subsequently classified on image features, extracted for each cell. In parallel with image analysis, an MTS assay was performed to allow comparison between metabolic activity and morphological changes after long-term low-level drug response. Results show a decrease in proliferation rate for LC etoposide, accompanied by changes in cell morphology, primarily leading to an increase in cell area and textural changes. It is shown that changes detected by image analysis are already visible on day 1 for [Formula: see text] etoposide, whereas effects on MTS and viability are detected only on day 3 for [Formula: see text] etoposide concentration, leading to the conclusion that the morphological changes observed occur before and at lower concentrations than a reduction in cell metabolic activity or viability. Three classifiers are compared and we report a best case sensitivity of 88% and specificity of 94% for classification of cells as treated/untreated.

Antibiotics promote the sampling of luminal antigens and bacteria via colonic goblet cell associated antigen passages

ABSTRACTBacterial translocation is defined as the passage of live bacteria from the gut lumen to distant sites. Gut commensal bacteria translocation has been attributed to ‘leakiness’, or ‘barrier breach’ of the intestinal epithelium, allowing live bacteria to cross an inappropriately permeable barrier and disseminate to distant sites. Alternatively, studies suggest dendritic cells directly capture luminal commensal bacteria and transport them to distant sites in the steady-state by extending dendrites between epithelial cells into the lumen. Recently we identified translocation of commensal gut bacteria following antibiotics was associated with the formation of goblet cell associated antigen passages (GAPs) in the colon and dependent upon goblet cells (GCs). The translocation of native gut commensal bacteria resulted in low-level inflammatory responses and potentiated mucosal damage in response to concurrent epithelial injury. Here we extend these observations and demonstrate properties of colonic GAPs and observations supporting their priority in the translocation of colonic commensal bacteria.

Design Considerations for Robust Noise Rejection in Otoacoustic Emissions Measured In-Field Using Adaptive Filtering

Otoacoustic emission (OAE) measurement is a sensitive and effective technique to monitor changes in the inner ear potentially induced by noise exposure. However, outside a controlled testing environment, measurements are challenging since the level of ambient noise might be higher than the low-level OAE response. Therefore, an OAE system was designed, suitable to measure OAEs repeatedly on an individual worker in noisy test conditions. This system features a left and right earpiece, each equipped with a pair of miniature loudspeakers, an external and an internal microphone. In addition to the passive attenuation of the earpiece, adaptive filtering on the distortion product OAE (DPOAE) signals is used to further enhance the ratio between the measured OAE signal and interfering noise. The adaptive filtering technique uses the sound captured by the ipsilateral external and internal microphones as well as from the contralateral internal and external microphones. In this paper, the accuracy of DPOAE signals are studied using different combinations of the four microphones in single and dual adaptive filter topologies, as well as the benefits of adding a fixed transfer function in the adaptive filtering algorithm topology to estimate the acoustic path. Side-by-side comparison shows that a dual-stage adaptive filtering algorithm, using a combination of the contralateral internal microphone with the ipsilateral external microphone, is the most promising approach to denoise the DPOAE signal.

Abstract ES4-3: Tumor-infiltrating lymphocytes (TILs) and genomic signatures of immune cells

Abstract Increased levels of tumor-infiltrating lymphocytes have been described in subsets of breast cancer, in particular triple-negative (TNBC) and HER2-positive (HER2+) tumors. Neoadjuvant therapy approaches in breast cancer have been used as a strategy to characterize subgroups of tumors with increased tumor-infiltrating lymphocytes (TILs). We have described tumor-infiltrating lymphocytes (TILs) as predictors of pathological complete response to neoadjuvant chemotherapy in breast cancer trials conducted by the German Breast Group. It has been shown that increased TILs are linked to an increased response rate to neoadjuvant therapy and improved prognosis after adjuvant therapy, in particular in triple-negative and HER2 positive breast cancer. In addition to tumor-infiltrating lymphocytes, immune mRNA markers have been studied in breast cancer and are associated with increased chemotherapy response and improved prognosis. These results suggest that some subtypes of breast cancer are immunogenic and that there is an ongoing low-level immune response present even in clinically progressing tumors. Therapeutic approaches to modulate tumor-associated inflammation by immune checkpoint inhibitors have shown clinical responses in a subset of triple-negative breast cancers, and some of these responses were durable. These targeted therapies have stimulated interest in the interaction between immune cells and tumor cells. Based on the concept that conventional chemotherapy might be immunogenic, some studies are evaluating the combination of an immune checkpoint inhibitor and a chemotherapy. Considering the fact that responses to immune checkpoint inhibitors are observed in only a subset of tumors, biomarkers that will be able to identify those tumors are needed. Possible biomarkers include the therapeutic targets, in particular PD-L1 and PD1, tumor-infiltrating lymphocytes, predictive mRNA signatures, immunogenic neoepitope signatures that are generated by mutations as well as other factors. Due to the low costs of a simple H&E based test, tumor infiltrating lymphocytes might become a promising biomarker as part of the pathology report. In preparation for this, the international immune-oncology working group (TIL working group) has published standardized guidelines for TIL evaluation in breast cancer, and has conducted a first international TIL ring trial. This ring trial suggests that decentral TIL evaluation is feasible. In some situations, in particular in tumors with intermediate and heterogeneous TILs, additional tests, including gene expression signatures might be necessary. It should be emphasized that the clinical utility of TILs for current medical decisions is very limited at the present time, and the ultimate usefulness of these biomarker concepts will be obtained in the ongoing and future clinical immunotherapy trials. The presentation will provide an overview on recent results on immune markers for prediction of response to neoadjuvant chemotherapy and prognosis with a focus in standardized histomorphological evaluation of tumor-infiltrating lymphocytes as well as clinical trials concepts and biomarker strategies for future immune checkpoint inhibitor therapies in breast cancer. Citation Format: Denkert C. Tumor-infiltrating lymphocytes (TILs) and genomic signatures of immune cells [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr ES4-3.

HIV-1 Env DNA prime plus gp120 and gp70-V1V2 boosts induce high level of V1V2-specific IgG and ADCC responses and low level of Env-specific IgA response: implication for improving RV144 vaccine regimen

Emerging Microbes & Infections (2017) 6, e102; doi:10.1038/emi.2017.90; published online 29 November 2017

Prospects for improving neovascularization of the ischemic heart: Lessons from development.

Neovascularization of the ischemic myocardium postinfarction is necessary to restore blood flow to vulnerable cardiomyocytes and will be indispensable for prospective regenerative strategies, to perfuse newly formed myocardium. Therapeutic attempts to enhance new vessel formation have, to date, yielded modest clinical benefits, and innovative approaches are now needed. Intrinsic mechanisms are initiated by the heart in an attempt to rebuild injured vessels, but these are poorly understood. Insight into the underlying mechanisms may reveal targets for therapeutically augmenting this low-level neovascular response. Starting from a limited number of descriptive studies, this review summarizes what is known of coronary neovascularization and explores putative mechanisms and cellular sources which may endogenously contribute, or that may be pharmacologically triggered, to support vasculo- or angiogenesis. As injury responses in the adult frequently recapitulate embryological processes, a particular focus is placed on the developmental mechanisms of coronary vessel formation. An understanding of the cellular sources and the regulatory pathways used by the embryo may reveal novel targets for reactivating coronary vessel and myocardial regeneration.

Highly Neurovirulent Viruses Invade and Spread in the Brain Using the Host Reactions Triggered by Infection

Coronaviruses (CoVs) have relatively high mutation and RNA recombination rates and undergo rapid crossspecies transmission events in vitro and in vivo. The viruses have evolved diverse strategies to evade, blocking host immune responses, and resulting in an emergence of highly virulent CoVs to cause fatal diseases in human, such as severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). Among CoVs used for experimental studies, the cl-2 virus (cl-2), isolated from a neuropathogenic JHM viral strain (JHMV) of mouse hepatitis virus (MHV), shows extremely high virulence. All mice infected with cl-2 die within 3 days post-inoculation (dpi). Less virulent but still highly neurovirulent mutant viruses, soluble receptor resistant mutant virus (srr7) and Mu-3 virus (Mu-3), have been isolated from cl-2 and srr7, respectively. Although these viruses cause different pictures of neuropathologies with different distribution of viral antigens after infection, they use the same pathway to enter the brain before inducing lesions in the brain parenchyma. The pathway is formed as an astrocytic reticular network (ARN) after infection. The ARN functions as a conduit system in a similar way to a fibroblastic reticular network (FRN) in lymphoid organs. In addition, the viruses induce an immunosuppressive state in infected mice, mimicking endotoxin tolerance (ET), or lipopolysaccharide (LPS) tolerance. The immunosuppressive states are detected by; 1) pathological findings as 1a); a very low -level inflammatory response, 1b); the shrunken spleen, 1c); the refractory state to LPS challenge, and 1d); the local production of anti-inflammatory cytokines by neurons, and by 2) immunological studies, which showed 2a); the systemic production of anti-inflammatory cytokines by eliciting CD11b+Gra-1+ suppressor monocytes/macrophages (Mo/Mas).In addition, cultured splenic cells and Mo/Mas after infection exhibit 2b); low-level responses to stimulation of toll-like receptors (TLRs) 4 and 7, and 2c); the induction of Lewis X carbohydrate structure (Lex), which can trigger the C-type lectin-like receptor that initiates an immunosuppressive pathway, respectively.

Childhood trauma and mixed episodes are associated with poor response to lithium in bipolar disorders.

Reliable predictors of response to lithium are still lacking in bipolar disorders (BDs). However, childhood trauma has been hypothesized to be associated with poor response to lithium. We included 148 patients with BD, euthymic when retrospectively and clinically assessed for response to lithium and childhood trauma using reliable scales. According to the 'Alda scale', the sample consisted in 20.3% of excellent responders, 49.3% of partial responders and 30.4% of non-responders to lithium. A higher level of physical abuse significantly correlated with a lower level of response to lithium (P = 0.009). As compared to patients not exposed to any abuse, patients with at least two trauma abuses (emotional, physical or sexual) were more at risk of belonging to the non-responders group (OR = 4.91 95% CI (1.01-27.02)). Among investigated clinical variables, lifetime presence of mixed episodes and alcohol misuse were associated with non-response to lithium. Multivariate analyses demonstrated that physical abuse and mixed episodes were independently associated with poor response to lithium (P = 0.005 and P = 0.013 respectively). Childhood physical abuse might be involved in a poor future response to lithium prophylaxis, this effect being independent of the association between clinical expression of BD and poor response to lithium.

Evaluation of Some Immunostimulants on the Immune-response of Broiler Chickens Against Avian Influenza and Newcastle Diseases Vaccination

Immunostimulants get an increasing importance to enhance the immune system and allowing maximum performance in poultry production. The effects of using different immunostimulants on the immune response of broiler chickens to Newcastle disease (ND) and Avian influenza (AI) vaccination were investigated in this study. 100 day-old Ross broiler chicks were randomly divided into 4 groups, 25 birds for each group. Group A was not treated with any immunostimulants and used as a control, while groups B, C and D were supplemented with Imutrix® (oregano oils plus β-glucan), Evit liquid® (Vitamin E plus selenium) and inmunair®17.5 (Propionibacterium acnes and E. coli lipopolysaccharides), respectively. Statistical analysis results demonstrated significant divergence (P < 0.05) in body weight (BW) and feed conversion ratio among the applicable treatments. Birds of group B had the heaviest body weight (2.03 kg), followed by group C (1.87 Kg) and D (1.82 Kg) in comparison to the birds of group A (control, 1.79 Kg). Bursa of Fabricius weight showed non-significant differences among groups except in group B that showed an increase in the Bursa of Fabricius weight more than other groups especially the control group (1.15 g more). Comparable immune response was recorded for both AI and ND vaccination. The highest mean antibody titers were recorded for birds in group B at 14, 28 and 35 days old, while those in group D had the lowest antibody titre values. Thus, supplementing immunostimulants especially Imutrix® and Evit liquid® had a significant positive effect on performance characteristics and immune response against AI and ND vaccination in broiler chickens in contrast to non-treated group, which had low levels of immune response, rapid decreasing and no persistence for keeping high HI titer for long time.

Potential role of cyanidin 3-glucoside (C3G) in diabetic cardiomyopathy in diabetic rats: An in vivo approach.

The present study aimed to evaluate the importance of cyanidin 3-glucoside (C3G) of diabetic cardiomyopathy in diabetic rats. The rats were induced with diabetic using streptozotocin and total triglyceride (TG) and total cholesterol (TC) were determined. The range of myocardial enzymes such as aspartate aminotransferase (AST), creatine kinase (CK) and lactate dehydrogenase (LD) were also estimated, further, the Immuno histochemical analysis and western blot investigation were determined for the actual activity of C3G. Results indicated that the marker enzymes such as CK, LD and AST were significantly (P < 0.05) increased in STZ administered rats (DM group), while the levels of these elevated marker enzymes of cardiac injury significantly (P < 0.05) declined in the DM + C3G group, as compared to the diabetic group of rats. Additionally, a decrease in the level of TNF-alpha and interleukin-6, was noticed in the C3G treated group as compared to diabetic group. Finally, blotting analysis clearly confirmed that theC3G treatment resulted to higher level response of Bcl-2 and lower level response of caspase-3 and BAX. In conclusion, C3G a natural antioxidant may prevent cardiovascular complications by ameliorating oxidative damage, inflammation, metabolic dysfunctions and apoptosis pathways in type 2 diabetes.

Influence of HLA-DRB1 Alleles on the Variations of Antibody Response to Tuberculosis Serodiagnostic Antigens in Active Tuberculosis Patients.

Serology-based tests for tuberculosis (TB) diagnosis, though rapid, efficient and easily implemented, have so far shown unsatisfactory levels of sensitivity and specificity, probably due to variations of the antibody response in TB patients. The number and types of seropositive antigens vary from individual to individual. The person-to-person variations of antigen recognition may be linked to genetic polymorphisms of the human leukocyte antigen (HLA) class II alleles. In the present study, we find that there is a significant increase in the frequency of HLA-DRB1*14 (P = 2.5×10−4) among subjects with high antibody response levels compared to those with low antibody levels. HLA-DRB1*15, the most frequent allelic group in the studied active TB population, positively correlates with subjects with low antibody response levels rather than subjects with high antibody response levels (P = 0.005), which indicates the loss of relevant antigens for screening of patients with this allelic group. The potential association between HLA-DRB1 allelic group and individual antigens implies that TB diagnostic yield could be improved by the addition of antigens screened at the proteome scale in infected subjects from the HLA-DRB1*15 allelic group.

Plastic changes along auditory pathway during salicylate-induced ototoxicity: Hyperactivity and CF shifts

High dose of salicylate, the active ingredient in aspirin, has long been known to induce transient hearing loss, tinnitus and hyperacusis making it a powerful experimental tool. These salicylate-induced perceptual disturbances are associated with a massive reduction in the neural output of the cochlea. Paradoxically, the diminished neural output of the cochlea is accompanied by a dramatic increase in sound-evoked activity in the auditory cortex (AC) and several other parts of the central nervous system. Exactly where the increase in neural activity begins and builds up along the central auditory pathway are not fully understood. To address this issue, we measured sound-evoked neural activity in the cochlea, cochlear nucleus (CN), inferior colliculus (IC), and AC before and after administering a high dose of sodium salicylate (SS, 300 mg/kg). The SS-treatment abolished low-level sound-evoked responses along the auditory pathway resulting in a 20–30 dB threshold shift. While the neural output of the cochlea was substantially reduced at high intensities, the neural responses in the CN were only slightly reduced; those in the IC were nearly normal or slightly enhanced while those in the AC considerably enhanced, indicative of a progress increase in central gain. The SS-induced increase in central response in the IC and AC was frequency-dependent with the greatest increase occurring in the mid-frequency range the putative pitch of SS-induced tinnitus. This frequency-dependent hyperactivity appeared to result from shifts in the frequency receptive fields (FRF) such that the response areas of many FRF shifted/expanded toward the mid-frequencies. Our results suggest that the SS-induced threshold shift originates in the cochlea. In contrast, enhanced central gain is not localized to one region, but progressively builds up at successively higher stage of the auditory pathway either through a loss of inhibition and/or increased excitation.

Alcohol-related blackouts across 55 weeks of college: Effects of European-American ethnicity, female sex, and low level of response to alcohol

BackgroundWhile high blood alcohol concentrations (BACs) are required for alcohol-related blackouts (ARBs), additional characteristics also contribute to the risk, including a person’s ethnicity, sex, and phenotypes relating to heavier drinking. Few prospective studies of ARBs have evaluated how these additional characteristics interact. MethodData regarding 398 European American (EA), Asian and Hispanic students were extracted from a 55-week prospective study of different approaches to decrease heavy drinking among college freshmen. Information on past month ARB frequency was determined at 8 assessments. While controlling for the prior month maximum BAC and active education vs. control group assignment, the patterns and intensities of ARBs over time across ethnic groups were evaluated with ANOVA at each follow-up for the full sample, and then separately by sex and then by low vs. high levels of response to alcohol status (LR). The overall pattern of ARBs over time was evaluated with a 3 ethnic groups by 2 sexes by 2 LR status by 8 time points mixed-design ANOVA. ResultsHigher rates of ARBs over time were associated with EA ethnicity, female sex and a low LR to alcohol, with the ethnic differences in ARBs most robust in females and drinkers with high LRs. Participation in education programs aimed at heavy drinking was associated with decreases in ARBs. ConclusionsThe data indicate that in addition to BACs achieved, propensities toward ARBs relate to complex interactions between additional risk factors, including ethnicity, sex, and LR status.

Cell migration, viability and tissue reaction of calcium hypochlorite based-solutions irrigants: An in vitro and in vivo study

ObjectiveThis study aimed to analyze in vitro cytotoxicity to cultured 3T3 fibroblasts and in vivo inflammatory reaction in rats by calcium hypochlorite (Ca(OCl)2) solutions compared with sodium hypochlorite (NaOCl) solutions. DesignCultured 3T3 fibroblasts were exposed to different concentrations of (Ca(OCl)2) and NaOCl solutions, and a scratch assay was performed. The viability rate was analyzed with trypan blue assay. Both solutions of 1% and 2.5% concentrations were injected into the subcutaneous tissue of 18 male Wistar rats aged 18 weeks. The inflammatory tissue reaction was evaluated at 2h, 24h, and 14days after the injections. The samples were qualitatively analyzed using a light microscope. Statistical analysis was performed with ANOVA and Tukey post hoc tests for in vitro assays and Kruskal–Wallis and Dunn post hoc tests for in vivo assays (α=0.05). ResultsIn the scratch assay, Ca(OCl)2 showed no significant difference compared with the control group (culture medium) at 24h (p<0.05). Solutions of 0.0075% and 0.005% NaOCl and Ca(OCl)2 concentrations presented similar results compared with those in the positive control group (hydrogen peroxide) (p>0.05) in the trypan blue assay. In the in vivo assay, 1% Ca(OCl)2 group showed a significant decrease in neutrophils at 2h and 24h (p=0.041) and 2h and 14days (p=0.017). There was no statistically significant difference for lymphocyte/plasmocyte and macrophage counts among the different concentration groups. ConclusionsCa(OCl)2 showed favorable results of viability and induced a low-level inflammatory response. Ca(OCl)2 presented acceptable cytotoxicity and biocompatibility as an irrigant solution.

Changes in some pro-and anti-inflammatory cytokines produced by bovine peripheral blood mononuclear cells following foot and mouth disease vaccination

Interleukin (IL)-17 is exclusively produced by CD4 helper T-cells upon activation. It most often acts as a pro-inflammatory cytokine, which stimulates the release of pro-inflammatory cytokines IL-6, IL-8, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF). In this study, we studied the in-vitro IL-17 response to specific antigens and a variety of mitogens and compared the IL-17 response to IL-2, IL-4, IL-5, IL-6, IL-10, and IFN-γ responses. We used a foot and mouth disease (FMD) vaccine as specific antigens and mitogens (phytohemagglutinin [PHA], pokeweed mitogen [PWM], and concanavalin A [Con A]) to stimulate peripheral blood mononuclear cells (PBMCs) of vaccinated calves. Cell culture supernatant was harvested and analyzed for cytokines, using commercially available bovine ELISA kits. The mitogens induced a significant increase in IL-17 production. IL-17 was produced at high levels in response to the T cell-stimulated mitogens, PHA, and Con A, and at low levels in response to PWM mitogens. In contrast, level of the produced IL-17 cytokines in response to the FMDV antigens was lower as compared to those produced by mitogens. The FMDV antigens and mitogens significantly increased IL-17 production. There was not a correlation between IL-17 production and type-1 cytokine, IFN-γ, and IL-2, while there was a correlation between type-2 cytokine, IL-4, and IL-5 at either cytokine level produced by PBMCs stimulated by FMDV antigens. Moreover, there was an interaction between IL-17 and IL-6, that is, as IL-6 cytokine level elevated or diminished, IL-17 cytokine level increased or decreased, as well.

Measuring progress in maternal and newborn health care in Mexico: validating indicators of health system contact and quality of care.

BackgroundThe majority of births in Mexico take place in a health facility and are attended by a skilled birth attendant, yet maternal mortality has not declined to anticipated levels. Coverage estimates of skilled attendance and other maternal and newborn interventions often rely on women’s self-report through a population-based survey, the accuracy of which is not well established.MethodsWe used a facility-based design to validate women’s report of skilled birth attendance, as well as other key elements of maternal, newborn intrapartum, and immediate postnatal care. Women’s reports of labor and delivery care were collected by exit interview prior to hospital discharge and were compared against direct observation by a trained third party in a Mexican public hospital (n = 597). For each indicator, validity was assessed at the individual level using the area under the receiver operating curve (AUC) and at the population level using the inflation factor (IF).ResultsFive of 47 indicators met both validation criteria (AUC > 0.60 and 0.75 < IF < 1.25): urine sample screen, injection or IV medication received during labor, before the birth of the baby (i.e., uterotonic for either induction or augmentation of labor), episiotomy, excessive bleeding, and receipt of blood products. An additional 9 indicators met criteria for the AUC and 18 met criteria for the IF. A skilled attendant indicator had high sensitivity (90.1 %: 95 % CI: 87.1–92.5 %), low specificity (14.0 %: 95 % CI: 5.8–26.7 %) and was suitable for population-level estimation only.ConclusionWomen are able to give valid reports on some aspects of the content of care, although questions regarding the indication for interventions are less likely to be known. Questions that include technical terms or refer to specific time periods tended to have lower response levels. A key aspect of efforts to improve maternal and newborn health requires valid measurement of women’s access to maternal and newborn health interventions and the quality of such services. Additional work on improving measurement of population coverage indicators is warranted.Electronic supplementary materialThe online version of this article (doi:10.1186/s12884-016-1047-0) contains supplementary material, which is available to authorized users.

Robust adaptive detection of buried pipes using GPR

Detection of buried objects such as pipes using a Ground Penetrating Radar (GPR) is intricate for three main reasons. First, noise is important in the resulting image because of the presence of several rocks and/or layers in the ground, highly influencing the Probability of False Alarm (PFA) level. Also, wave speed and object responses are unknown in the ground and depend on the relative permittivity, which is not directly measurable. Finally, the depth of the pipes leads to strong attenuation of the echoed signal, leading to poor SNR scenarios. In this paper, we propose a detection method: (1) enhancing the signal of interest while reducing the noise and layer contributions, and (2) giving a local estimate of the relative permittivity. We derive an adaptive detector where the signal of interest is parametrised by the wave speed in the ground. For this detector, noise is assumed to follow a Spherically Invariant Random Vector (SIRV) distribution in order to obtain a robust detection. We use robust maximum likelihood-type covariance matrix estimators called M-estimators. To handle the significant amount of data, we consider regularised versions of said estimators. Simulation will allow to estimate the relation PFA-Threshold. Comparison is performed with standard GPR processing methods, showing the aptitude of the method in detecting pipes having low response levels with a reasonable PFA.

Траснформация обратной связм при осуществлении маркетинговых коммуникаций в интернет-пространстве

The article discusses the change process of feedback received from customers, modern companies originating in the digital environment. It was analyzed the hierarchy of possible options of consumer reaction to information received from the company.&#x0D; The author identifies the reasons for the low level of consumer response&#x0D; to received marketing information. A typology of consumer response to the communicative impact the company was offered.&#x0D; The type of negative consumer reactions, as well as the reasons for&#x0D; which the consumer generally does not communicate its opinion&#x0D; to the company was analyzed. The tools by which feedback can be&#x0D; arranged on the Internet, especially the interaction with consumers&#x0D; was described. The main characteristics that distinguish communication&#x0D; support of the company were identified. The issue of&#x0D; evaluation and performance review feedback due to the integrated&#x0D; use of marketing communication tools was designated

430. Antibodies to Recombinant Human α-L-Iduronidase Enhance Uptake into Macrophages in Murine Mucopolysaccharidosis Type I

Mucopolysaccharidosis I (MPS I) is an inherited, and progressive lysosomal storage disease caused by lack or low level of α-L-iduronidase (IDU), altering the catabolism of glycosaminoglycans (GAG) in lysosomes. Some clinical improvement in patients has been observed by enzyme replacement therapy with recombinant human IDU (rhIDU), but studies suggested that anti-rhIDU antibodies reduce the effectiveness of treatment. Most MPS I treated patients develop a humoral immune response against rhIDU. To determine how anti-rhIDU antibodies alter the tissue and cellular distribution of rhIDU in vivo, we sensitized MPS I mice to rhIDU by administering rhIDU via tail-vein injection from 4 to 16 weeks of age. Mice that developed anti-rhIDU antibodies showed a shift in rhIDU uptake preferentially towards tissues with high reticuloendothelial (RE) content (liver, spleen, thymus) vs. low RE content (lung, kidney, heart, brain). Furthermore, in mice with anti-rhIDU antibodies, rhIDU was mainly localized in Kupffer cells, with relatively less available for hepatocytes. In order to further understand how the humoral immune response against rhIDU alters its distribution, we comparedin vitro uptake of rhIDU in the absence or presence of antibodies against rhIDU in MPS I fibroblasts and macrophages. Bone marrow monocytes were harvested from MPS I mice and derived into macrophages. MPS I macrophages were incubated with rhIDU with or without immunized rabbit serum, pre-immune serum, murine Fc receptor blocking agent and mannose 6-phosphate. We evaluated two types of immunized rabbit serum: one that completely abolished rhIDU uptake into human MPS I fibroblasts (inhibiting serum) and one that only partially (30%) inhibited rhIDU uptake into human fibroblasts (partially-inhibiting serum). The uptake of rhIDU in the presence of serum containing rhIDU antibodies was enhanced into MPS I mouse macrophages when both inhibiting and partially-inhibiting serum was applied. Uptake of rhIDU per cell was higher in macrophages when immune serum than was uptake of rhIDU into human fibroblasts normalized per cell. Treatment with mannose 6-phosphate or Fc receptor block partially inhibited the uptake in macrophages. Our study demonstrated that even a low-level humoral immune response against rhIDU, which only partially inhibits rhIDU uptake into fibroblasts in vitro, nevertheless might alter its tissue distribution in vivo. At the cellular level, antibody-positive mice show reduced rhIDU distribution in hepatocytes, while distribution to tissue macrophages is maintained. In vitro, immunized serum reduced rhIDU uptake into human fibroblasts but increased uptake into murine macrophages. Our findings imply that the altered tissue distribution of rhIDU caused by anti-rhIDU antibodies is partly due to reduced uptake into fibroblasts and partly due to enhanced uptake into tissue macrophages. These results imply that functional immune assays of rhIDU uptake in vitro into fibroblasts may not completely predict the impact of the humoral immune response against enzyme replacement therapy.