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Multiple combinations of CeO2-Dolomite as fillers and Intumescent Flame Retardant (IFR) ingredients were used to optimize the intumescent coatings designed for I-beam steel substrates. The influenced of fillers and various combinations of flame-retardants on the fire protective performance of the coatings were evaluated using vertical Bunsen burner fire test and various characterization techniques. Formula C and Formula F having 1:1 and 2:2 CeO2-Dolomite ratio, obtained the lowest substrate temperature around 150oC and 150.4oC, respectively after 90 minutes fire exposure. Also, the morphological structures of intumescent char observed by SEM-EDX, demonstrated that Formula C and Formula F stimulated the formation of homogeneous and more compacted surface structure. X-ray photoelectron spectroscopy (XPS) provide the binding energies of C and O constituents, it was observed that [-(C2H4)n-] was the most important free radical as it could promote the formation of aromatic carbon chain in the char surface. Finally, the findings of this study revealed that the selection of appropriate fillers and combinations of flame-retardant ingredients significantly influenced the morphological structure of the char layer, of which, Formula C and Formula F produced a char with higher thermal stability, resulting to a more fire resistive IFR coating during fire exposure.

Male life history strategies are regulated by the neuroendocrine system. Testosterone (T) and cortisol regulate male behaviors including parenting and facilitate managing tradeoffs at key transitions in development such as first-time fatherhood. Both hormones demonstrate marked fluctuations in the postnatal period, and this presents an opportunity to investigate the role of T and cortisol in postpartum depressive symptoms-comparably less studied in fathers than in mothers in the evolutionary literature. Prior work on depressive symptoms has yet to integrate insights from the "dual hormone hypothesis (DHH)" which has focused on how T and cortisol interact to jointly regulate traits associated with dominance and status-seeking (i.e., mating effort) but has yet to be included in models of parenting effort. In this research, we use secondary data to investigate the relationship between DHH and traits ostensibly opposed to status seeking (i.e., depressive symptoms). First-time fathers ( n  = 193) provided morning saliva samples 10 months following parturition and reported on the presence of depressive symptoms (BDI-II). Responses were decomposed into three factors: cognitive, affective, and somatic. Using hybrid latent variable structural equation modeling, we did not find evidence that T predicted variability in cognitive, affective, or somatic depressive symptom factors. We found a null effect for cortisol as well. Finally, we could not find evidence that the DHH variable (T × cortisol interaction) predicted any variability in cognitive, affective, or somatic depressive symptoms. While we did not find evidence to support our hypotheses using a secondary data set, this study contributes to research on the neuroendocrinology of depression in fathers. Discussion focuses on the limitations of sample demographics, timing of saliva and self-report collection, and the lack of extant theory specific to paternal postpartum depression., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Kotov and Corpuz.)

Across the lifespan, males negotiate the tradeoff between current and future reproduction. From a life history theory (LHT) perspective, resources invested into earlier reproduction pose a cost to later reproduction. The age of sexual debut is a commonplace measure of sexual maturation. However, in males, thorarche (age of first ejaculation) and years from thorarche to age of first reproduction both represent milestones related to reproductive timing. A fundamental prediction from LHT is that earlier sexual maturation-a "quantity" strategy-predicts decreased levels of care per offspring. In the current study, we test this straightforward relationship looking specifically at a father's investment of time. In a sample of first-time fathers, we measured the amount of time spent with their 9-to-12-month infants longitudinally using an experience sampling method (ESM)-an ecologically valid method of collecting self-report data on fathers' use of time Fathers contributed data on their time allocation across a 12-week period. They reported on ages of sexual debut, thorarche, and the years between thorarche and first reproduction (i.e., current age) was calculated. Only age of sexual debut had a relationship with time allocated toward infants. Importantly however, this effect was in a direction opposite of our LHT derived hypothesis. Males with earlier sexual debut spent more time with their infants. Discussion focuses on the potential contributions to this finding and limitations related to small effect size, methods and measurement, and sample demographics., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Corpuz, Kotov and Donovan.)

Parkinson's disease (PD), a neurodegenerative disease affecting dopaminergic (DA) neurons, is characterized by decline of motor function and cognition. Dopaminergic cell loss is associated with accumulation of toxic alpha synuclein aggregates. As DA neuron death occurs late in the disease, therapeutics that block the spread of alpha synuclein may offer functional benefit and delay disease progression. To test this hypothesis, we generated antibodies to the C terminal region of synuclein with high nanomolar affinity and characterized them in in vitro and in vivo models of spread. Interestingly, we found that only antibodies with high affinity to the distal most portion of the C-terminus robustly reduced uptake of alpha synuclein preformed fibrils (PFF) and accumulation of phospho (S129) alpha synuclein in cell culture. Additionally, the antibody treatment blocked the spread of phospho (S129) alpha synuclein associated-pathology in a mouse model of synucleinopathy. Blockade of neuronal PFF uptake by different antibodies was more predictive of in vivo activity than their binding potency to monomeric or oligomeric forms of alpha synuclein. These data demonstrate that antibodies directed to the C-terminus of the alpha synuclein have differential effects on target engagement and efficacy. Furthermore, our data provides additional support for the development of alpha synuclein antibodies as a therapeutic strategy for PD patients., (Copyright © 2022. Published by Elsevier Inc.)

Background: Over the past decade, human Interleukin 33 (hIL-33) has emerged as a key contributor to the pathogenesis of numerous inflammatory diseases. Despite the existence of several commercial hIL-33 assays spanning multiple platform technologies, their ability to provide accurate hIL-33 concentration measurements and to differentiate between active (reduced) and inactive (oxidized) hIL-33 in various matrices remains uncertain. This is especially true for lower sample volumes, matrices with low hIL-33 concentrations, and matrices with elevated levels of soluble Interleukin 1 Receptor-Like 1 (sST2), an inactive form of ST2 that competes with membrane bound ST2 for hIL-33 binding., Results: We tested the performance of several commercially available hIL-33 detection assays in various human matrices and found that most of these assays lacked the sensitivity to accurately detect reduced hIL-33 at biologically relevant levels (sub-to-low pg/mL), especially in the presence of human sST2 (hsST2), and/or lacked sufficient target specificity. To address this, we developed and validated a sensitive and specific enzyme-linked immunosorbent assay (ELISA) capable of detecting reduced and total hIL-33 levels even in the presence of high concentrations of sST2. By incorporating the immuno-polymerase chain reaction (iPCR) platform, we further increased the sensitivity of this assay for the reduced form of hIL-33 by ~ 52-fold. Using this hIL-33 iPCR assay, we detected hIL-33 in postmortem human vitreous humor (VH) samples from donors with age-related macular degeneration (AMD) and found significantly increased hIL-33 levels when compared to control individuals. No statistically significant difference was observed in aqueous humor (AH) from AMD donors nor in plasma and nasosorption fluid (NF) from asthma patients compared to control individuals., Conclusions: Unlike existing commercial hIL-33 assays, our hIL-33 bioassays are highly sensitive and specific and can accurately quantify hIL-33 in various human clinical matrices, including those with high levels of hsST2. Our results provide a proof of concept of the utility of these assays in clinical trials targeting the hIL-33/hST2 pathway., (© 2021. The Author(s).)

Transforming growth factor-β (TGFβ) is a key driver of fibrogenesis. Three TGFβ isoforms (TGFβ1, TGFβ2, and TGFβ3) in mammals have distinct functions in embryonic development; however, the postnatal pathological roles and activation mechanisms of TGFβ2 and TGFβ3 have not been well characterized. Here, we show that the latent forms of TGFβ2 and TGFβ3 can be activated by integrin-independent mechanisms and have lower activation thresholds compared to TGFβ1. Unlike TGFB1 , TGFB2 and TGFB3 expression is increased in human lung and liver fibrotic tissues compared to healthy control tissues. Thus, TGFβ2 and TGFβ3 may play a pathological role in fibrosis. Inducible conditional knockout mice and anti-TGFβ isoform-selective antibodies demonstrated that TGFβ2 and TGFβ3 are independently involved in mouse fibrosis models in vivo, and selective TGFβ2 and TGFβ3 inhibition does not lead to the increased inflammation observed with pan-TGFβ isoform inhibition. A cocrystal structure of a TGFβ2-anti-TGFβ2/3 antibody complex reveals an allosteric isoform-selective inhibitory mechanism. Therefore, inhibiting TGFβ2 and/or TGFβ3 while sparing TGFβ1 may alleviate fibrosis without toxicity concerns associated with pan-TGFβ blockade., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Myeloid cells encounter stromal cells and their matrix determinants on a continual basis during their residence in any given organ. Here, we examined the impact of the collagen receptor LAIR1 on myeloid cell homeostasis and function. LAIR1 was highly expressed in the myeloid lineage and enriched in non-classical monocytes. Proteomic definition of the LAIR1 interactome identified stromal factor Colec12 as a high-affinity LAIR1 ligand. Proteomic profiling of LAIR1 signaling triggered by Collagen1 and Colec12 highlighted pathways associated with survival, proliferation, and differentiation. Lair1 -/- mice had reduced frequencies of Ly6C - monocytes, which were associated with altered proliferation and apoptosis of non-classical monocytes from bone marrow and altered heterogeneity of interstitial macrophages in lung. Myeloid-specific LAIR1 deficiency promoted metastatic growth in a melanoma model and LAIR1 expression associated with improved clinical outcomes in human metastatic melanoma. Thus, monocytes and macrophages rely on LAIR1 sensing of stromal determinants for fitness and function, with relevance in homeostasis and disease., Competing Interests: Declarations of interests All authors are stockholders of Genentech/Roche except B.H., A.W., E.V., and S.C., (Copyright © 2021 Elsevier Inc. All rights reserved.)

In human males, testosterone (T) decreases in the period following the birth of offspring. This decline has been widely interpreted as a facultative neuroendocrine response that facilitates parenting effort. Conversely, research on if (or when) this decline in T would be followed by an eventual recovery and subsequent shift away from parenting effort is lacking. In a U.S. community sample of 225 males transitioning to first-time fatherhood, we measured T at three occasions: third trimester, infant 3 months postnatal, and infant 9-10 months postnatal. Using a piecewise latent growth curve model (GCM), we detected a T rebound from when infants were 3 months old to when infants were 9-10 months old. The slope of this rebound was able to predict paternal care using two distinct measures: (a) an experience sampling method (ESM) that gathered data on paternal time allocation over the course of the study period and (b) independent coders rating fathers for the quality of paternal care during a structured task designed to elicit an infant fear response. As predicted, the more accelerated one's T rebound (slope), the less time fathers invested in their infants across the study period. However, we found a positive relationship between T rebound and quality of paternal care during a challenging activity. Discussion will focus on nuanced reasons that contribute to these findings as well as speculate on the ultimate function of a human paternal T rebound., (© 2020 Wiley Periodicals LLC.)

The postnatal period is a time of increased brain development and plasticity which have enduring influences on brain and behavior. Infants demonstrate a transient surge in testosterone (T) during development referred to as "mini puberty". The utility of studying mini puberty in psychobiology has only recently emerged. Life-history theory postulates that infants "use" the maternal environment-pre and postnatally-to calibrate growth and timing of sexual maturity. As such, variability in infant T levels is not arbitrary and can be predicted by theory. We examine the role of maternal pre- and postnatal cortisol. Using saliva samples (n = 193 dyads), we predicted that higher levels of maternal cortisol are associated with higher levels of infant T. We found only maternal postnatal cortisol had a relationship with infants' mini puberty. This relationship was in the predicted direction and remained after controlling for numerous variables. Discussion will include the potential role of mini puberty as an inflection point where systems related to growth, sexual maturation, and psychosexual behavior can be calibrated and coordinated., (© 2020 Wiley Periodicals LLC.)

Non-alcoholic Fatty Liver Disease (NAFLD) is the most common form of liver disease and is associated with metabolic dysregulation. Although G protein-coupled receptor 84 (GPR84) has been associated with inflammation, its role in metabolic regulation remains elusive. The aim of our study was to evaluate the potential of PBI-4547 for the treatment of NAFLD and to validate the role of its main target receptor, GPR84. We report that PBI-4547 is a fatty acid mimetic, acting concomitantly as a GPR84 antagonist and GPR40/GPR120 agonist. In a mouse model of diet-induced obesity, PBI-4547 treatment improved metabolic dysregulation, reduced hepatic steatosis, ballooning and NAFLD score. PBI-4547 stimulated fatty acid oxidation and induced gene expression of mitochondrial uncoupling proteins in the liver. Liver metabolomics revealed that PBI-4547 improved metabolic dysregulation induced by a high-fat diet regimen. In Gpr84 -/- mice, PBI-4547 treatment failed to improve various key NAFLD-associated parameters, as was observed in wildtype littermates. Taken together, these results highlight a detrimental role for the GPR84 receptor in the context of meta-inflammation and suggest that GPR84 antagonism via PBI-4547 may reflect a novel treatment approach for NAFLD and its related complications.

The ongoing Coronavirus disease (COVID-19) pandemic has had a devastating impact worldwide. It is unclear as to what one expects during the "post-peak" and "post-pandemic" periods in terms of: (1) continued adherence to precautionary measures (e.g., wearing a mask) and (2) behaviors during these periods pertaining to widespread (anticipated) medical solutions that can buffer subsequent waves (e.g., vaccination and donating plasma). In this study, we examine predictors of individual differences in attitudes and behaviors with regard to the COVID-19 pandemic and the months moving forward. Of the factors that contribute to how one might navigate the pandemic - a source of elevated environmental threat - life history orientation may play a crucial role. In this study, participants ( n = 209) indicated their agreement with items on attitudes toward COVID-19 precautions and medical solutions that can buffer subsequent waves. In all models, we found significant positive relationships between one's slow life history orientation and their self-reported adherence to precautions and endorsement of medical solutions. This effect was detectable even after controlling for factors related to political conservatism and personal experience with deleterious events as a result of the pandemic. Discussion includes reflection on the main finding, demographic variables, as well as the relationships uncovered among the modeled covariates (e.g., social conservatism, political conservatism)., (Copyright © 2020 Corpuz, D’Alessandro, Adeyemo, Jankowski and Kandalaft.)

Male testosterone (T) decreases in response to childbirth. Longitudinal support for this has come from samples across cultures. In this study, we look at individual differences in this phenomenon. Utilizing a sample of U.S. fathers, we employ life history theory to investigate the influence of a father's early experience on his neuroendocrine response to fatherhood. We conducted three home visits (n = 226 fathers) from the third trimester of pregnancy to when infants were 10 months old. In this sample, T declined from the third trimester of (a partner's) pregnancy to the early months of the postnatal period. T recovered to pre-birth levels by the time infants reached 10 months old. We did not find any evidence that one's subjective experience of their early environment could account for any meaningful variability in T calibration. Objective, "event" measures of early harshness (i.e., death of a sibling/friend) and unpredictability (i.e., parent upheaval) each uniquely predicted a younger age of sexual debut. Neither harshness nor unpredictability had any (direct or indirect) effects on T calibration. Age of sexual debut did predict the rate of T recovery from 3 to 10 months postnatal. The younger one's sexual debut, the more accelerated their T ascent during this period. We discuss the potential reasons for, and implications of our mixed results., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Severe asthma patients with low type 2 inflammation derive less clinical benefit from therapies targeting type 2 cytokines and represent an unmet need. We show that mast cell tryptase is elevated in severe asthma patients independent of type 2 biomarker status. Active β-tryptase allele count correlates with blood tryptase levels, and asthma patients carrying more active alleles benefit less from anti-IgE treatment. We generated a noncompetitive inhibitory antibody against human β-tryptase, which dissociates active tetramers into inactive monomers. A 2.15 Å crystal structure of a β-tryptase/antibody complex coupled with biochemical studies reveal the molecular basis for allosteric destabilization of small and large interfaces required for tetramerization. This anti-tryptase antibody potently blocks tryptase enzymatic activity in a humanized mouse model, reducing IgE-mediated systemic anaphylaxis, and inhibits airway tryptase in Ascaris-sensitized cynomolgus monkeys with favorable pharmacokinetics. These data provide a foundation for developing anti-tryptase as a clinical therapy for severe asthma., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Traditional hybridoma and B cell cloning antibody discovery platforms have inherent limits in immune repertoire sampling depth. One consequence is that monoclonal antibody (mAb) leads often lack the necessary affinity for therapeutic applications, thus requiring labor-intensive and time-consuming affinity in vitro engineering optimization steps. Here, we show that high-affinity variants of mouse-derived mAbs can be rapidly obtained by testing of somatic sequence variants obtained by deep sequencing of antibody variable regions in immune repertories from immunized mice, even with a relatively sparse sampling of sequence variants from large sequence datasets. Affinity improvements can be achieved for mAbs with a wide range of affinities. The optimized antibody variants derived from immune repertoire mining have no detectable in vitro off-target binding and have in vivo clearance comparable to the parental mAbs, essential properties in therapeutic antibody leads. As generation of antibody variants in vitro is replaced by mining of variants generated in vivo, the procedure can be applied to rapidly identify affinity-optimized mAb variants.

FGF21 is a stress-induced hormone with potent anti-obesity, insulin-sensitizing, and hepatoprotective properties. Although proteolytic cleavage of recombinant human FGF21 in preclinical species has been observed previously, the regulation of endogenously produced FGF21 is not well understood. Here we identify fibroblast activation protein (FAP) as the enzyme that cleaves and inactivates human FGF21. A selective chemical inhibitor, immunodepletion, or genetic deletion of Fap stabilized recombinant human FGF21 in serum. In addition, administration of a selective FAP inhibitor acutely increased circulating intact FGF21 levels in cynomolgus monkeys. On the basis of our findings, we propose selective FAP inhibition as a potential therapeutic approach to increase endogenous FGF21 activity for the treatment of obesity, type 2 diabetes, non-alcoholic steatohepatitis, and related metabolic disorders., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)

Interferons-alpha (IFN-α) are the expressed gene products comprising thirteen type I interferons with protein pairwise sequence similarities in the 77-96% range. Three other widely expressed human type I interferons, IFN-β, IFN-κ and IFN-ω have sequences 29-33%, 29-32% and 56-60% similar to the IFN-αs, respectively. Type I interferons act on immune cells by producing subtly different immune-modulatory effects upon binding to the extracellular domains of a heterodimeric cell-surface receptor composed of IFNAR1 and IFNAR2, most notably anti-viral effects. IFN-α has been used to treat infection by hepatitis-virus type C (HCV) and a correlation between hyperactivity of IFN-α-induced signaling and systemic lupus erythematosis (SLE), or lupus, has been noted. Anti-IFN-α antibodies including rontalizumab have been under clinical study for the treatment of lupus. To better understand the rontalizumab mechanism of action and specificity, we determined the X-ray crystal structure of the Fab fragment of rontalizumab bound to human IFN-α2 at 3Å resolution and find substantial overlap of the antibody and IFNA2 epitopes on IFN-α2., (© 2015 The Protein Society.)

This study assesses the combined effects of children's early medical risk (e.g., preterm status) and parental investment levels (time spent in provision of care to target children as opposed to other family members) on children's response to novel, potentially distressing stimuli. While engaged in play activities, children were exposed to stimuli that were either neutral (a speaker on television with a calm voice) or threatening (a speaker with an angry voice). A significant interaction between children's risk status and parental investment was found only for threatening stimuli. High-risk children with high-investing parents showed high visual engagement with potentially threatening responses, whereas high-risk children with low-investing parents were more likely to show visual avoidance. No comparable effects were found for low-risk children. Findings were interpreted as showing that high-risk children with a history of high parental investment are more likely to attend to potentially threatening events, an adaptive response in the presence of reliable support., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Purpose: Our goal was to develop a potent humanized antibody against mouse/human CXCL12. This report summarized its in vitro and in vivo activities., Experimental Design: Cell surface binding and cell migration assays were used to select neutralizing hamster antibodies, followed by testing in several animal models. Monoclonal antibody (mAb) 30D8 was selected for humanization based on its in vitro and in vivo activities., Results: 30D8, a hamster antibody against mouse and human CXCL12α, CXCL12β, and CXCL12γ, was shown to dose-dependently block CXCL12α binding to CXCR4 and CXCR7, and CXCL12α-induced Jurkat cell migration in vitro. Inhibition of primary tumor growth and/or metastasis was observed in several models. 30D8 alone significantly ameliorated arthritis in a mouse collagen-induced arthritis model (CIA). Combination with a TNF-α antagonist was additive. In addition, 30D8 inhibited 50% of laser-induced choroidal neovascularization (CNV) in mice. Humanized 30D8 (hu30D8) showed similar in vitro and in vivo activities as the parental hamster antibody. A crystal structure of the hu30D8 Fab/CXCL12α complex in combination with mutational analysis revealed a "hot spot" around residues Asn(44)/Asn(45) of CXCL12α and part of the RFFESH region required for CXCL12α binding to CXCR4 and CXCR7. Finally, hu30D8 exhibited fast clearance in cynomolgus monkeys but not in rats., Conclusion: CXCL12 is an attractive target for treatment of cancer and inflammation-related diseases; hu30D8 is suitable for testing this hypothesis in humans., (©2013 AACR.)

Multiple Sclerosis (MS) is a neurodegenerative autoimmune disorder caused by chronic inflammation and demyelination within the central nervous system (CNS). Clinical studies in MS patients have demonstrated efficacy with B cell targeted therapies such as anti-CD20. However, the exact role that B cells play in the disease process is unclear. Activation Induced cytidine deaminase (AID) is an essential enzyme for the processes of antibody affinity maturation and isotype switching. To evaluate the impact of affinity maturation and isotype switching, we have interrogated the effect of AID-deficiency in an animal model of MS. Here, we show that the severity of experimental autoimmune encephalomyelitis (EAE) induced by the extracellular domain of human myelin oligodendrocyte glycoprotein (MOG1-125) is significantly reduced in Aicda deficient mice, which, unlike wild-type mice, lack serum IgG to myelin associated antigens. MOG specific T cell responses are comparable between wild-type and Aicda knockout mice suggesting an active role for antigen experienced B cells. Thus affinity maturation and/or class switching are critical processes in the pathogenesis of EAE.

Mothers of medically at-risk infants were randomly assigned to a Healthy Start intervention (HV) or a cognitive reframing intervention (HV+). Outcome measures were taken at the conclusion of the intervention (1 year) and at the 3-year follow-up visit. At age 3, children in the HV+ condition (in comparison with those in the HV condition) showed fewer aggression problems (as measured by the Child Behavior Checklist). Maternal emotional unavailability (as measured by combined scores on the Beck Depression Inventory and avoidance items on the Conflict Tactics Scale) at the 1-year visit mediated the effects of the intervention on children's aggression at age 3. Findings suggest that an early, cognitively based intervention may lead to reduced child aggression as a result of increased maternal social-emotional availability within the caregiving relationship., (PsycINFO Database Record (c) 2012 APA, all rights reserved.)

This study seeks to ascertain whether a culturally tailored art calendar could improve participation in cancer screening activities. We conducted a randomized, controlled calendar mail-out in which a Native art calendar was sent by first class mail to 5,633 patients seen at an urban American Indian clinic during the prior 2 years. Using random assignment, half of the patients were mailed a "message" calendar with screening information and reminders on breast, colorectal, lung, and prostate cancer; the other half received a calendar without messages. The receipt of cancer screening services was ascertained through chart abstraction in the following 15 months. In total, 5,363 observations (health messages n = 2,695; no messages n = 2,668) were analyzed. The calendar with health messages did not result in increased receipt of any cancer-related prevention outcome compared to the calendar without health messages. We solicited clinic input to create a culturally appropriate visual intervention to increase cancer screening in a vulnerable, underserved urban population. Our results suggest that printed materials with health messages are likely too weak an intervention to produce the desired behavioral outcomes in cancer screening.

The transmembrane serine protease hepsin is one of the most highly upregulated genes in prostate cancer. Here, we investigated its tumor-promoting activity by use of a mouse orthotopic prostate cancer model. First, we compared the tumor growth of low hepsin-expressing LnCaP-17 cells with hepsin-overexpressing LnCaP-34 cells. After implantation of cells into the left anterior prostate lobe, LnCaP-34 tumors not only grew faster based on increased serum prostate-specific antigen levels but also metastasized to local lymph nodes and, most remarkably, invaded the contralateral side of the prostate at a rate of 100% compared with only 18% for LnCaP-17 tumors. The increased tumor growth was not due to nonspecific gene expression changes and was not predicted from the unaltered in vitro growth and invasion of LnCaP-34 cells. A likely explanation is that the in vivo effects of hepsin were mediated by specific hepsin substrates present in the tumor stroma. In a second study, mice bearing LnCaP-34 tumors were treated with a PEGylated form of Kunitz domain-1, a potent hepsin active site inhibitor derived from hepatocyte growth factor activator inhibitor-1 (K(i)(app) 0.30 +/- 0.02 nmol/L). Treatment of established tumors with PEGylated Kunitz domain-1 decreased contralateral prostate invasion (46% weight reduction) and lymph node metastasis (50% inhibition). Moreover, serum prostate-specific antigen level remained reduced during the entire treatment period, reaching a maximal reduction of 76% after 5 weeks of dosing. The findings show that hepsin promotes invasive prostate tumor growth and metastasis and suggest that active site-directed hepsin inhibition could be effective in prostate cancer therapy.

VEGF-A is important in tumor angiogenesis, and a humanized anti-VEGF-A monoclonal antibody (bevacizumab) has been approved by the FDA as a treatment for metastatic colorectal and nonsquamous, non-small-cell lung cancer in combination with chemotherapy. However, contributions of both tumor- and stromal-cell derived VEGF-A to vascularization of human tumors grown in immunodeficient mice hindered direct comparison between the pharmacological effects of anti-VEGF antibodies with different abilities to block host VEGF. Therefore, by gene replacement technology, we engineered mice to express a humanized form of VEGF-A (hum-X VEGF) that is recognized by many anti-VEGF antibodies and has biochemical and biological properties comparable with WT mouse and human VEGF-A. The hum-X VEGF mouse model was then used to compare the activity and safety of a panel of VEGF Mabs with different affinities for VEGF-A. Although in vitro studies clearly showed a correlation between binding affinity and potency at blocking endothelial cell proliferation stimulated by VEGF, in vivo experiments failed to document any consistent correlation between antibody affinity and the ability to inhibit tumor growth and angiogenesis in most animal models. However, higher-affinity antibodies were more likely to result in glomerulosclerosis during long-term treatment.

Chemokines play an important role in the immune system by regulating cell trafficking in homeostasis and inflammation. In this study, we report the identification and characterization of a novel cytokine-like protein, DMC (dendritic cell and monocyte chemokine-like protein), which attracts dendritic cells and monocytes. The key to the identification of this putative new chemokine was the application of threading techniques to its uncharacterized sequence. Based on our studies, DMC is predicted to have an IL-8-like chemokine fold and to be structurally and functionally related to CXCL8 and CXCL14. Consistent with our predictions, DMC induces migration of monocytes and immature dendritic cells. Expression studies show that DMC is constitutively expressed in lung, suggesting a potential role for DMC in recruiting monocytes and dendritic cells from blood into lung parenchyma.

Hormonal control of metabolic rate can be important in regulating the imbalance between energy intake and expenditure that underlies the development of obesity. In mice fed a high-fat diet, human fibroblast growth factor 19 (FGF19) increased metabolic rate [1.53 +/- 0.06 liters O(2)/h.kg(0.75) (vehicle) vs. 1.93 +/- 0.05 liters O(2)/h.kg(0.75) (FGF19); P < 0.001] and decreased respiratory quotient [0.82 +/- 0.01 (vehicle) vs. 0.80 +/- 0.01 (FGF19); P < 0.05]. In contrast to the vehicle-treated mice that gained weight (0.14 +/- 0.05 g/mouse.d), FGF19-treated mice lost weight (-0.13 +/- 0.03 g/mouse.d; P < 0.001) without a significant change in food intake. Furthermore, in addition to a reduction in weight gain, treatment with FGF19 prevented or reversed the diabetes that develops in mice made obese by genetic ablation of brown adipose tissue or genetic absence of leptin. To explore the mechanisms underlying the FGF19-mediated increase in metabolic rate, we profiled the FGF19-induced gene expression changes in the liver and brown fat. In brown adipose tissue, chronic exposure to FGF19 led to a gene expression profile that is consistent with activation of this tissue. We also found that FGF19 acutely increased liver expression of the leptin receptor (1.8-fold; P < 0.05) and decreased the expression of acetyl coenzyme A carboxylase 2 (0.6-fold; P < 0.05). The gene expression changes were consistent with the experimentally determined increase in fat oxidation and decrease in liver triglycerides. Thus, FGF19 is able to increase metabolic rate concurrently with an increase in fatty acid oxidation.

Genetic defects in the Wnt-1 signaling pathway contribute to human tumor progression and are especially prevalent in colorectal cancer. We screened mouse C57MG cells to isolate mRNAs induced by Wnt-1 and identified Stra6, an mRNA known to be up-regulated by retinoic acid. Up-regulation of Stra6 mRNA was also observed in hyperplastic mammary tissue and mammary gland tumors from transgenic mice expressing Wnt-1 and in human tumors that frequently harbor defects in Wnt-1 signaling. Stimulation of C57MG cells with retinoic acid plus Wnt-1 resulted in expression of Stra6 transcript to levels greatly exceeding that observed with either stimulus alone. This synergy could be explained in part by the up-regulation of retinoic acid receptor-gamma that was observed in response to Wnt-1 signaling. Accordingly, treatment of human colorectal cancer cell lines with retinoic acid resulted in the up-regulation of Stra6 mRNA and accumulation of Stra6 protein at the cell membrane. The data support a model in which Wnt-1 signaling synergizes with retinoids to activate retinoic acid receptor-gamma-responsive genes in human cancers.

We report identification of interleukin (IL)-17E, a novel member of the IL-17 family of cytokines. IL-17E is a ligand for the recently identified protein termed EVI27/IL-17BR, which we term IL-17 receptor homolog 1 (IL-17Rh1) in light of the multiple reported ligand-receptor relationships. Murine EVI27 was identified through its location at a common site of retroviral integration in BXH2 murine myeloid leukemias. IL-17Rh1 shows highest level expression in kidney with moderate expression in multiple other organs, whereas IL-17E mRNA was detected at very low levels in several peripheral tissues. IL-17E induces activation of NF-kappaB and stimulates production of the proinflammatory chemokine IL-8.

The stability of captopril in powder papers under three different storage conditions was determined. Captopril 12.5-mg tablets were triturated with lactose to a final concentration of 2 mg of captopril in 100 mg of powder. A total of 240 powder papers were prepared and stored in class "A" prescription vials (80 papers), 002G plastic zip-lock bags (80 papers), and Moisture Proof Barrier Bags (80 papers). Immediately after preparation and at 1, 2, 3, 4, 8, 12, and 24 weeks of storage at room temperature, powder papers under each storage condition were reweighed and the contents were assayed for captopril concentration by a stability-indicating high-performance liquid chromatographic method. More than 90% of the initial captopril concentration was retained under all storage conditions during the first 12 weeks of the study. Captopril disulfide, a degradation product, was detected in one sample stored in a plastic zip-lock bag at 24 weeks. Captopril was stable for the entire 24-week period in powder papers stored in either the class A prescription vial or the Moisture Proof Barrier Bag. Captopril in powder papers is stable for at least 12 weeks when stored at room temperature under all three storage conditions.