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Early Time Points (early + time_point)
Selected AbstractsEffectiveness of Microporous Polysaccharide Hemospheres for Achieving Hemostasis in Mohs Micrographic SurgeryDERMATOLOGIC SURGERY, Issue 6 2004FRCPC, Stephen R. Tan MD Background. Microporous polysaccharide hemospheres consist of controlled-porosity spherical particles manufactured from bioinert plant polysaccharide. Microporous polysaccharide hemospheres facilitate hemostasis by rapidly absorbing the fluid component of blood, concentrating platelets and clotting factors to accelerate blood clotting. Objective. The objective was to compare a microporous polysaccharide hemosphere bandage and electrocautery in achieving hemostasis. Methods. Twenty-four patients with a total of 48 stages of Mohs micrographic surgery were included. Patients were stratified by whether or not they were taking anticoagulant medications. Within each group, patients were randomized to receive either the microporous polysaccharide hemosphere bandage or electrocautery. Outcomes included bleeding through the dressing (early time point) and active bleeding upon dressing removal (late time point). Results. Nineteen patients not taking anticoagulants had 40 stages, of which 18 received the study bandage and 22 received electrocautery. The remaining 5 patients on anticoagulants had 8 stages, of which 4 received the study bandage and 4 received electrocautery. In both total and subgroup analysis, there was a higher incidence of bleeding through the dressing with the study bandage (p<0.05), but no increase in the incidence of active bleeding upon dressing removal (p>0.05). Conclusion. The microporous polysaccharide hemosphere study bandage had an increased incidence of bleeding through the dressing compared to electrocautery, but did not have an increased incidence of active bleeding upon dressing removal. [source] Time course analysis of gene expression during light-induced photoreceptor cell death and regeneration in albino zebrafishDEVELOPMENTAL NEUROBIOLOGY, Issue 8 2007Sean C. Kassen Abstract Constant intense light causes apoptosis of rod and cone photoreceptors in adult albino zebrafish. The photoreceptors subsequently regenerate from proliferating inner nuclear layer (INL) progenitor cells that migrate to the outer nuclear layer (ONL) and differentiate into rods and cones. To identify gene expression changes during this photoreceptor regeneration response, a microarray analysis was performed at five time points during the light treatment. The time course included an early time point during photoreceptor death (16 h), later time points during progenitor cell proliferation and migration (31, 51, and 68 h) and a 96 h time point, which likely corresponds to the initial photoreceptor differentiation. Mean expression values for each gene were calculated at each time point relative to the control (0 h light exposure) and statistical analysis by one-way ANOVA identified 4567 genes exhibiting significant changes in gene expression along the time course. The genes within this data set were clustered based on their temporal expression patterns and proposed functions. Quantitative real-time PCR validated the microarray expression profiles for selected genes, including stat3 whose expression increased markedly during the light exposure. Based on immunoblots, both total and activated Stat3 protein expression also increased during the light treatment. Immunolocalization of Stat3 on retinal tissue sections demonstrated increased expression in photoreceptors and Müller glia by 16 h of light exposure. Some of the Stat3-positive Müller cells expressed PCNA at 31 h, suggesting that Stat3 may play a role in signaling a subset of Müller cells to proliferate during the regeneration response. © 2007 Wiley Periodicals, Inc. Develop Neurobiol, 2007. [source] Microcell-mediated transfer of chromosome 4 into HeLa cells suppresses telomerase activityGENES, CHROMOSOMES AND CANCER, Issue 2 2001Claudia Backsch Telomerase activity can be detected in most human cancers and immortal cell lines. In contrast, the lack of telomerase activity in normal diploid fibroblasts has been correlated with progressive reduction of telomere lengths to critically short sizes followed by the cessation of cell division and the onset of senescence. Several investigators have provided evidence for the localization of a telomerase suppressor gene on chromosome 3. The aim of our study was to determine whether other chromosomes are involved in telomerase repression. Beside human chromosome 3 (serving as positive control), chromosomes 4, 6, and 11 were introduced into HeLa cells via microcell-mediated chromosome transfer. Telomerase activity from different hybrid cell lysates was determined at an early time point after fusion using a Telomerase ELISA kit. Strong repression of telomerase activity was only found in a subset of HeLa hybrids in which chromosome 3 or chromosome 4 had been introduced. Telomerase suppression induced by chromosome 3 or 4 transfer was paralleled by a high frequency (30% or 43%, respectively) of a senescent-like phenotype. Chromosomes 6 and 11, the functional loss of which is also implicated in cervical cancer, had no effect. These results indicate that normal human chromosomes 3 and 4 carry a gene or genes that suppress telomerase activity and induce cellular senescence in HeLa cells.©2001 Wiley-Liss, Inc. [source] In Vivo and In Vitro Evidence of the Involvement of CXCL1, a Keratinocyte-Derived Chemokine, in Equine LaminitisJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 5 2009R.R. Faleiros Background: C-X-C motif ligand 1 (CXCL1) is an important chemokine of epithelial origin in rodents and humans. Objectives: To assess in vivo and in vitro the regulation of CXCL1 in equine laminitis. Animals: Twenty adult horses. Methods: Real-time quantitative polymerase chain reaction (PCR) was used to assess expression of CXCL1 in samples of laminae, liver, skin, and lung from the black walnut extract (BWE) model of laminitis, and in cultured equine epithelial cells (EpCs). Tissue was obtained from control animals (CON, n = 5), and at 1.5 hours (early time point [ETP] group, n = 5), at the onset of leukopenia (developmental time point [DTP] group, n = 5), and at the onset of lameness (LAM group, n = 5) after BWE administration. EpCs were exposed to Toll-like/Nod receptor ligands, oxidative stress agents, and reduced atmospheric oxygen (3%). In situ PCR was used to localize the laminar cell types undergoing CXCL1 mRNA expression. Results: Increases in laminar CXCL1 mRNA concentrations occurred in the ETP (163-fold [P= .0001]) and DTP groups (21-fold [P= .005]). Smaller increases in CXCL1 expression occurred in other tissues and organs. In cultured EpCs, increases (P < .05) in CXCL1 mRNA concentration occurred after exposure to lipopolysaccharide (LPS [28-fold]), xanthine/xanthine oxidase (3.5-fold), and H2O2 (2-fold). Hypoxia enhanced the LPS-induced increase in CXCL1 mRNA (P= .007). CXCL1 gene expression was localized to laminar EpCs, endothelial cells, and emigrating leukocytes. Conclusion and Clinical Importance: These findings indicate that CXCL1 plays an early and possibly initiating role in neutrophil accumulation in the BWE laminitis model, and that laminar keratinocytes are an important source of this chemokine. New therapies using chemokine receptor antagonists may be indicated. [source] Proteomic and transcriptomic study on the action of a cytotoxic saponin (Polyphyllin D): Induction of endoplasmic reticulum stress and mitochondria-mediated apoptotic pathwaysPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 15 2008Fung-Ming Siu Abstract Polyphyllin D (PD) is a potent cytotoxic saponin found in Paris polyphylla. In the present study, bioinformatic, proteomic and transcriptomic analyses were performed to study the mechanisms of action of PD on human nonsmall cell lung cancer (NSCLC) cell line (NCI-H460). Using a gene expression-based bioinformatic tool (connectivity map), PD was identified as a potential ER stress inducer. Our proteomic and transcriptomic analyses revealed that PD treatment led to upregulation of typical ER stress-related proteins/genes including glucose-regulated protein 78 (BiP/GRP78) and protein disulfide isomerase (PDI). In particular, elevated expression of C/EBP homologous transcription factor (chop) and activation of caspase-4 occurred at early time point (8,h) of PD treatment, signifying an initial ER stress-mediated apoptosis. Induction of tumor suppressor p53, disruption of mitochondrial membrane, activation of caspase-9 and caspase-3 were detected upon prolonged PD treatment. Collectively, these data revealed that PD induced the cytotoxic effect through a mechanism initiated by ER stress followed by mitochondrial apoptotic pathway. The ability of activating two major pathways of apoptosis makes PD an attractive drug lead for anticancer therapeutics. [source] Influence of DNA repair gene polymorphisms on the initial repair of MMS-induced DNA damage in human lymphocytes as measured by the alkaline comet assayENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 9 2008Charlotta Ryk Abstract We have applied the alkaline comet assay to study the functional impact of gene polymorphisms in base excision repair (APEX1 Asp148Glu, XRCC1 Arg194Trp, XRCC1 Arg399Gln) and homologous recombination repair (XRCC3 Thr241Met, NBS1 Glu185Gln), two pathways that play crucial roles in the repair of DNA damage induced by methylmethane sulphonate (MMS). We also examined the effect of polymorphisms in mismatch repair (MLH1 ,93 A/G) and nucleotide excision repair (XPD Lys751Gln) as putative negative controls based on the limited roles of these pathways in MMS-induced repair. Phytohemagglutinin-stimulated peripheral lymphocytes from 52 healthy individuals were treated with MMS and allowed to repair for 0, 15, 40, or 120 min after a 6-min washing step. DNA damage was measured as a pseudo-percentage score (comparable to % tail DNA) converted from a total visual score calculated from the distribution of cells with different degrees of damage (normal, mild, moderate and severe). The repair was faster at the beginning of the observation period than towards the end, and was not complete after 2 hr. Presence of the APEX1 148Asp, XRCC3 241Met or NBS1 185Gln alleles were significantly associated with a high pseudo-percentage score (above median) at early time points, with the APEX1 effect being most prolonged (up to 40 min after washing, odds ratio 5.6, 95% confidence interval 2.0,15.5). No significant effects were seen with the XRCC1 Arg194Trp, XRCC1 Arg399Gln, MLH1 ,93A/G and XPD Lys751Gln polymorphisms. Our results provide evidence for the functional nature of the variant alleles studied in the APEX1, XRCC3, and NBS1 genes. Environ. Mol. Mutagen., 2008. © 2008 Wiley-Liss, Inc. [source] sgk1, a member of an RNA cluster associated with cell death in a model of Parkinson's diseaseEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 2 2005Christine C. Stichel Abstract In an effort to gain deeper insight into the molecular processes underlying neurodegeneration in Parkinson's disease, we performed gene expression profiling at several early time points after MPTP-injection into old (1-year) mice. We used a PCR-based gene expression profiling method, digital expression pattern display (DEPD), a method of very high sensitivity and reproducibility, which displays almost all transcripts of a tissue. To identify cell death-associated genes, we defined clusters of differentially expressed transcripts with expression behaviour that correlated with the temporal profile of cell death progression and characterized one of these cell death clusters further. We selected one of the strongest regulated genes, the serum and glucocorticoid-regulated kinase 1 (sgk1), and validated its differential expression by Northern blot analysis, semiquantitative PCR and in situ hybridization. Up-regulation of sgk1 (i) coincides with the onset of dopaminergic cell death in both the 8-week acute and 1-year subacute MPTP models, (ii) spans the entire brain, (iii) is attenuated by the l -deprenyl-mediated inhibition of the MPTP conversion to its active metabolite MPP+ and (iv) is not induced by dehydration. This study demonstrated that the combination of the DEPD technology, clustering analysis and a detailed histopathology is a useful tool for elucidating molecular pathways in neurodegenerative diseases. [source] Surface Physiochemistry Affects Protein Adsorption to Stoichiometric and Silicate-Substituted Microporous Hydroxyapatites,ADVANCED ENGINEERING MATERIALS, Issue 4 2010Katharina Guth An important factor in the bioactivity and success of a bone-graft substitute is the nature of the adsorbed protein layer, which plays a vital role in orchestrating cell attachment and development through the presence of adhesion proteins such as fibronectin (Fn) and vitronectin (Vn). In this study, microporous hydroxyapatite (HA) and silicate-substituted hydroxyapatite (SA) discs with matched porosity and surface morphology are developed to mimic the topography found in commercial bone-graft substitutes in order to identify whether the introduction of microporosity and associated surface roughness eliminates the beneficial effect that silicate substitution has on protein adsorption. The introduction of microporosity does not abolish the relative enrichment of the protein layer that is adsorbed to the microporous SA discs, as opposed to HA, but appears to accelerate it. Fibronectin and Vn adsorption in a range of competitive environments at physiological temperatures confirm that the microporous SA discs have a greater affinity for Fn and Vn compared with HA, suggesting differences in the mechanisms behind the surface affinity to SA. Thus, development of a surface protein layer on SA and HA is likely to be dependent on the nature of the local protein environment and a combination of factors that are associated with the addition of silicate: the surface charge, the nature of the ionic species at the interface and the resultant hydrophilicity of the surface. Total protein adsorption is not found to be a good indicator of potential implant performance, particularly at early time points. [source] Analysis of chimerism during the early period after allogeneic peripheral stem cell transplantationINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 6 2001B. Gleissner As there are few reports on early evaluation of chimerism, we assessed fluorescence short tandem repeats (STR) by polymerase chain reaction (PCR) assays to analyse donor and recipient characteristics at early time points after peripheral stem cell transplantation (PBSCT). Peripheral blood of 13 patients was analysed in 1- to 2-day intervals starting from the day of PBSCT. Donor and recipient allelic patterns were determined by a commercially available multiplex STR assay that simultaneously evaluates four or five gene loci. Mixed chimerism appeared in all patients during days 1,9 after transplantation and preceded haematologic engraftment for 3,12 days. Even patients without myeloablative conditioning therapy (n=4) revealed donor allelic patterns within 1,5 days. Nine patients changed during the following days to a complete donor allelic pattern and had an uncomplicated post-transplant disease course. Four patients did not consistently retain complete donor chimerism; two of them relapsed within the next 3 months, one died from septicemia within 7 days, and the fourth, transplanted for aplastic anaemia, is still in complete remission. Overall, STR analysis using a simple and comparatively cheap multiplex system permits the detection of chimerism very early after transplantation and may provide relevant information that correlates with the clinical follow-up. [source] Community pharmacy provision of allergic rhinitis treatments: a longitudinal study of patient reported outcomeINTERNATIONAL JOURNAL OF PHARMACY PRACTICE, Issue 4 2005Dr. Hazel Sinclair Phd research fellow Objective To monitor and compare the symptoms, and reported quality of life, of two groups of people who obtained treatment for allergic rhinitis from community pharmacies (prescribed or purchased). Method Subjects were recruited by 64 community pharmacies in 2001 and followed up by postal questionnaire at four time points: five days, four weeks, eight weeks and 26 weeks. Setting Primary care: community pharmacies in Grampian, Scotland. Results Response rates: five days , 84%; four weeks , 63%; eight weeks , 59%; 26 weeks , 56%. Three hundred and twenty-four subjects completed the five-day questionnaire (138 prescribed, 186 purchased). There were no important differences between groups in socio-economic variables monitored. The commonest treatments provided were antihistamines (non-sedating: 63% prescribed, 59% purchased; sedating: 3% prescribed, 16% purchased). Despite treatment, symptoms and quality-of-life impairments remained high; the prescribed group reported higher levels of many symptoms (including asthma), and lower quality of life at early time points. Most were satisfied with their treatment and few reported unmet need for pharmacy advice (11% prescribed, 3% purchased group). Conclusion Despite high levels of patient satisfaction with allergic rhinitis treatment, symptoms and quality-of-life impairments remained high in both groups. Widespread implementation of ,allergic rhinitis and its impact on asthma' (ARIA) guidelines for physicians and for pharmacists might improve management of symptoms and quality of life of patients. [source] Increases in tumor necrosis factor-, following transient global cerebral ischemia do not contribute to neuron death in mouse hippocampusJOURNAL OF NEUROCHEMISTRY, Issue 6 2005Yuki Murakami Abstract The actions of tumor necrosis factor-, (TNF-,) produced by resident brain cells and bone marrow-derived cells in brain following a transient global ischemia were evaluated. In wild-type mice (C57Bl/6J) following 20 min ischemia with bilateral common carotid artery occlusion (BCCAo), TNF-, mRNA expression levels in the hippocampus were significantly increased at 3 h and 36 h and exhibited a biphasic expression pattern. There were no hippocampal TNF-, mRNA expression levels at early time points in either wild-type mice bone marrow transplanted (BMT)-chimeric-TNF-, gene-deficient (T/W) or TNF-, gene-deficient mice BMT-TNF-, gene-deficient mice (T/T), although TNF-, mRNA levels were detectable in T/W BMT mice at 36 h. Histopathological findings showed no intergroup differences between wild-type and TNF-, gene-deficient mice at 4 and 7 days after transient ischemia. In addition, nuclear factor-,B (NF-,B) was activated within 12 h after global cerebral ischemia, but electrophoretic mobility shift assays (EMSA) showed no intergroup differences between wild type and TNF-, gene-deficient mice. In summary, early hippocampal TNF-, mRNA expression may not be related to bone marrow-derived cells, and secondary TNF-, expression as early as 36 h after ischemia probably resulted mainly from endogenous brain cells and possibly a few bone marrow-derived cells. Although we cannot exclude the possibility of the TNF-, contribution to the physiologic changes of hippocampus after transient global ischemia, these results indicate that TNF-, does not influence the morphological changes of the hippocampal neurons under our study condition. [source] The Genetics of Acute Functional Tolerance and Initial Sensitivity to Ethanol for an Ataxia Test in the LSxSS RI StrainsALCOHOLISM, Issue 5 2000Vaughn M. Gehle Background: It has been proposed that development of tolerance to the behavioral effects of ethanol depends on the degree of impairment produced by the drug; that is, more sensitive individuals should develop greater tolerance. Tests of this hypothesis with respect to acute functional tolerance have produced contradictory results. We tested the hypothesis by examining the genetic relationship between initial sensitivity and acute functional tolerance in the LSXSS recombinant inbred mice. Methods: We tested mice for initial sensitivity to the ataxic effects of 1.75 g/kg of ethanol in a stationary dowel balance test by determining blood and brain ethanol concentrations at fall. Acute tolerance to the ataxic effects of ethanol was determined by measuring blood ethanol concentration (BEC) at regain of dowel balance ability after the first injection (BEC1RB) and after a second ethanol injection of 2.0 g/kg (BEC2RB). Acute tolerance was quantified by the difference in ethanol concentration at the two regains of balance (BEC2RB , BEC1RB) or by the difference between the second regain and one of the initial sensitivity measures (BEC2RB , initial sensitivity). Results: Four different measures of initial sensitivity were taken: two that used BEC values and two that used forebrain or hindbrain ethanol concentrations. We calculated acute tolerance values by using each of these initial sensitivity measures plus BEC2RB. No evidence of a genetic relationship between initial sensitivity and acute tolerance was found, which suggests that these are two independent phenomena with respect to stationary dowel balance. Conclusions: Three conclusions can be drawn from this work: (1) Orbital sinus BEC at early time points (<5 min postinjection) may or may not accurately reflect brain EC in mice, dependent on genotype; (2) there is no genetic relationship between initial sensitivity and acute tolerance to stationary dowel ataxia in the LSXSS RIs; and (3) sex-specific factors affect low-dose ethanol responses on the stationary dowel. [source] Imaging the effects of castration on bone turnover and hormone-independent prostate cancer colonization of boneTHE PROSTATE, Issue 15 2008N.A. Cross Abstract INTRODUCTION Tumor populations may selectively colonize bone that is being actively remodeled. In prostate cancer patients, androgen deprivation directly inhibits tumor growth initially, whilst induced bone loss may facilitate tumor colonization of bone by androgen-insensitive cells. We have tested this hypothesis using a xenograft model of early growth of prostate cancer in bone. METHODS PC3 cells transfected with Green fluorescent protein (GFP) were injected into castrated and non-castrated athymic mice via intrabial and intracardiac routes. In vivo tumor growth was monitored daily and animals sacrificed 6,9 days following initial GFP-based detection of tumors. Tumor bearing and contra-lateral non-tumor bearing tibias were analyzed extensively by micro-CT and histology/immunohistochemistry for the presence of tumor cells and the effects of tumor and/or castration on bone cells and bone structure evaluated. RESULTS GFP-positive tumors in bone were visible from 12 days post-injection following intratibial injection, allowing tumors <1 mm diameter to be monitored in live animals. Castration did not affect tumor frequency, tumor volume, or time to initial appearance of tumors injected via intratibial or intracardiac routes. Castration decreased trabecular bone volume in all mice. Significant tumor-induced suppression of numbers of osteoblasts, coupled with increased numbers of activated osteoclasts, was evident in both intact animals and castrated animals. CONCLUSIONS In vivo GFP imaging allows the detection of early tumor growth at intra-osseous sites. Castration induces bone loss, but PC3-GFP cells are also capable of inducing bone remodeling in intact animals at early time points, independently of pre-existing castration-induced alterations to bone. Prostate 68: 1707,1714, 2008. © 2008 Wiley-Liss, Inc. [source] Inflammatory stimuli accelerate Sjögren's syndrome,like disease in (NZB × NZW)F1 miceARTHRITIS & RHEUMATISM, Issue 5 2008Umesh S. Deshmukh Objective This study was undertaken to determine whether induction of systemic inflammation accelerates the development of Sjögren's syndrome (SS) in genetically susceptible mice. Methods Female (NZB × NZW)F1 mice were treated with either Freund's incomplete adjuvant (IFA) or phosphate buffered saline (PBS) at monthly intervals. Salivary gland function was monitored by measuring pilocarpine-induced saliva volume. Mice were killed at different time points and examined for sialadenitis and salivary gland,infiltrating cells. Sera were analyzed for autoantibodies to salivary gland antigens, nuclear antigens, and Ro60. Results While IFA-treated mice had significantly decreased salivary secretion 7 weeks after the initial treatment, salivary secretion did not decrease in PBS-treated controls until 17 weeks. At 7 weeks, the severity of sialadenitis and the number of T and B cells infiltrating the salivary glands did not differ between the 2 groups. However, at this time point IFA-treated mice showed significantly higher frequencies of CD11clow, B220+, Ly6C+, mouse PDCA-1+ dendritic cells (DCs) in the salivary glands. While levels of autoantibodies did not differ between the 2 groups at early time points, by late time points IFA-treated mice had higher levels. The gland dysfunction observed in IFA-treated mice at earlier time points did not correlate with the severity of sialadenitis or levels of autoantibodies. Instead, it was associated with increased frequency of plasmacytoid DCs in the gland. Conclusion Our data suggest that generalized inflammatory stimuli can accelerate the development of SS-like disease in (NZB × NZW)F1 mice, and that gland dysfunction in SS can develop prior to the generation of a robust adaptive autoimmune response. [source] Distinctive IGH gene segment usage and minimal residual disease detection in infant acute lymphoblastic leukaemiasBRITISH JOURNAL OF HAEMATOLOGY, Issue 2 2005Aihong Li Summary Infant acute lymphoblastic leukaemia (ALL) represents a rare but unique subset with poor prognosis. We analysed mixed-lineage leukaemia (MLL) gene rearrangements and the sequences of complete and incomplete immunoglobulin heavy chain gene rearrangements (IGH) in 14 infants (age ,12 months at diagnosis) enrolled on Dana-Farber Cancer Institute ALL Consortium Protocol 95,01. The dynamics of the leukaemic clone were followed during the course of the disease by quantitative real-time polymerase chain reaction of IGH rearrangements. Sixteen sequences were obtained from 13 (93%) of these infants. There was marked over usage of the VH6.1 gene segment (64%) in infants compared with older children with ALL (8%), (P < 0·001) and overusage of DH6 (P = 0·004) and JH1 (P = 0·004). Poor outcome was associated with MLL gene rearrangements rather than any specific VHDHJH gene usage patterns. Levels of minimal residual disease (MRD) at the end of induction appeared to be high in infants with ALL compared with older children, and although the number of infant cases studied was small, there were no differences in MRD levels after induction therapy in infant ALL with or without MLL gene rearrangements (P = 0·41) and quantitative MRD assessment at the early time points may not be predictive of outcome. Novel treatment strategies are required to improve the outcome in this poor prognosis subset of children with ALL. [source] Surfactant protein D deficiency influences allergic immune responsesCLINICAL & EXPERIMENTAL ALLERGY, Issue 12 2004B. Schaub Summary Background The collectin surfactant protein D (SP-D) confers protection against pulmonary infection and inflammation. Recent data suggest a role for SP-D in the modulation of allergic inflammation. Objective The aim of this study is to characterize the immune responses of SP-D-deficient (SP-D,/,) mice in a kinetic model of allergic inflammation. We determined whether allergic parameters were enhanced in SP-D,/, mice in vivo. Further, we examined whether functional immune responses in vitro such as lymphocyte proliferation (LP) and cytokine production were modulated in the absence of SP-D. Methods In vivo, wild-type (WT) and SP-D,/, mice were sensitized and challenged with the allergen ovalbumin (OVA) and assessed for allergic parameters (bronchoalveolar lavage (BAL) eosinophils, IL-13 production, pulmonary IFN-,, IL-10 expression) at early time points (1 and 3 days of challenge) in comparison with late time points (7 days of challenge). In vitro, spleen cells from WT and SP-D,/, mice were stimulated with the mitogen concanavalin A (ConA) and lipid A (LpA) and analysed for LP, IL-13 and IFN-, production. Toll-like receptor 4 (TLR4), ligand for LpA, was assessed by mRNA expression and immunohistochemistry in vivo. Results Following allergen exposure in vivo, SP-D,/, mice expressed higher BAL eosinophils and IL-13 concentrations and lower IFN-, expression at early time points compared with WT mice. IL-10 expression was increased at early time points in SP-D,/, compared with WT mice. Allergen-induced TLR4 expression was increased in WT, but not in SP-D,/, mice. After stimulation with LpA and ConA in vitro LP was increased and IFN-, concentration was decreased in SP-D,/, mice. Conclusion SP-D may be critical for the modulation of early stages of allergic inflammation in vivo. [source] | |||||||||||||