Low viscosity silicone with less shrinkage for brain slices.

Plastination consists of replacing lipid and water with a curable polymer. This technique has numerous advantages, of which the production of non-toxic, inert, highly durable, dry, and easy maintenance and storage specimens stand out. Like all anatomical techniques, plastination also has disadvantages, and one of them is tissue shrinkage. The feasibility of using low viscosity domestic silicone (0,1Pa.s at 20°C) to plastinate brain slices was examined. Twenty humans, 10 millimeters (mm) brain slices were impregnated, ten slices each with two polymers [10 with domestic low viscosity polymer - P1 and 10 slices with Biodur® (0,45-0,6Pa.s at 20°C) S10]. Shrinkage was accessed by volume and area measurements. Volume shrinkage was significantly less in the slices impregnated with low viscosity domestic polymer, demonstrating the feasibility to plastinate brain slices with domestic low viscosity silicone polymer.

Chromatographic method development using multivariate approaches for organic solvents optimized analysis in [18F]fluorocholine.

A fast and simple method using Gas Chromatography combined with Flame Ionization Detection (GC-FID) was developed for the determination of ethanol, acetonitrile, dibromomethane, dimethylaminoethanol, and dimethyl sulfoxide in [18F]fluorocholine. The combination of fractional factorial design, Doehlert design, and Desirability function was used to evaluate the operational parameters and to establish the best working condition. The validation results revealed that the proposed method has good recovery (85.1-104.1%) and repeatability (RSD ≤8.1%). Correlation coefficients (R ≥ 0.983) indicated good linearity over a wide range. The limit of detection (≤2.5 ppm) and the limit of quantification (≤7.5 ppm) were satisfactory. The proposed method is based on minimum manual operation, sample preparation free, direct injection technique, and short chromatographic separation time. This method is useful for routine analysis of organic solvents in [18F]fluorocholine, feasible for the modernization of specific monograph, and was therefore successfully implemented to assess samples manufactured by Nuclear Technology Development Center (CDTN).

A simple method for glass analysis using total reflection X-ray fluorescence spectrometry.

In this work, a simple and cost-effective method was developed for Na, Mg, Al, K, Ca, Ti, V, Cr, Mn, Ni, Cu, Zn, Rb, Sr, Ba and Pb determination in glass samples employing suspension sampling and total reflection X-ray fluorescence (TXRF). The experimental conditions were optimized using multivariate approach being the optimum conditions located by Derringer's desirability function. Proposed method was validated in accordance with the National Institute of Metrology, Quality and Technology (Inmetro, Brazil). Limit of detection (LD) and limit of quantification (LQ) were adequate for determination of trace elements in glass. The evaluation of the accuracy and precision was realized by analysis of standard reference materials of glass (NIST 612). For the majority of the elements, good agreement was achieved between the certified value and the value obtained in the NIST 612. The relative standard deviation (RSD%) was achieved between 3.6 and 10.3%. Moreover, no significant differences were observed between the proposed method compared to the ICP-MS. After the validation step, the method was applied to 31 glass samples and with aid of an exploratory principal component analysis (PCA), a perfect discrimination of the glass from originally manufactured smartphones was obtained. In addition, soda-lime glass could be reasonably distinguished from smartphone screens. The developed method is fast, consumes low amounts of reagents and allows the determination of many analytes simultaneously. Additionally, the method does not require calibration curves. Therefore, TXRF proved to be attractive and useful for routine analysis of glass.

Scoping Review of Oral Health-Related Birth Cohort Studies: Toward a Global Consortium.

The multidisciplinary nature and long duration of birth cohort studies allow investigation of the relationship between general and oral health and indicate the most appropriate stages in life to intervene. To date, the worldwide distribution of oral health-related birth cohort studies (OHRBCSs) has not been mapped, and a synthesis of information on methodological characteristics and outcomes is not available. We mapped published literature on OHRBCSs, describing their oral health-related data and methodological aspects. A 3-step search strategy was adopted to identify published studies using PubMed, Embase, Web of Science, and OVID databases. Studies with baseline data collection during pregnancy or within the first year of life or linked future oral health data to exposures during either of these 2 life stages were included. Studies examining only mothers' oral health and specific populations were excluded. In total, 1,721 articles were suitable for initial screening of titles and abstracts, and 528 articles were included in the review, identifying 120 unique OHRBCSs from 34 countries in all continents. The review comprised literature from the mid-1940s to the 21st century. Fifty-four percent of the OHRBCSs started from 2000 onward, and 75% of the cohorts were from high-income and only 2 from low-income countries. The participation rate between the baseline and the last oral health follow-up varied between 7% and 93%. Ten cohorts that included interventions were mostly from 2000 and with fewer than 1,000 participants. Seven data-linkage cohorts focused mostly on upstream characteristics and biological aspects. The most frequent clinical assessment was dental caries, widely presented as decayed, missing, and filled teeth (DMFT/dmft). Periodontal conditions were primarily applied as isolated outcomes or as part of a classification system. Socioeconomic classification, ethnicity, and country- or language-specific assessment tools varied across countries. Harmonizing definitions will allow combining data from different studies, adding considerable strength to data analyses; this will be facilitated by forming a global consortium.

Temporary Tooth Separation to Improve Assessment of Approximal Caries Lesions: A School-Based Study.

CLINICAL RELEVANCE: Radiographic lesion depth should not be used as the single determinant of the restorative threshold for clinically inaccessible approximal caries lesions. Temporary tooth separation is a feasible and effective diagnostic aid for assessment and appropriate management of approximal lesions. SUMMARY: In the era of tooth-preserving dentistry, the decision to restore approximal caries lesions must be based on the accurate assessment of tooth cavitation, as the accumulation of oral biofilms in these areas encourages lesion progression. However, lesions radiographically into dentin remain the main threshold criterion for restoring approximal lesions even though most of these lesions may not be cavitated. A school-based clinical protocol for temporary tooth separation (TTS) was developed to improve visual-tactile assessment and management of clinically inaccessible approximal lesions. TTS data retrieved from electronic health records were used to correlate radiographic lesion depth and surface cavitation status with lesion location and the patient's caries risk and to evaluate the effectiveness of TTS as a diagnostic aid for approximal lesions. Of the 206 lesions assessed, 66.5% (n=137) were located in the maxillary arch, 56.6% (n=116) in distal surfaces, 61.3% (n=114) in premolars, and 21.5% (n=40) in molars. After tooth separation, 79.6% (n=164) of the lesions were diagnosed as noncavitated, including 90% (n=66) of the lesions radiographically at the inner half of enamel (E2) and 66% (n=49) of those at the outer-third of dentin (D1). Logistic regression analysis using E2 and D1 lesions showed no significant association between lesion depth or cavitation status with lesion location and caries risk. TTS is a feasible and effective diagnostic aid for the assessment and appropriate management of approximal caries lesions. There is a need to reevaluate the use of radiographic lesion depth as the single determinant of the restorative threshold for clinically inaccessible approximal lesions.

Protective role of jaboticaba Plinia peruviana peel extract in copper-induced cytotoxicity in Allium cepa.

Jaboticaba Plinia peruviana (Poir.) Govaerts is a Brazilian berry that presents high levels of polyphenols, which may play a key role in preventing cytotoxic and genotoxic effects of harmful agents. Although copper is an essential micronutrient that plays an important role in organisms, high copper concentrations may trigger toxicity to animals and plants. Here, we investigated whether Plinia peruviana hydroalcoholic extract prevents copper-induced cytotoxicity in Allium cepa root cells. Five different anthocyanins and phenolic compounds were identified in Plinia peruviana extract. Importantly, the exposure to 1.53 mg/L copper for 24 h impaired mitotic index, as well as increased mitosis disturbances and triggered DNA damage. Pre-incubation with Plinia peruviana extract (0.25 g/L and 0.75 g/L) for 3 h prevented copper-induced changes in the mitotic index and reduced the number of abnormal cells. In conclusion, we suggest that Plinia peruviana peel extract has protective effects against cellular and genetic disturbances induced by copper.

Protective effect of Uncaria tomentosa extract against oxidative stress and genotoxicity induced by glyphosate-Roundup® using zebrafish (Danio rerio) as a model.

Oxidative stress and DNA damage are involved in the glyphosate-based herbicide toxicity. Uncaria tomentosa (UT; Rubiaceae) is a plant species from South America containing bioactive compounds with known beneficial properties. The objective of this work was to evaluate the antioxidant and antigenotoxic potential of UT extract in a model of acute exposure to glyphosate-Roundup® (GR) in zebrafish (Danio rerio). We showed that UT (1.0 mg/mL) prevented the decrease of brain total thiols, the increase of lipid peroxidation in both brain and liver, and the decrease of liver GPx activity caused after 96 h of GR (5.0 mg/L) exposure. In addition, UT partially protected against the increase of micronucleus frequency induced by GR exposure in fish brain. Overall, our results indicate that UT protects against damage induced by a glyphosate-based herbicide by providing antioxidant and antigenotoxic effects, which may be related to the phenolic compounds identified in the extract.

Gender differences in aggression and cortisol levels in zebrafish subjected to unpredictable chronic stress.

Chronic stress may cause physical, behavioral and neuropsychiatric changes, affecting the health condition of an individual. Aggression is a universal behavior with great relevance on human and animal social systems. Despite studies showing the influence of chronic stress on aggression, the effects of unpredictable chronic stress (UCS) on aggressive behavior in male and female zebrafish remain unknown. Thus, the aim of this study was to evaluate the effects of UCS on the aggressive behavior and cortisol levels in adult zebrafish of both sexes. Our results showed that UCS increased aggression in males, but not in females, which displayed more aggressive behavior at baseline than control males. Increased whole-body cortisol levels were observed in stressed males; however, no differences were found between female groups. In conclusion, we reported for the first time gender differences on behavioral parameters and cortisol levels in response to UCS in zebrafish. These results highlight the relevance of studying behavioral and physiological parameters in both sexes separately.

Prevention of unpredictable chronic stress-related phenomena in zebrafish exposed to bromazepam, fluoxetine and nortriptyline.

RATIONALE: Several model organisms have been employed to study the impacts of stress on biological systems. Different models of unpredictable chronic stress (UCS) have been established in rodents; however, these protocols are expensive, long-lasting, and require a large physical structure. Our group has recently reported an UCS protocol in zebrafish with several advantages compared to rodent models. We observed that UCS induced behavioral, biochemical, and molecular changes similar to those observed in depressed patients, supporting the translational relevance of the protocol. OBJECTIVES: Considering that a pharmacological assessment is lacking in this zebrafish model, our aim was to evaluate the effects of anxiolytic (bromazepam) and antidepressant drugs (fluoxetine and nortriptyline) on behavioral (novel tank test), biochemical (whole-body cortisol), and molecular parameters (cox-2, tnf-α, il-6, and il-10 gene expression) in zebrafish subjected to UCS. RESULTS: We replicated previous data showing that UCS induces behavioral and neuroendocrine alterations in zebrafish, and we show for the first time that anxiolytic and antidepressant drugs are able to prevent such effects. Furthermore, we extended the molecular characterization of the model, revealing that UCS increases expression of the pro-inflammatory markers cox-2 and il-6, which was also prevented by the drugs tested. CONCLUSIONS: This study reinforces the use of zebrafish as a model organism to study the behavioral and physiological effects of stress. The UCS protocol may also serve as a screening tool for evaluating new drugs that can be used to treat psychiatric disorders with stress-related etiologies.

Effect of xylitol varnishes on remineralization of artificial enamel caries lesions in situ.

OBJECTIVES: Analyze the effect of varnishes containing xylitol compared to commercial fluoridated varnishes on the remineralization of artificial enamel caries lesions in situ. METHODS: Twenty subjects took part in this crossover, double-blind study performed in four phases of 5days each. Each subject worn palatal appliances containing four predemineralized bovine enamel specimens. Artificial caries lesions were produced by immersion in 30ml of lactic acid buffer containing 3mM CaCl2·2H2O, 3mM KH2PO4, 6µM tetraetil metil diphosphanate (pH 5.0) for 6days. The specimens in each subject were treated once with the following varnishes: 20% xylitol (experimental); Duofluorid™ (6% NaF, 6% CaF2), Duraphat™ (5% NaF, positive control) and placebo (no-F/xylitol, negative control). The varnishes were applied in a thin layer and removed after 6h. Fifteen subjects were able to finish all phases. The enamel alterations were quantified by surface hardness and transversal microradiography. The percentage of surface hardness recovery (%SHR), the integrated mineral loss and lesion depth were statistically analyzed by Friedmann and Dunn's tests test (p<0.05). RESULTS: Enamel surface remineralization was significantly increased by Duraphat™, Duofluorid™ and 20% xylitol formulations. Significant subsurface mineral remineralization could also be seen for the experimental and commercial varnishes, except for Duraphat™, when the parameter "lesion depth" was considered. CONCLUSIONS: 20% xylitol varnish seem to be a promising alternative to increase surface and subsurface remineralization of artificial caries lesions in situ. CLINICAL SIGNIFICANCE: effective vehicles are desirable for caries control. Xylitol varnishes seem to be promising alternatives to increase enamel remineralization in situ, which should be confirmed by clinical studies.

Fluoxetine and diazepam acutely modulate stress induced-behavior.

Drug residue contamination in aquatic ecosystems has been studied extensively, but the behavioral effects exerted by the presence of these drugs are not well known. Here, we investigated the effects of acute stress on anxiety, memory, social interaction, and aggressiveness in zebrafish exposed to fluoxetine and diazepam at concentrations that disrupt the hypothalamic-pituitary-interrenal (HPI) axis. Stress increased the locomotor activity and time spent in the bottom area of the tank (novel tank). Fluoxetine and diazepam prevented these behaviors. We also observed that stress and fluoxetine and diazepam exposures decreased social interaction. Stress also increased aggressive behavior, which was not reversed by fluoxetine or diazepam. These data suggest that the presence of these drugs in aquatic ecosystems causes significant behavioral alterations in fish.

N-acetylcysteine prevents stress-induced anxiety behavior in zebrafish.

Despite the recent advances in understanding the pathophysiology of anxiety disorders, the pharmacological treatments currently available are limited in efficacy and induce serious side effects. A possible strategy to achieve clinical benefits is drug repurposing, i.e., discovery of novel applications for old drugs, bringing new treatment options to the market and to the patients who need them. N-acetylcysteine (NAC), a commonly used mucolytic and paracetamol antidote, has emerged as a promising molecule for the treatment of several neuropsychiatric disorders. The mechanism of action of this drug is complex, and involves modulation of antioxidant, inflammatory, neurotrophic and glutamate pathways. Here we evaluated the effects of NAC on behavioral parameters relevant to anxiety in zebrafish. NAC did not alter behavioral parameters in the novel tank test, prevented the anxiety-like behaviors induced by an acute stressor (net chasing), and increased the time zebrafish spent in the lit side in the light/dark test. These data may indicate that NAC presents an anti-stress effect, with the potential to prevent stress-induced psychiatric disorders such as anxiety and depression. The considerable homology between mammalian and zebrafish genomes invests the current data with translational validity for the further clinical trials needed to substantiate the use of NAC in anxiety disorders.

Depressive symptoms and alcohol correlates among Brazilians aged 14 years and older: a cross-sectional study.

BACKGROUND: The associations between depressive symptoms and alcohol-related disorders, drinking patterns and other characteristics of alcohol use are important public health issues worldwide. This study aims to study these associations in an upper middle-income country, Brazil, and search for related socio-demographic correlations in men and women. METHODS: A cross-sectional study was conducted between November 2005 and April 2006. The sample of 3,007 participants, selected using a multistage probabilistic sampling method, represents the Brazilian population aged 14 and older. Depressive symptoms were assessed using the Center for Epidemiologic Studies Depression Scale and alcohol dependence was assessed using the Composite International Diagnostic Interview. Associations assessed using bi-variate analysis were tested using Rao-Scott measures. Gender specific multinomial logistic regression models were developed. RESULTS: Among the participants with alcohol dependence, 46% had depressive symptoms (17.2% mild/moderate and 28.8% major/severe; p < 0.01); 35.8% (p = 0.08) of those with alcohol abuse and 23.9% (p < 0.01) of those with a binge-drinking pattern also had depressive symptoms. Alcohol abstainers and infrequent drinkers had the highest prevalence of major/severe depressive symptoms, whereas frequent heavy drinkers had the lowest prevalence of major/severe depressive symptoms. In women, alcohol dependence and the presence of one or more problems related to alcohol consumption were associated with higher risks of major/severe depressive symptoms. Among men, alcohol dependence and being ≥45 years old were associated with higher risks of major/severe depressive symptoms. CONCLUSIONS: In Brazil, the prevalence of depressive symptoms is strongly related to alcohol dependence; the strongest association was between major/severe depressive symptoms and alcohol dependence in women. This survey supports the possible association of biopsychosocial distress, alcohol consumption and the prevalence of depressive symptoms in Brazil. Investing in education, social programs, and care for those with alcohol dependence and major/severe depressive symptoms, especially for such women, and the development of alcohol prevention policies may be components of a strategic plan to reduce the prevalence of depression and alcohol problems in Brazil. Such a plan may also promote the socio-economic development of Brazil and other middle-income countries.

Subchronic atrazine exposure changes defensive behaviour profile and disrupts brain acetylcholinesterase activity of zebrafish.

Animal behaviour is the interaction between environment and an individual organism, which also can be influenced by its neighbours. Variations in environmental conditions, as those caused by contaminants, may lead to neurochemical impairments altering the pattern of the behavioural repertoire of the species. Atrazine (ATZ) is an herbicide widely used in agriculture that is frequently detected in surface water, affecting non-target species. The zebrafish is a valuable model organism to assess behavioural and neurochemical effects of different contaminants since it presents a robust behavioural repertoire and also all major neurotransmitter systems described for mammalian species. The goal of this study was to evaluate the effects of subchronic ATZ exposure in defensive behaviours of zebrafish (shoaling, thigmotaxis, and depth preference) using the split depth tank. Furthermore, to investigate a putative role of cholinergic signalling on ATZ-mediated effects, we tested whether this herbicide alters acetylcholinesterase (AChE) activity in brain and muscle preparations. Fish were exposed to ATZ for 14days and the following groups were tested: control (0.2% acetone) and ATZ (10 and 1000µg/L). The behaviour of four animals in the same tank was recorded for 6min and biological samples were prepared. Our results showed that 1000µg/L ATZ significantly increased the inter-fish distance, as well as the nearest and farthest neighbour distances. This group also presented an increase in the shoal area with decreased social interaction. No significant differences were detected for the number of animals in the shallow area, latency to enter the shallow and time spent in shallow and deep areas of the apparatus, but the ATZ 1000 group spent significantly more time near the walls. Although ATZ did not affect muscular AChE, it significantly reduced AChE activity in brain. Exposure to 10µg/L ATZ did not affect behaviour or AChE activity. These data suggest that ATZ impairs defensive behaviours of zebrafish, which could be related to its action on brain cholinergic neurotransmission. Moreover, the use of the split depth tank could be an alternative strategy to assess group behaviour and depth preference after exposure to chemical compounds.

Inhibition of tooth erosion by milk containing different fluoride concentrations: an in vitro study.

OBJECTIVES: This in vitro study assessed the effect of milk containing different fluoride concentrations on tooth erosion. METHODS: Bovine enamel and root dentine specimens were treated with: (1) bovine whole milk with 0ppm F; (2) 2.5ppm F; (3) 5ppm F; (4) 10ppm F (all after erosion); (5) whole milk with 0ppm F (before erosion); (6) NaF (0.05% F, positive control, after erosion) or (7) 0.9% NaCl (negative control, after erosion). The specimens were submitted to pH cycles (4× 90s in soft drink) and treatments for 5 days. The specimens were immersed in the treatment solutions for 1min (only at the first cycle each day) with further exposition to 1:1 milk:saliva slurry for 10min. The tooth loss was measured using a contact profilometer and statistically analysed (p<0.05). RESULTS: Rinsing with milk before erosive challenge significantly reduced tooth loss compared to negative control (67% and 24% reduction in dentine and enamel loss, respectively) and to milk after erosive challenge, only for dentine. The addition of fluoride to milk also reduced tooth loss compared to negative control, but with no significant differences among fluoride concentrations for enamel and dentine (µm), respectively: 0ppm (3.63±0.04 and 2.51±0.53), 2.5ppm F (2.86±0.42 and 1.96±0.47), 5ppm F (2.81±0.27 and 1.77±0.44), 10ppm F (2.03±0.49 and 1.68±0.59). There was a negative and significant correlation between [F] and the tooth loss. CONCLUSIONS: Daily rinse with milk containing F is able to reduce both enamel and dentine erosion in vitro. CLINICAL SIGNIFICANCE: Since the prevalence of dental erosion is steadily increasing, rinse with milk or its derivate might be an important strategy to reduce the progression of tooth erosion.

Higher prevalence of major depressive symptoms in Brazilians aged 14 and older.

OBJECTIVE: Depression is a highly prevalent condition and is considered a major public health issue. The aim of the present study was to estimate the prevalence of depressive symptoms in the Brazilian population and establish their sociodemographic correlates. METHOD: A cross-sectional study was conducted between November 2005 and April 2006. Data were collected in face-to-face interviews using a standardized questionnaire. The sample consisted of 3,007 interviews with individuals aged 14 years and older and followed a probabilistic design covering the Brazilian national territory. Depressive symptoms were assessed according to the Center for Epidemiologic Studies Depression Scale. RESULTS: The observed prevalence of depressive symptoms was 28.3% (13% mild/moderate; 15.3% major/severe; p < 0.01). Increased depressive symptom rates were associated with being a female, being 45 years of age and older, having lower educational attainment, being single, having family income of up to 2.5 times minimum wage, and living in the northern region of Brazil (p < 0.05). CONCLUSIONS: The prevalence of depressive symptoms in Brazil is high, with major depressive symptoms being the most frequent form of this symptomatology. Considering the biopsychosocial model of mental disorders, this survey points to the involvement of psychosocial factors in the prevalence of depressive symptoms in Brazil.

Higher prevalence of major depressive symptoms in Brazilians aged 14 and older

Objective: Depression is a highly prevalent condition and is considered a major public health issue. The aim of the present study was to estimate the prevalence of depressive symptoms in the Brazilian population and establish their sociodemographic correlates. Method: A cross-sectional study was conducted between November 2005 and April 2006. Data were collected in face-to-face interviews using a standardized questionnaire. The sample consisted of 3,007 interviews with individuals aged 14 years and older and followed a probabilistic design covering the Brazilian national territory. Depressive symptoms were assessed according to the Center for Epidemiologic Studies Depression Scale. Results: The observed prevalence of depressive symptoms was 28.3% (13% mild/moderate; 15.3% major/severe; p < 0.01). Increased depressive symptom rates were associated with being a female, being 45 years of age and older, having lower educational attainment, being single, having family income of up to 2.5 times minimum wage, and living in the northern region of Brazil (p < 0.05). Conclusions: The prevalence of depressive symptoms in Brazil is high, with major depressive symptoms being the most frequent form of this symptomatology. Considering the biopsychosocial model of mental disorders, this survey points to the involvement of psychosocial factors in the prevalence of depressive symptoms in Brazil. .

A novel TNF receptor family member binds TWEAK and is implicated in angiogenesis.

TWEAK is a member of the TNF ligand family that induces angiogenesis in vivo. We report cloning of a receptor for TWEAK (TweakR) from a human umbilical vein endothelial cell (HUVEC) library. The mature form of TweakR has only one hundred and two amino acids and six cysteine residues in its extracellular region. Five different assays demonstrate TWEAK-TweakR binding, and the interaction affinity constant (Kd) is within a physiologically relevant range of 2.3 +/- 0.1 nM. The TweakR cytoplasmic domain binds TRAFs 1, 2, and 3. Cross-linking of TweakR induces HUVEC growth, and mRNA levels are upregulated in vitro by a variety of agents and in vivo following arterial injury. Soluble TweakR inhibits endothelial cell migration in vitro and corneal angiogenesis in vivo.

ULBP1, 2, 3: novel MHC class I-related molecules that bind to human cytomegalovirus glycoprotein UL16, activate NK cells.

New members of the extended MHC class I-like family were identified based on their ability to bind human cytomegalovirus glycoprotein UL16 and/or their mutual homology. Soluble UL16 binding proteins (ULBP) competed with each other for binding to NK cells. Treatment of human and mouse NK cells with ULBP led to increased production of cytokines/chemokines, proliferation, cytotoxic activity and up-regulation of activation-associated surface molecules. The presence of ULBP during the stimulation phase of the CTL assay caused increased cytotoxic activity. Addition of soluble recombinant UL16 protein inhibited the biological activities mediated by ULBP, suggesting the existence of a novel mechanism utilized by CMV to evade elimination by the host immune system.

CD14 employs hydrophilic regions to "capture" lipopolysaccharides.

CD14 participates in the host innate inflammatory response to bacterial LPS obtained from Escherichia coli and other Gram-negative bacteria. Evidence from several laboratories suggests that different regions of the amino-terminal portion of the molecule may be involved in LPS binding. In this report a series of single-residue serine replacement and charge reversal mutations were generated to further elucidate the mechanism by which this protein may bind a multitude of different LPS ligands. Single-residue CD14 mutation proteins were examined for their ability to bind LPS obtained from E. coli, Porphyromonas gingivalis, and Helicobacter pylori and facilitate the activation of E-selectin from human endothelial cells. In addition, the single-residue CD14 mutation proteins were employed to perform monoclonal epitope-mapping studies with three LPS-blocking Abs that bound tertiary epitopes. Evidence that several different hydrophilic regions of the amino-terminal region of CD14 are involved in LPS binding was obtained. Epitope-mapping studies revealed that these hydrophilic regions are located on one side of the protein surface. These studies suggest that CD14 employs a charged surface in a manor similar to the macrophage scavenger receptor to "capture" LPS ligands and "present" them to other components of the innate host defense system.