Environmentally Friendly New Catalyst Using Waste Alkaline Solution from Aluminum Production for the Synthesis of Biodiesel in Aqueous Medium.

Red mud (RM) is composed of a waste alkaline solution (pH = 13.3) obtained from the production of alumina. It contains high concentrations of hematite (Fe2O3), goethite (FeOOH), gibbsite [Al(OH)3], a boehmite (AlOOH), anatase (Tetragonal-TiO2), rutile (Ditetragonal dipyramidal-TiO2), hydrogarnets [Ca3Al2(SiO4)3-x(OH)4x], quartz (SiO2), and perovskite (CaTiO3). It was shown to be an excellent catalytic mixture for biodiesel production. To demonstrate the value of RM, an environmentally friendly process of transesterification in aqueous medium using waste cooking oil (WCO), MeOH, and waste alkaline solution (WAS) obtained from aluminum production was proposed. Triglycerides of WCO reacted with MeOH at 60 °C to yield mixtures of fatty acid methyl esters (FAMEs) in the presence of 0.019% (w/w) WAS/WCO using the WAS (0.204 mol L-1, predetermined by potentiometric titration) from aluminum production by the Bayer process. The use of the new catalyst (WAS) resulted in a high yield of the products (greater than 99% yield).

Influence of dietary total antioxidant capacity on the association between smoking and hypertension in Brazilian graduates (CUME project).

BACKGROUND AND AIMS: Hypertension (HTN) is a chronic non-communicable disease influenced by non-modifiable risk factors, such as sex and age, as well as modifiable risk factors such as lifestyle, including diet and smoking. Moreover, diet quality among smokers is worse than that of non-smokers, mainly in terms of antioxidant content. Thus, the current study aimed to investigate whether dietary total antioxidant capacity (dTAC) influences the association between smoking and HTN. METHODS AND RESULTS: This cross-sectional study included 4303 graduates (69.35% women) from the Cohort of Minas Gerais Universities (CUME) project. An online food frequency questionnaire was administered to participants, and dTAC was estimated using the ferric reducing antioxidant power method. In the questionnaires, individuals reported smoking status, systolic and diastolic blood pressure values, previous HTN diagnosis, and use of antihypertensive drugs. Logistic regression models were used to estimate the odds ratio and 95% confidence interval between smoking and HTN, stratified by the median dTAC. Current and former smokers had higher dTAC values despite their lower fruit intake. Moreover, coffee was the main contributor to dTAC among them. Smoking was associated with a higher likelihood of HTN, mainly among individuals with a higher dTAC. However, after exclusion of coffee antioxidant capacity, there was an association between only smoking and HTN in individuals with lower dTAC. CONCLUSIONS: The controversial association between higher dTAC and HTN can result from high coffee intake. Higher dTAC without coffee intake may mitigate the association between smoking and HTN in this population.

Microwave-assisted catalytic fast pyrolysis of rice husk over a hierarchical HZSM-5/MCM-41 catalyst prepared by organic base alkaline solutions.

This paper reports the results obtained for microwave-assisted catalytic fast pyrolysis (MACFP) of rice husk. The MACFP process employed a hierarchical catalyst prepared via a combination of organic alkali treatment (TPAOH) and the generation of an external layer of MCM-41-type mesoporous channels. We propose this catalyst which is used for the first time for pyrolysis of lignocellulosic biomass, as a tool to reduce coke agglomeration and increase hydrocarbon yields. Our results indicate that during catalyst preparation the mass fraction of cetyltrimethylammonium bromide (CTAB) has a direct effect on the content of MCM-41 formed on top of the HZSM-5 core. For MACFP, we hypothesize that the small molecules generated from thermal decomposition of rice husk react further to form aromatic and aliphatic hydrocarbons by decarbonylation, decarboxylation, oligomerization and aromatization. The highest hydrocarbon yield (60.5%) was obtained for a catalyst modified by a 2.0 mol/L TPAOH solution, with 10 wt% of CTAB (template for producing MCM-41), as well as with digestion and crystallization at 110 °C for 24 h. In addition, the highest liquid yield (47.6 wt%) was obtained at 550 °C. The relative content of hydrocarbons goes through a maximum of 60.5% with CTAB mass fraction which was higher than values obtained with MCM-41 (3.2%) and HZSM-5 (36.0%). Characterization and catalytic testing results suggest that the digestion temperature plays a more important role in the catalyst synthesis than the crystallization temperature. High digestion temperature (120 °C) decreases the overall hydrocarbon selectivity from 60.5% (110 °C) to 39.2%. The relative content of oxygenates reached the lowest value of 35.9% at the digestion and crystallization temperature of 110 °C. The synergistic effect of the MCM-41 shell and the HZSM-5 core promotes the catalytic activity, leading to outstanding deoxygenation capabilities and excellent selectivity to BTEX (52.7%).

Maternal Passive Immunity and Dengue Hemorrhagic Fever in Infants.

Dengue hemorrhagic fever (DHF) can occur in primary dengue virus infection of infants [Formula: see text] year of age. To understand the presumed role of maternal dengue-specific antibodies received until birth in the development of this primary DHF in infants, we investigated a mathematical model based on a system of nonlinear ordinary differential equations that mimics cells, virus and antibodies interactions. The neutralization and enhancement activities of maternal antibodies against the virus are represented by a function derived from experimental data and knowledge from the medical literature. The analytic study of the model shows the existence of two equilibriums, a disease-free equilibrium and an endemic one. We performed the asymptotic stability analysis for these two equilibriums. The local asymptotic stability of the endemic equilibrium (DHF equilibrium) corresponds to the occurrence of DHF. Numerical results are also presented in order to illustrate the mathematical analysis performed, highlighting the most important parameters that drive model dynamics. We defined the age at which DHF occurs as the time when the infection takes off that means at the inflection point of the curve of infected cell population. We showed that this age corresponds to the one at which maximum enhancing activity for dengue infection appears. This critical time for the occurrence of DHF is calculated from the model to be approximately 2 months after the time for maternal dengue neutralizing antibodies to degrade below a protective level, which corresponds to what is observed in the experimental data from the literature.

Vanishing bile duct syndrome related to DILI and Hodgkin lymphoma overlap: A rare and severe case.

Vanishing bile duct syndrome is a rare acquired condition, characterized by progressive loss of intrahepatic bile ducts leading to ductopenia and cholestasis. It can be associated with infections, ischemia, drug adverse reactions, neoplasms, autoimmune disease, and allograft rejection. Prognosis is variable and depends on the etiology of bile duct injury. We report the case of a 25-year-old female with cholestatic hepatitis and concomitant intakes of hepatotoxic substances, such as garcinia, field horsetail, and ketoprofen. On suspicion of a drug-induced liver injury, the drugs were promptly withdrawn and ursodeoxycholic acid was started with initial clinical and laboratory improvement, and the patient was discharged from the hospital. One month later, she had a new increase in bilirubin levels and canalicular enzymes, requiring a liver biopsy that showed significant loss of intrahepatic bile ducts, which was compatible with vanishing bile duct syndrome. This was confirmed by using cytokeratin 19 on immunohistochemistry. There was subsequent lymph node enlargement in several chains, and relevant weight loss. Histological analysis of a cervical lymph node revealed nodular sclerosis-subtype classic Hodgkin lymphoma. In this setting, vanishing bile duct syndrome was related to Hodgkin lymphoma and a drug-induced liver injury overlap, leading to progressive cholestasis with a worse prognosis. The patient's response to chemotherapy was poor, requiring biological therapy with brentuximab vedotin. It is crucial for physicians to create a broad differential diagnosis in suspected vanishing bile duct syndrome patients, especially to rule out malignancies.

Effect of alkali-treated HZSM-5 zeolite on the production of aromatic hydrocarbons from microwave assisted catalytic fast pyrolysis (MACFP) of rice husk.

We performed microwave-assisted catalytic fast pyrolysis (MACFP) of rice husk (RH) over an alkali-treated HZSM-5 zeolite, for production of hydrocarbons. The treatment consisted in the modification of the HZSM-5 by the organic base tetrapropylammonium hydroxide (TPAOH) solution at several concentrations. We characterized the resulting catalysts by powder X-ray diffraction (XRD), transmission electron microscopy (TEM), N2 adsorption-desorption, and temperature-programmed sorption of ammonia (NH3-TPD). The results suggest that the TPAOH treatment generated mesoporous structures in the HZSM-5, while preserving its microporous structure and crystallinity. We obtained the highest yield (45.9%) of hydrocarbons from MACFP of rice husk (RH) at 550 °C. As the TPAOH concentration increases, the relative content of BTEX hydrocarbons (benzene, toluene, ethylbenzene, and xylene) reaches a maximum value of 22.9% at 2.0 mol/L. A comparison of results obtained over the organic base TPAOH (HZSM-5 modified by 2.0 mol/L TPAOH solution) with those obtained over an inorganic base (HZSM-5 modified by 2.0 mol/L NaOH solution) shows a 4.3% increase in the relative content of monocyclic aromatic hydrocarbons for the TPAOH. In addition, the TPAOH-treated catalyst shows excellent selectivity of BTEX (58.5%), which is higher than the selectivity obtained with the parent HZSM-5 (51.2%) and NaOH-treated HZSM-5 (53.9%). The TPAOH-modified HZSM-5 catalyst effectively reduced coke formation by 4.6% compared to MACFP over the parent HZSM-5, most likely because TPAOH decreases the concentration of strong acidic sites on the outer surface of the catalyst, creating a mesoporous structure while retaining the weak acidic sites on the HZSM-5 inner surface. The new catalyst generated in this work contains a moderate amount of mesopores structures, which allows for effective upgrading of pyrolysis vapors while simultaneously reducing coke formation, thereby addressing a significant problem in the development of the catalytic fast pyrolysis process.

Catalytic fast co-pyrolysis of waste greenhouse plastic films and rice husk using hierarchical micro-mesoporous composite molecular sieve.

Catalytic fast co-pyrolysis of waste greenhouse plastic films and rice husk over a hierarchical HZSM-5/MCM-41 catalyst was performed in an analytical Py-GC/MS. We evaluated the effect of pyrolysis temperature and the ratio of rice husk to waste greenhouse plastic films on the total peak area of condensable organic products and CO2. In order to evaluate synergy possibilities among the two feedstocks, we performed non-catalytic pyrolysis and catalytic fast pyrolysis of rice husk and waste greenhouse plastic films separately. In addition, we report results for the catalytic fast co-pyrolysis of the mixture rice husk and waste greenhouse plastic films. The maximum relative content of hydrocarbons from catalytic fast co-pyrolysis of rice husk and waste greenhouse plastic films is obtained at 600 °C. When the mass ratio of rice husk to waste greenhouse plastic films is 1:1.5, the relative content of hydrocarbons reaches a maximum (71.1%). The hierarchical micro-mesoporous composite molecular sieve used in this work has outstanding catalytic activity and increases the relative content of hydrocarbons.

Border analysis for spatial clusters.

BACKGROUND: The spatial scan statistic is widely used by public health professionals in the detection of spatial clusters in inhomogeneous point process. The most popular version of the spatial scan statistic uses a circular-shaped scanning window. Several other variants, using other parametric or non-parametric shapes, are also available. However, none of them offer information about the uncertainty on the borders of the detected clusters. METHOD: We propose a new method to evaluate uncertainty on the boundaries of spatial clusters identified through the spatial scan statistic for Poisson data. For each spatial data location i, a function F(i) is calculated. While not a probability, this function takes values in the [0, 1] interval, with a higher value indicating more evidence that the location belongs to the true cluster. RESULTS: Through a set of simulation studies, we show that the F function provides a way to define, measure and visualize the certainty or uncertainty of each specific location belonging to the true cluster. The method can be applied whether there are one or multiple detected clusters on the map. We illustrate the new method on a data set concerning Chagas disease in Minas Gerais, Brazil. CONCLUSIONS: The higher the intensity given to an area, the higher the plausibility of that particular area to belong to the true cluster in case it exists. This way, the F function provides information from which the public health practitioner can perform a border analysis of the detected spatial scan statistic clusters. We have implemented and illustrated the border analysis F function in the context of the circular spatial scan statistic for spatially aggregated Poisson data. The definition is clearly independent of both the shape of the scanning window and the probability model under which the data is generated. To make the new method widely available to users, it has been implemented in the freely available SaTScan[Formula: see text] software www.satscan.org .

Anti-helminthic activity of Momordica charantia L. against Fasciola hepatica eggs after twelve days of incubation in vitro.

Fasciolosis, a parasitic disease caused by the trematode Fasciola hepatica underreported is expanding both in human and animal population, throughout the world. The constant use of synthetic drugs to treat this condition has led to the natural selection of resistant strains of the parasite. Hence, there is a growing focus on the potential anti-helminthic properties of medicinal plants and phytopharmaceuticals. The current study assessed the potential anti-fasciolicide action of Momordica charantia leaf extracts and fractions on the eggs of F. hepatica parasites. The lyophilized crude extract (CE) of M. charantia leaves and its sub-fractions, obtained from liquid-liquid partitioning with organic solvents, were analysed by High Performance Liquid Chromatography (HPLC), suspended in 1% DMSO and used in in vitro tests. Quadruplicates of 50F. hepatica eggs were incubated at 23°C with M. charantia leaf CE in different concentrations. After 12days no larvae were formed in eggs incubated with CE concentrations above 12.5mg/mL. Eggs incubated with CE sub-fractions at concentrations of 1000, 100, 10, 1, 0.1, 0.01µg/mL affected embryonic development, with n-butanol presenting the strongest inhibition of miracidia formation. In contrast, on the 12th day, 90% of the miracidia hatched in the control experiments using 0.03% DMSO whereas embryogenesis was completely abolished with any concentration of albendazole sulphoxide ABZ(SO). Chemical analysis of the CE and sub-fractions revealed a prominent presence of flavonoids. HPLC-MS confirmed Quercetin to be one of the main flavonoids present in the CE and the n-butanol subfraction. This is the first study to analyse the potential anti-fasciolicide action of M. charantia leaf CE and subfractions.

Evaluation of the composition of Carica papaya L. seed oil extracted with supercritical CO2.

Among the most important tropical fruit grown in the world today and in Brazil, papaya occupies a prominent place. Native to tropical America, papaya has spread to several regions of the world, and Brazil accounts for 12.74% of the world production, followed by Mexico, Nigeria and India. The culture reached a harvested area of 441,042 ha and production of 12,420,585 t worldwide. The largest interest in this fruit relies on its main constituent compounds, like vitamins A, B and C, alkaloids (carpaine and pseudocarpaine), proteolytic enzymes (papain and quimiopapain) and benzyl isothiocyanate, more known as BITC, which has anthelmintic activity. Because of that, the present work has as objective the evaluation of the efficiency and composition of the oil extracted from Carica papaya L. seeds with supercritical carbon dioxide. The experiments were performed in a unit containing mainly a high-pressure pump and a stainless steel extractor with 42 mL of volume. The sampling was performed at each 20 min until the saturation of the process. About 6.5 g of sample were fed for each experiment done at 40, 60 and 80 °C under the pressures of 100, 150 and 200 bar. Samples of the Carica papaya L. fruit were acquired in a popular market and free for personal use intended for the study. After collection, the seeds were crushed with the help of a pestle, and dried at 60 °C for 60 min. For each operational condition, the extraction curves were constructed relating cumulative mass of oil extracted in function of the operational time. The better efficiencies were found at 40 °C and 200 bar (1.33%) followed by 80 °C and 200 bar (2.56%). Gas chromatography and NMR analysis could identify an insecticide component (BITC) that enables new applications of this residue in pharmaceutical and chemical industries.

Multiobjective Genetic Algorithm applied to dengue control.

Dengue fever is an infectious disease caused by a virus of the Flaviridae family and transmitted to the person by a mosquito of the genus Aedes aegypti. This disease has been a global public health problem because a single mosquito can infect up to 300 people and between 50 and 100 million people are infected annually on all continents. Thus, dengue fever is currently a subject of research, whether in the search for vaccines and treatments for the disease or efficient and economical forms of mosquito control. The current study aims to study techniques of multiobjective optimization to assist in solving problems involving the control of the mosquito that transmits dengue fever. The population dynamics of the mosquito is studied in order to understand the epidemic phenomenon and suggest strategies of multiobjective programming for mosquito control. A Multiobjective Genetic Algorithm (MGA_DENGUE) is proposed to solve the optimization model treated here and we discuss the computational results obtained from the application of this technique.

Data-driven inference for the spatial scan statistic.

BACKGROUND: Kulldorff's spatial scan statistic for aggregated area maps searches for clusters of cases without specifying their size (number of areas) or geographic location in advance. Their statistical significance is tested while adjusting for the multiple testing inherent in such a procedure. However, as is shown in this work, this adjustment is not done in an even manner for all possible cluster sizes. RESULTS: A modification is proposed to the usual inference test of the spatial scan statistic, incorporating additional information about the size of the most likely cluster found. A new interpretation of the results of the spatial scan statistic is done, posing a modified inference question: what is the probability that the null hypothesis is rejected for the original observed cases map with a most likely cluster of size k, taking into account only those most likely clusters of size k found under null hypothesis for comparison? This question is especially important when the p-value computed by the usual inference process is near the alpha significance level, regarding the correctness of the decision based in this inference. CONCLUSIONS: A practical procedure is provided to make more accurate inferences about the most likely cluster found by the spatial scan statistic.

Nonparametric intensity bounds for the delineation of spatial clusters.

BACKGROUND: There is considerable uncertainty in the disease rate estimation for aggregated area maps, especially for small population areas. As a consequence the delineation of local clustering is subject to substantial variation. Consider the most likely disease cluster produced by any given method, like SaTScan, for the detection and inference of spatial clusters in a map divided into areas; if this cluster is found to be statistically significant, what could be said of the external areas adjacent to the cluster? Do we have enough information to exclude them from a health program of prevention? Do all the areas inside the cluster have the same importance from a practitioner perspective? RESULTS: We propose a method to measure the plausibility of each area being part of a possible localized anomaly in the map. In this work we assess the problem of finding error bounds for the delineation of spatial clusters in maps of areas with known populations and observed number of cases. A given map with the vector of real data (the number of observed cases for each area) shall be considered as just one of the possible realizations of the random variable vector with an unknown expected number of cases. The method is tested in numerical simulations and applied for three different real data maps for sharply and diffusely delineated clusters. The intensity bounds found by the method reflect the degree of geographic focus of the detected clusters. CONCLUSIONS: Our technique is able to delineate irregularly shaped and multiple clusters, making use of simple tools like the circular scan. Intensity bounds for the delineation of spatial clusters are obtained and indicate the plausibility of each area belonging to the real cluster. This tool employs simple mathematical concepts and interpreting the intensity function is very intuitive in terms of the importance of each area in delineating the possible anomalies of the map of rates. The Monte Carlo simulation requires an effort similar to the circular scan algorithm, and therefore it is quite fast. We hope that this tool should be useful in public health decision making of which areas should be prioritized.

Hepatitis B, hepatitis C and HIV transfusion-transmitted infections in the 21st century.

In the past, transfusion-transmitted virus (TTV) infections were not uncommon. In recent years with advanced technologies and improved donor screening, the risk of viral transfusion transmission has been markedly reduced. Hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) have all shown marked reduction in transmission rates. However, the newer technologies, including nucleic acid technology (NAT) testing, have affected the residual rates differently for these virally transmitted diseases. Zero risk, which has been the goal, has yet to be achieved. False negatives still persist, and transmissions of these viruses still occur, although rarely. It is known that HBV serological testing misses some infected units; likewise, HBV NAT-negative units have also been known to transmit the virus. Similarly, HIV minipool NAT-negative units have transmitted HIV, as recently as 2007; likely, these transmissions would have been prevented with single-unit NAT testing. Newer technologies, such as pathogen inactivation (PI), will (ideally) eliminate these falsely test negative components, regardless of the original testing method used for detecting the viruses.

Production of biodiesel via enzymatic ethanolysis of the sunflower and soybean oils: modeling.

Biodiesel has become attractive due to its environmental benefits compared with conventional diesel. Although the enzymatic synthesis of biodiesel requires low thermal energy, low conversions of enzymatic transesterification with ethanol (ethanolysis) of oils to produce biodiesel are reported as a result of deactivation of the enzyme depending on the reaction conditions. The synthesis of biodiesel via enzymatic ethanolysis of sunflower and soybean oils was investigated. Kinetic parameters for the overall reactions were fitted to experimental data available in the literature with the Ping Pong Bi-Bi mechanism including the inhibition effect of the ethanol on the activity of lipase Novozyme 435. The model was applied to a batch reactor and the experimental conversions were successfully reproduced. The modeling of a semibatch reactor with continuous addition of ethanol was also performed and the results showed a reduction of roughly 3 h in the reaction time in comparison with the batch-wise operation.

Survey of current practice for prevention of transfusion-transmitted cytomegalovirus in the United States: leucoreduction vs. cytomegalovirus-seronegative.

BACKGROUND AND OBJECTIVES: Cytomegalovirus (CMV) is a significant pathogen transmissible through blood transfusion that can have devastating effects on immunocompromised patients. Current transfusion practice provides two choices for transfusion of cellular blood components that reduce the risk of transfusion-transmitted CMV (TT-CMV): blood components collected from CMV seronegative donors and leucocyte-reduced (LR) blood components. MATERIALS AND METHODS: A web-based survey was designed and administered to AABB physician members in April 2007 to collect information regarding current blood banking and clinical practices for prevention of TT-CMV in the United States. RESULTS: Individuals representing 183 different institutions completed the entire survey (an institutional response rate of 32.5%). Sixty-five percent of respondents indicated that their institution considered that CMV-seronegative and LR products are equally effective in preventing TT-CMV. When analyzed by institutional type, academic institutions and community hospitals were more likely to subscribe to the premise that LR blood components are equally effective at preventing TT-CMV, than were community blood centres and government institutions. However, reported practices for specific patient populations did not match this view of equivalence between CMV-seronegative and LR products with many patient populations preferentially receiving CMV-seronegative components. Fetal and neonatal populations were more likely than other patient populations to receive CMV-seronegative products to reduce the risk of TT-CMV. CONCLUSION: There is wide variability in transfusion practices to reduce the risk of TT-CMV. Lack of a consensus approach may reflect the conflicting data that exist in the literature as well as adherence to longstanding practice.

Assessment of the role of heparan sulfate in high molecular weight kininogen binding to human umbilical vein endothelial cells.

The assembly and activation of the kinin forming system components on human umbilical vein endothelial cells (HUVEC) have been studied in great detail. Proteins such as gC1qR, cytokeratin-1 and u-PAR have been identified to be responsible for Zn2+-dependent binding of high molecular weight kininogen (HK) to HUVEC. Heparan sulfate has also been shown to have a major role in Zn2+-dependent binding of HK to the endothelial cell line, Ea.hy 926. In this study, we have analyzed the possible contribution of heparan sulfate to high molecular weight kininogen binding to HUVEC using multiple approaches. The presence of heparan sulfate on HUVEC was analyzed by staining with an antibody specific for heparan sulfate. Incubation of the cells with bacterial heparinases removed the heparan sulfate from the cell surface to the level seen with a control antibody, however, the Zn2+-dependent binding of HK was not affected. Further, blocking of heparan sulfate with a specific antibody to heparan sulfate even after digestion with heparinases did not reduce HK binding whereas antibodies to the proteins gC1qR and cytokeratin-1 consistently reduced the binding of HK to the endothelial cells. The binding intensities of FITC-labeled HK were similar in heparinase-treated and -untreated HUVEC. The rate of kallikrein formation by the assembly of factor XII, HK and PK were similar in both heparinase-treated and non-treated HUVEC. All of these data indicate that heparan sulfate does not contribute significantly to HK binding to HUVEC.

HIV transmissions from a window-period platelet donation.

Recently, blood centers began investigational testing for HIV RNA by pooled nucleic acid testing (NAT). A 35-year-old frequent platelet donor tested HIV p24 antigen positive, antibody negative before implementation of NAT. He made 2 platelet donations (day -4 and -11) immediately before testing positive for HIV. The donor's HIV seroconversion was monitored, and stored samples were tested retrospectively for HIV RNA. Platelet recipients were tested for HIV infection. The day -4 sample tested positive for HIV RNA by pooled and individual sample NAT. The day -11 sample tested negative for HIV RNA by both NAT tests. The 2 recipients of the day -4 platelets tested HIV RNA and p24 antigen positive. The recipient of the day -11 platelets could not be tested because he had died. HIV NAT would have prevented transmission of HIV had it been available at the time of this donor's HIV seroconversion.

Suppression of Cox-2 and TNF-alpha mRNA in endotoxin tolerance: effect of cycloheximide, antinomycin D, and okadaic acid.

Lipopolysaccharide (LPS)-tolerant human promonocytic THP-1 cells produce decreased levels of inflammatory mediators such as eicosanoids and tumor necrosis factor alpha (TNFalpha) in response to LPS. We hypothesized that transcriptional repression by newly synthesized proteins may be a mechanism for the reduced cellular response to a secondary challenge with LPS. THP-1 cells were desensitized after a 3.5 h or 20 h pre-exposure to LPS (1 microg/mL) and subsequently challenged with LPS (10 microg/mL). In cells rendered tolerant by exposure to LPS for 20 h, LPS-induced expression of cyclooxygenase (Cox)-2 and TNFalpha mRNA was suppressed. Cycloheximide (10 microM) prevented the transcriptional down-regulation of Cox-2 mRNA and to a lesser extent, TNFalpha mRNA, in LPS-tolerant cells. Transcriptional arrest with actinomycin D stabilized steady-state expression of Cox-2 mRNA in naive and tolerant cells but destabilized TNFalpha mRNA expression in LPS-tolerant cells. The observation that in naive cells Cox-2 and TNFalpha mRNA levels subside at 3 to 4 h after LPS (10 microg/mL or 1 microg/mL) suggested that LPS tolerance may occur earlier. Therefore, in subsequent experiments, the effect of LPS pretreatment for only 3.5 h was examined. This abbreviated tolerance regimen diminished secondary LPS-induced Cox-2 mRNA expression but had a lesser effect on TNFalpha mRNA expression. However, cycloheximide augmented both Cox-2 and TNFalpha mRNA expression in this group. Also, the serine/threonine phosphatase inhibitor okadaic acid augmented Cox-2 and TNFalpha mRNA expression in the LPS-tolerant cells. Although LPS-induced TNFalpha production in LPS-tolerant cells was suppressed relative to the naive cells, okadaic acid induced comparable levels of TNFalpha in tolerant and naive cells. These findings support the concept that LPS tolerance is associated with induction of proteins that alter expression of certain genes. Expression of Cox-2 mRNA appears to be particularly sensitive to down-regulation and, to a lesser extent, TNFalpha mRNA. However, this seems to vary depending on the LPS pretreatment regimen. The ability of a phosphatase inhibitor to induce TNFalpha and expression of Cox-2 and TNFalpha mRNA in LPS tolerance suggests that there may be alterations in phosphorylation status of signaling pathways, transcriptional mechanisms, or post-transcriptional mRNA stability.