Efetividade do tratamento endodôntico em sessão única x múltiplas sessões na periodontite apical pós-tratamento: sinais e sintomas; carga microbiana; níveis de endotoxinas e ácido lipoteicóico e volumetria da destruição óssea periapical por TCFC / Effectiveness of one-visit and two-visits endodontic treatment in posttreatment apical periodontitis: signs and symptoms; microbial load; endotoxin and lipoteichoic acid levels and bone destruction analysis by CBCT in 18-months follow-up

Os objetivos deste estudo foram: a) avaliar o sucesso clínico e por tomografia computadorizada de feixe cônico (TCFC) após 18 meses do retratamento em sessão única (RU) e múltiplas sessões (RM); b) monitorar carga (UFC/mL) e perfil microbiano; níveis de endotoxinas (EU/mL) e de ácido lipoteicóico [LTA (pg/mL)] em dentes com periodontite apical pós-tratamento (PAPT) submetidos a RU e RM; c) comparar a sintomatologia pós-operatória pela escala visual analógica de dor (EVA) em RU e RM; d) comparar os tratamentos quanto à redução de UFC/mL, EU/mL, LTA (pg/mL) e a volumetria da lesão periapical (LP) e) avaliar as correlações e associações entre os dados obtidos no estudo. Seguindo critérios de inclusão e não-inclusão, foram selecionados 40 dentes com PAPT, divididos em dois grupos: RM e RU. Os pacientes foram submetidos à TCFC. Foram realizadas coletas do conteúdo do canal radicular: após remoção do material obturador (S1), após preparo biomecânico (PBM) (S2) e após MIC (S3). Ao final da sessão, foi fornecida a EVA para avaliação da sintomatologia dolorosa apresentada nos períodos de 24hs, 48hs e 7 dias. O conteúdo dos canais radiculares foi avaliado por cultura microbiológica, perfil microbiano por Checkerboard DNA-DNA hybridization, níveis de endotoxinas pelo teste Lisado Amebócito de Limulus e de ácido lipoteicóico pelo ensaio de ELISA. Foi realizada a volumetria da destruição óssea periapical inicial e final por TCFC pelo software ITK snap. Os dados foram analisados estatisticamente. Os níveis de UFC/mL, EU/mL e LTA (pg/mL) diminuíram após o PBM, havendo a manutenção dos mesmos após o uso de MIC (p>0,05). As coletas apresentaram valores iniciais (S1) maiores que os residuais (antes da obturação) para os parâmetros avaliados. Quanto aos valores residuais, foram encontrados maiores valores de UFC/mL em RM e menores valores de LTA no mesmo grupo (p<0,05); porém sem diferença entre os grupos RU e RM quanto aos valores residuais de EU/mL. Os micro-organismos Gram-negativos mais encontrados foram o F. nucleatum, C. rectus, C. gingivalis, P. gingivalis, P. intermedia e L. buccalis, sendo os Gram-positivos: E. faecalis, E. faecium, S. constellatus e S. mitis. Não houve diferença entre os grupos RU e RM quanto à sintomatologia pósoperatória (p>0,05) pela EVA. O volume inicial das LP variou de 10 a 470 mm³, sendo que o final variou de 0 a 48 mm³. Houve redução da média dos volumes de 84,71% em RU e de 90.56% em RM, sendo esta redução significante em ambos os grupos (p< 0,0001), mas sem diferença estatística entre eles (p=0,9117). No período de 18 meses após o retratamento endodôntico não houve diferenças na regressão das lesões e análise dos demais sinais e sintomas entre RU e RM(AU)

This study aimed to: a) evaluate the success rates of one-visit (1-visit) and two-visit (2- visit) endodontic retreatment after 18 months by cone-beam computed tomography (CBCT) and analyze clinical signs and symptoms; b) monitor and compare the microbial load and profile; levels of endotoxins and lipoteichoic acid in teeth with posttreatment apical periodontitis (PTAP); c) compare the postoperative pain between the groups by the visual analog pain scale (VAS); d) correlate the variables with themselves. Forty teeth with PTAP were selected and submitted to CBCT. The sampling procedure was performed: after filling material removal (S1), after biomechanical preparation (S2), and after the intracanal medication placement (S3). VAS was provided for assessment of the postoperative pain at different time-points (24 h, 48 h, and 7 days). Root canal contents were submitted to microbiological assessment by culture technique (CFU/mL) and Checkerboard DNA-DNA hybridization, and the determination of LPS (UE/mL) and LTA (pg/mL) levels by Limulus amebocyte lysate and enzyme-linked immunosorbent assays, respectively. Periapical lesions volumes were obtained by the ITK snap program. Data were statistically analyzed. CFU/mL, UE/mL, and LTA (pg/mL) decreased after biomechanical preparation, with their maintenance after the use of intracanal medication (p>0.05). All root canal samples had their baseline values (S1) higher than those found before the root canal obturation (residual values) for all parameters. Regarding the residual values, higher values of CFU/mL and lower values of LTA were found in 2-visit groups (p<0.05). The most frequently Gram-negative microorganisms found during sampling procedures were F. nucleatum, C. rectus, C. gingivalis, P. gingivalis, P. intermedia and L. buccalis, the Gram-positive ones were E. faecalis, E. faecium, S constellatus and S. mitis. There was no difference between the 1 or 2-visit treatments regarding postoperative symptoms (p>0.05). The initial volume of the periapical lesions ranged from 10 to 470 mm³, and the final ones ranged from 0 to 48 mm³. A significant reduction of the periapical lesion was observed after 18-months in both groups (p<0.0001), but no statistical difference was found between them (p>0.05). No significant differences were observed in the outcome of the two modalities of endodontic retreatment(AU)

Indoor endotoxin, proximity to a major roadway, and severe asthma exacerbations among children in Puerto Rico.

BACKGROUND: Few studies have examined concurrent exposure to household endotoxin and traffic-related air pollution in relation to childhood asthma, yet both factors are associated with asthma outcomes. OBJECTIVE: To examine whether proximity to a major roadway (a traffic-related air pollution proxy) modifies the estimated effects of indoor endotoxin on asthma outcomes in children. METHODS: Cross-sectional study of 200 children with asthma (ages, 6-14 years) living in Puerto Rico. Residential distance to a major roadway was calculated as the distance from the participant's residential US census block centroid to the nearest major road. The outcomes of interest were severe asthma exacerbations, missed school days for asthma, atopy, lung function, and bronchodilator response (BDR). Logistic, linear, or negative binomial regression was used for the multivariable analysis. RESULTS: In the multivariable analysis, there was an interaction between indoor endotoxin and residential distance to a roadway on severe asthma exacerbations (P = .02) and BDR (P = .07). In an analysis stratified by distance to a roadway, each log10-unit increase in endotoxin was associated with 4.21 times increased odds of severe asthma exacerbations among children living within 499 m (the lower 3 quartiles of residential distance) to a road (95% confidence interval, 1.5-12.0). Among subjects living further than 499 m away from a roadway, each log10-unit increase in endotoxin was associated with reduced odds of severe asthma exacerbations (odds ratio, 0.03; 95% confidence interval, 0.001-0.67). Similar but less substantial findings were observed for BDR. CONCLUSION: Our findings suggest that residential proximity to a major road modifies the estimated effect of endotoxin on severe asthma exacerbations in children.

Reacciones adversas relacionadas con endotoxinas bacterianas recibidas por el Sistema Nacional de Farmacovigilancia (2006-2017) / Adverse reactions related to bacterial endotoxins received by the National Pharmacovigilance System

En el presente estudio se relevaron los efectos adversos producidos por medicamentos (metotrexato, metronidazol y gluconato de calcio) contaminados con endotoxinas bacterianas y registrados en la base de desvíos de la calidad del Sistema Nacional de Farmacovigilancia durante el período 2006-2017. De acuerdo a las notificaciones recibidas y analizadas, las reacciones adversas más frecuentes observadas fueron fiebre, temblores, escalofríos, sudoración, disnea súbita, vómitos y taquicardia, entre otras. La aparición de estas reacciones fue cercana a las 3 horas luego de la administración, por la vía intratecal, mientras que, por la vía intravenosa, los tiempos variaron entre los 10 y 60 minutos de iniciada la administración de los medicamentos, con una duración entre 5-20 minutos hasta 2 horas. Estos signos y síntomas, producidos por el uso de productos contaminados, difieren de aquellos eventos adversos causados por la acción propia de los fármacos involucrados, por lo cual destacamos el rol fundamental de los profesionales en el reconocimiento de problemas o fallas en la calidad del producto.

The adverse effects produced by medications (methotrexate, metronidazole and calcium gluconate) contaminated with bacterial endotoxins registered in the National Pharmacovigilance System database of quality deviations were surveyed in the period 2006-2017. According to the notifications that were received and analyzed, the most frequent adverse reactions observed were fever, trembling, chills, sweating, sudden dyspnea, vomiting and tachycardia, among others. The appearance of the reactions was close to 3 hours for the intrathecal route, whereas for the intravenous route, the time was between 10 and 60 minutes after the administration, lasting between 5-20 minutes up to 2 hours. These signs and symptoms that derived from the use of contaminated products differ from those adverse events caused by the typical action of the molecule of a drug, therefore, we emphasize the fundamental role of the professionals for the awareness of problems or failures in the quality of the product.

Yield Losses in Transgenic Cry1Ab and Non-Bt Corn as Assessed Using a Crop-Life-Table Approach.

In this study, we constructed crop life tables for Bacillus thuringiensis Berliner (Bt) Cry1Ab and non-Bt corn hybrids, in which yield-loss factors and abundance of predaceous arthropods were recorded during 2 yr at two locations. Corn kernel/grain was the yield component that had the heaviest losses and that determined the overall yield loss in the corn hybrids across years and locations. Yield losses in both corn hybrids were primarily caused by kernel-destroying insects. Helicoverpa zea (Boddie) and Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) were the key loss factors at one location, while at the other, the key loss factor was the silk fly larvae, Euxesta spp. (Diptera: Ulidiidae). Although the realized yield of corn grains was not different (P > 0.05) between Cry1Ab and non-Bt corn hybrids, the Bt corn hybrid reduced (P < 0.05) the damage by H. zea and S. frugiperda in three of the four field trials, particularly at the location where Lepidoptera were the key loss factors. As expected, no reduction in the abundance of predaceous arthropods was observed in Cry1Ab corn fields. Various species of natural enemies were recorded, particularly the earwig Doru luteipes (Scudder) (Dermaptera: Forficulidae), which was the most abundant and frequent predaceous insect. These results indicate that integration of pest management practices should be pursued to effectively minimize losses by kernel-destroying insects during corn reproductive stages when growing non-Bt or certain low-dose Bt corn cultivars for fall armyworm and corn earworm, such as those producing Cry1Ab or other Cry toxins.

Infecções endodônticas primária x secundária: perfil microbiano, níveis de endotoxinas e ácido lipoteicóico, sinais e sintomas / Primary x secondary endodontic infections: microbial profile, endotoxins and lipoteichoic acid levels, signs and symptoms

Esta pesquisa clínica teve como objetivo comparar a carga e composição microbiana bem como as concentrações de LPS e LTA encontradas na infecção endodôntica primária (IEP) e na infecção endodôntica secundária (IES). Além disso, a correlação desses achados com características clínicas e tomográficas também foram investigadas. Sessenta dentes de pacientes com IEP (31) e IES (29) foram submetidos à avaliação clínica e tomográfica, seguido do tratamento endodôntico ou retratamento. Amostras foram coletadas de cada canal radicular utilizando cones de papel. Logo após a abertura coronária (IEP) ou após a desobturação dos canais (IES) o conteúdo coletado foi submetido à técnica de cultura microbiológica para determinar a carga microbiana de bactérias anaeróbias e ao método Checkerboard DNA-DNA hybridization para investigação de espécies bacterianas presentes. O teste de Lisado de Amebócito de Limulus e o Ensaio de Imunoabsorção Enzimática foram utilizados para quantificar os níveis de LPS e LTA. Os dados obtidos foram correlacionados com os achados clínicos e tomográficos. Maiores quantidades de bactérias cultiváveis e de LPS foram encontradas na IEP (p < 0,05). Não houve diferença nos níveis de LTA entre IEP e IES (p > 0,05). A mediana de espécies por canal radicular encontrada na IEP foi de 9 espécies e na IES foi de 22 (p < 0,05). As espécies bacterianas mais prevalentes detectadas na IEP foram P. gingivalis (14/31) e S. intermedius (14/31). Na IES, as espécies mais prevalentes foram P. gingivalis (21/29) e C. rectus (20/29). LPS foi correlacionado positivamente com um maior volume da lesão periapical (p < 0,05). Níveis de LTA não foram relacionados a sinais e sintomas ou ao volume da lesão periapical (p > 0,05). Concluiu-se que dentes com IEP tiveram maiores quantidades de carga microbiana e de LPS do que os dentes com IES. Uma maior quantidade de LPS foi correlacionada positivamente com um maior volume de destruição óssea periapical. Uma ampla interação de espécies bacterianas específicas resultou em diferentes características clínicas(AU)

This clinical research aimed to compare the microbial load and composition as well as the LPS and LTA concentrations found in Primary Endodontic Infection (PEI) and Secondary Endodontic Infection (SEI). In addition, the correlation of these findings with clinical and tomographic features was also investigated. Sixty patients' teeth with PEI (31) and SEI (29) were submitted to clinical and tomographic assessment, followed by endodontic treatment or retreatment. Samples were taken from each root canal using paper points. After the coronary opening (PEI) or after the removal of root filling material (SEI), the collected samples were submitted to the microbiological culture technique to determine the microbial load of anaerobic bacteria and to the Checkerboard DNA-DNA hybridization method for investigation of present bacterial species. The Limulus amebocyte lysate assay and enzyme-linked immunosorbent assay were used to quantify LPS and LTA levels. The data obtained were correlated with clinical and tomographic findings. A higher number of cultivable bacteria and LPS was found in PEI (p < 0.05). There was no difference in LTA levels between PEI and SEI (p> 0.05).The median number of species per root canal found in PEI was 9 and 22 in SEI (p < 0.05). The most prevalent bacterial species detected in PEI were P. gingivalis (14/31) and S. intermedius (14/31). In SEI, the most prevalent species were P. gingivalis (21/29) and C. rectus (20/29). LPS was positively correlated with a larger periapical lesion volume (P < 0.05). LTA levels were not related to signs and symptoms or periapical lesion volume (p> 0.05). It was concluded teeth with PEI had higher contents of microbial load and LPS than teeth with SEI. However, a more diverse microbiota was found in SEI than that of PEI. Higher content of LPS was positively correlated with larger periapical bone destruction. A widely interaction of specific microbial species resulted in different clinical features(AU)

Avaliação clínica do efeito de diferentes medicações intracanais nos níveis de resolvinas e lipoxinas periapicais: novas perspectivas endodônticas / Clinical evaluation of the effect of different intracanal medications on levels of periapical resolvins and lipoxins: new endodontic perspectives

RESUMO O objetivo do estudo foi avaliar o efeito da terapia endodôntica utilizando NaOCl 2,5% e medicações intracanais a base de hidróxido de cálcio e de da Nacetil cisteína (NAC) na desinfecção dos canais radiculares, na redução de endotoxinas e na estimulação de liberação dos mediadores lipídicos Resolvina E1, D2 (RvE1, RvD2) e Lipxina A4 (LxA4). Foram selecionados quarenta dentes unirradiculares, com infecção endodôntica primária e periodontite apical. Os dentes foram aleatoriamente divididos em 3 grupos de acordo com a medicação intracanal a ser utilizada: G1: soro fisiológico + Ca(OH)2 (n=14),G2: N-Acetil Cisteína (n=13), G3: clorexidina (CLX) + Ca(OH)2 (n=14). Amostras bacterianas e de endotoxinas foram coletadas do canal radicular, após abertura coronária, após preparo dos canais com limas reciprocantes (Reciproc) selecionadas de acordo com o diâmetro do canal radicular e a solução irrigante (NaOCl 2,5%) e após medicação intracanal. A análise da atividade antimicrobiana foi através da contagem de unidades formadoras de colônias (UFC/mL) de microrganismos remanescentes no canal radicular e quantificação de endotoxinas (EU/mL) através do teste Limulus Amebocyte Lysate ­ LAL. Após o preparo biomecânico (PBM) os dentes foram preenchidos com as medicações intracanais de acordo com os grupos por quatorze dias. O fluído intersticial foi coletado após preparo dos canais e após 14 dias de medicação e a quantificação dos mediadores lipídicos (RvE1, RvD2, LxA4) foi realizada através do teste imunoenzimático (ELISA). Os valores obtidos foram tabulados, analisados pelo software GraphPad Prism 6.01 e submetidos aos testes de normalidade de Kolmogorov-Smirnov e Lilliefors e em seguida aos testes Kruskal-Wallis e Dunn e Anova one-way e Tukey (P <0.05). Para carga microbiana, o PBM reduziu em média 96% dos microrganismos aeróbios e 92,5% dos anaeróbios em todos os grupos (p<0,05). A utilização de medicação intracanal à base de Ca(OH)2 manteve os níveis atingidos com o PBM (p>0,05), porém no grupo NAC houve aumento de bactérias. Os níveis de endotoxinas diminuíram após o preparo dos canais e o uso das medicações reduziu o nível de endotoxinas porém sem diferença estatística em relação à redução alcançada com o preparo biomecânico. Quanto aos mediadores lipídicos, somente no grupo NAC foi observado aumento de Rv D2. Concluiu-se que o PBM reduziu significantemente o número de bactérias e a quantidade de endotoxinas do canal, e as medicações à base de Ca(OH)2 foram eficientes na eliminação de microrganismos do canal, entretanto NAC não foi eficaz para a redução da carga microbiana; As três medicações avaliadas foram capazes de atuar no LPS bacteriano. Somente a NAC foi capaz de influenciar positivamente esse resultado com o aumento de RvD2 pós 14 dias da medicação. Os mediadores lipídicos têm grande potencial para uso nos diversos tipos de tratamento endodôntico(AU)

ABSTRACT The aim of this study was to evaluate the effect of endodontic therapy using 2.5% NaOCl and intracane medications based on calcium hydroxide and Nacetyl cysteine (NAC) on root canal disinfection, reduction of endotoxins and stimulation of release of the lipid mediators Resolvin E1, D2 (RvE1, RvD2) and Lipoxin A4 (LxA4). Forty uniradicular teeth were selected, with primary endodontic infection and apical periodontitis. The teeth were randomly divided into 3 groups according to the intracanal medication to be used: G1: saline + Ca (OH) 2 (n = 14), G2: N-Acetyl Cysteine (n = 13), G3: chlorhexidine CLX) + Ca (OH) 2 (n = 14). Bacterial and endotoxin samples were collected from the root canal after coronary opening after preparation of the channels with reciprocal files (Reciproc) selected according to the root canal diameter and the irrigating solution (NaOCl 2.5%) and after intracanal medication. The analysis of the antimicrobial activity was by count of colony forming units (CFU / mL) of microorganisms remaining in the root canal and quantification of endotoxins (EU / mL) using the Limulus Amebocyte Lysate - LAL test. After biomechanical preparation (PBM) the teeth were filled with intracanal medications according to the groups for fourteen days. The interstitial fluid was collected after preparation of the channels and after 14 days of medication and quantification of the lipid mediators (RvE1, RvD2, LxA4) was performed through the enzymelinked immunosorbent assay (ELISA). The values obtained were tabulated, analyzed by GraphPad Prism 6.01 software and submitted to Kolmogorov Smirnov and Lilliefors normality tests, followed by Kruskal-Wallis and Dunn and Anova one-way and Tukey tests (P <0.05). For microbial load, PBM reduced on average 96% of aerobic microorganisms and 92.5% of anaerobes in all groups (p <0.05). The use of intracanal medication based on Ca (OH) 2 maintained levels reached with PBM (p> 0.05), but in the NAC group there was an increase of bacteria. The endotoxin levels decreased after the preparation of the canals and the use of the medications reduced the endotoxin level, but without statistical difference in relation to the reduction achieved with the biomechanical preparation. As for the lipid mediators, only in the NAC group an increase of D2 Rv was observed. It was concluded that PBM significantly reduced the number of bacteria and the amount of endotoxins in the canal, and Ca (OH) 2-based medications were efficient in the elimination of microorganisms from the canal, however NAC was not effective for reducing the load microbial; The three medications evaluated were able to act on bacterial LPS. Only the NAC was able to positively influence this result with the increase of RvD2 after 14 days of the medication. Lipid mediators have great potential for use in various types of endodontic treatment(AU)

Exposure to high endotoxin concentration increases wheezing prevalence among laboratory animal workers: a cross-sectional study.

BACKGROUND: Endotoxin from Gram-negative bacteria are found in different concentrations in dust and on the ground of laboratories dealing with small animals and animal houses. METHODS: Cross-sectional study performed in workplaces of two universities. Dust samples were collected from laboratories and animal facilities housing rats, mice, guinea pigs, rabbits or hamsters and analyzed by the "Limulus amebocyte lysate" (LAL) method. We also sampled workplaces without animals. The concentrations of endotoxin detected in the workplaces were tested for association with wheezing in the last 12 months, asthma defined by self-reported diagnosis and asthma confirmed by bronchial hyperresponsiveness (BHR) to mannitol. RESULTS: Dust samples were obtained at 145 workplaces, 92 with exposure to animals and 53 with no exposure. Exposed group comprised 412 subjects and non-exposed group comprised 339 subjects. Animal-exposed workplaces had higher concentrations of endotoxin, median of 34.2 endotoxin units (EU) per mg of dust (interquartile range, 12.6-65.4), as compared to the non-exposed group, median of 10.2 EU/mg of dust (interquartile range, 2.6-22.2) (p < 0.001). The high concentration of endotoxin (above whole sample median, 20.4 EU/mg) was associated with increased wheezing prevalence (p < 0.001), i.e., 61 % of workers exposed to high endotoxin concentration reported wheezing in the last 12 months compared to 29 % of workers exposed to low endotoxin concentration. The concentration of endotoxin was not associated with asthma report or with BHR confirmed asthma. CONCLUSION: Exposure to endotoxin is associated with a higher prevalence of wheezing, but not with asthma as defined by the mannitol bronchial challenge test or by self-reported asthma. Preventive measures are necessary for these workers.

Study of the expression of GABA(A) receptor in rats during acute lung injury caused by endotoxin.

The objective of the present study was to investigate the role of γ-aminobutyric acid type A receptor (GABA(A)R) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. Thirty-two male wistar rats were randomly divided into four groups. Rats in the GABA group were pretreated with LPS and GABA, while those in the bicuculline (BIC) group were pretreated with LPS and bicuculline. We assessed the arterial blood gas, dry/wet ratio, and the level of tumor necrosis factor-α (TNF-α), IL-6, malondialdehyde, and superoxide dismutase 6 h after the immunization. Paraffin sections of samples were detected using the steptavidin-peroxidase method. Protein expression was detected using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blotting. PaO2 in the LPS group was significantly lower than that in the control rats. Activation of GABA-mediated signaling by GABA increased the expression of GABA(A)R in airway bronchial and alveolar epithelial cells. Blockade of the GABA(A)R by bicuculline limited the expression of this receptor. The GABA group rats had higher levels of tissue TNF-α and IL-6 than in ALI rats and control rats. The BIC group rats demonstrated an opposite expression level compared to the GABA group rats. Our results suggest that the GABA(A)R could aggravate the inflammatory response syndrome and oxidative stress in the lungs and play an essential role in LPS-induced acute lung injury. It provides a novel method to study the incidence and mortality of ALI during the peroperative period.

Food safety assessment of Cry8Ka5 mutant protein using Cry1Ac as a control Bt protein.

Cry8Ka5 is a mutant protein from Bacillus thuringiensis (Bt) that has been proposed for developing transgenic plants due to promising activity against coleopterans, like Anthonomus grandis (the major pest of Brazilian cotton culture). Thus, an early food safety assessment of Cry8Ka5 protein could provide valuable information to support its use as a harmless biotechnological tool. This study aimed to evaluate the food safety of Cry8Ka5 protein following the two-tiered approach, based on weights of evidence, proposed by ILSI. Cry1Ac protein was used as a control Bt protein. The history of safe use revealed no convincing hazard reports for Bt pesticides and three-domain Cry proteins. The bioinformatics analysis with the primary amino acids sequence of Cry8Ka5 showed no similarity to any known toxic, antinutritional or allergenic proteins. The mode of action of Cry proteins is well understood and their fine specificity is restricted to insects. Cry8Ka5 and Cry1Ac proteins were rapidly degraded in simulated gastric fluid, but were resistant to simulated intestinal fluid and heat treatment. The LD50 for Cry8Ka5 and Cry1Ac was >5000 mg/kg body weight when administered by gavage in mice. Thus, no expected relevant risks are associated with the consumption of Cry8Ka5 protein.

Anti-inflammatory mechanisms of the annexin A1 protein and its mimetic peptide Ac2-26 in models of ocular inflammation in vivo and in vitro.

Annexin A1 (AnxA1) is a protein that displays potent anti-inflammatory properties, but its expression in eye tissue and its role in ocular inflammatory diseases have not been well studied. We investigated the mechanism of action and potential uses of AnxA1 and its mimetic peptide (Ac2-26) in the endotoxin-induced uveitis (EIU) rodent model and in human ARPE-19 cells activated by LPS. In rats, analysis of untreated EIU after 24 and 48 h or EIU treated with topical applications or with a single s.c. injection of Ac2-26 revealed the anti-inflammatory actions of Ac2-26 on leukocyte infiltration and on the release of inflammatory mediators; the systemic administration of Boc2, a formylated peptide receptor (fpr) antagonist, abrogated the peptide's protective effects. Moreover, AnxA1(-/-) mice exhibited exacerbated EIU compared with wild-type animals. Immunohistochemical studies of ocular tissue showed a specific AnxA1 posttranslational modification in EIU and indicated that the fpr2 receptor mediated the anti-inflammatory actions of AnxA1. In vitro studies confirmed the roles of AnxA1 and fpr2 and the protective effects of Ac2-26 on the release of chemical mediators in ARPE-19 cells. Molecular analysis of NF-κB translocation and IL-6, IL-8, and cyclooxygenase-2 gene expression indicated that the protective effects of AnxA1 occur independently of the NF-κB signaling pathway and possibly in a posttranscriptional manner. Together, our data highlight the role of AnxA1 in ocular inflammation, especially uveitis, and suggest the use of AnxA1 or its mimetic peptide Ac2-26 as a therapeutic approach.

Respiratory, allergy and eye problems in bagasse-exposed sugar cane workers in Costa Rica.

AIMS: To evaluate bagasse (sugar cane fibres) and microbiological exposure among sugar cane refinery workers in Costa Rica and its relationships with respiratory, allergy and eye problems. METHODS: Ventilatory lung function and total serum IgE were measured in 104 sugar cane workers in five departments at one refinery before the harvesting season, and repeated for 77 of the workers at the end of the season. Information on the prevalence of respiratory and other symptoms was collected with a standardised questionnaire. During the harvesting season, inhalable dust, endotoxin and mould levels were measured among 74 randomly selected sugar cane workers across departments. RESULTS: During the harvesting season, dust levels were relatively high in some departments, while endotoxin and mould levels were around background levels. Workers' ventilatory lung function differed between departments before, but not during the harvesting season or between seasons. During the harvesting season, the prevalence of wheeze and eye problems almost doubled in workers exposed to bagasse and other types of dust, whereas shortness of breath and rhinitis increased only in bagasse-exposed workers. Reporting wheeze and shortness of breath was positively associated with the number of years working at the refinery, suggesting a long-term health effect. CONCLUSION: In this refinery, the differences in workers' ventilatory lung function before the harvesting season are unlikely to be explained by bagasse exposure. However, the increase in reported symptoms (wheeze, shortness of breath, eye problems and rhinitis) over the season is likely due to irritation by dust, in particular bagasse, rather than microbiological agents.

Effects of Japanese herbal medicine Inchin-ko-to on endotoxin-induced cholestasis in the rat.

BACKGROUND/OBJECTIVE: Inchin-ko-to (ICKT) is an herbal medicine used in Japan to treat jaundice and liver fibrosis.We investigated the effect of oral ICKT supplementation on endotoxin-induced cholestasis in the rat. MATERIAL AND METHODS: Lipopolysaccharide (LPS) injection (1 mg/kg body weight i.p.) was used as a model of sepsis-induced cholestasis. Bile flow, biliary bile salt secretion, biliary glutathione secretion and protein expression of the main hepatobiliary transporters Na(+)-taurocholate-cotransporting peptide (Ntcp), multidrug resistance protein 2 (Mrp2) and bile salt export pump (Bsep) were analyzed by conventional techniques in ICKT treated and non-treated animals. RESULTS: Injection of LPS induced a significant decrease of bile flow (-24%), biliary bile salts (-40%) and glutathione excretion (-70%) as well as a significant decrease in Ntcp (-90%) and Mrp2 (-80%) protein levels. ICKT supplementation partially prevented the effects of LPS determining a less intense reduction in bile flow (-10%), a normalization of glutathione excretion as well as a significant increase in Mrp2 protein levels to 60% of the levels observed in control animals. ICKT administration did not modify the effects of LPS on BS secretion or Ntcp protein levels. CONCLUSION: Our data show that oral supplementation of ICKT partially prevents LPS-induced cholestasis by increasing Mrp2 protein levels and biliary glutathione excretion thus increasing bile salt-independent flow.

Histological analysis of the association between formocresol and endotoxin in the subcutaneous tissue of mice.

This study performed a histological analysis of the effect of formocresol associated to endotoxin (LPS) in the subcutaneous connective tissue of mice. Ninety mice were randomly assigned to 3 groups (n=30). Each animal received one plastic tube implant containing endotoxin solution (10 mg/mL), formocresol (original formula) or a mixture of endotoxin and formocresol. The endotoxin and formocresol groups served as controls. The periods of analysis were 7, 15 and 30 days. At each experimental period, tissue samples were collected and submitted to routine processing for histological analysis. Endotoxin and formocresol produced necrosis and chronic inflammation at 7 and 15 days. At 30 days, the endotoxin group showed no necrosis, while in the formocresol group necrosis persisted. The formocresol-endotoxin association produced necrosis and chronic inflammation in the same way as observed with formocresol at all experimental periods. In conclusion, formocresol seems not to be able to inactive the toxic effects of endotoxin in connective tissues.

Histological analysis of the association between formocresol and endotoxin in the subcutaneous tissue of mice

This study performed a histological analysis of the effect of formocresol associated to endotoxin (LPS) in the subcutaneous connective tissue of mice. Ninety mice were randomly assigned to 3 groups (n=30). Each animal received one plastic tube implant containing endotoxin solution (10 mg/mL), formocresol (original formula) or a mixture of endotoxin and formocresol. The endotoxin and formocresol groups served as controls. The periods of analysis were 7, 15 and 30 days. At each experimental period, tissue samples were collected and submitted to routine processing for histological analysis. Endotoxin and formocresol produced necrosis and chronic inflammation at 7 and 15 days. At 30 days, the endotoxin group showed no necrosis, while in the formocresol group necrosis persisted. The formocresol-endotoxin association produced necrosis and chronic inflammation in the same way as observed with formocresol at all experimental periods. In conclusion, formocresol seems not to be able to inactive the toxic effects of endotoxin in connective tissues.

O objetivo deste estudo foi avaliar histologicamente o efeito da associação do formocresol com endotoxina (LPS) em tecido conjuntivo de camundongos. Noventa camundongos foram divididos em três grupos de 30 camundongos cada. Cada camundongo recebeu um implante subcutâneo de tubo plástico contendo solução de endotoxina (10 mg/ml), formocresol (fórmula original), ou uma mistura de formocresol com endotoxina. Os grupos da endotoxina e formocresol foram considerados grupos controle. Os períodos de análise foram 7, 15 e 30 dias. Após os períodos experimentais, os tecidos foram removidos e submetidos a processamento histológico. Os resultados obtidos indicam que a endotoxina e o formocresol produzem necrose e inflamação tecidual crônica aos 7 e 15 dias e aos 30 dias o grupo da endotoxina não mostrava necrose e no grupo do formocresol a necrose persistiu. A combinação formocresol e endotoxina mostrou necrose e inflamação crônica com resultados semelhantes ao do grupo formocresol para todos os períodos experimentais. Pode-se concluir que o formocresol parece não ser capaz de inativar os efeitos tóxicos da endotoxina.

Central heme oxygenase-carbon monoxide pathway participates in the lipopolysaccharide-induced tolerance in rats.

Recently, heme oxygenase-carbon monoxide (HO-CO) pathway has been reported to be involved in the development of lipopolysaccharide (LPS) fever. However, no information exists about its participation in LPS tolerance, which is defined by an attenuation of the febrile response to repeated administrations of LPS. Thus, we tested the hypothesis that HO-CO pathway plays a role in endotoxin tolerance, which was induced by means of three consecutive LPS intraperitoneal injections (i.p.) at 24-h intervals. Body temperature (Tb) was measured by biotelemetry. Induction of the HO pathway using intracerebroventricular (i.c.v.) heme lysinate reversed tolerance, and this effect could be prevented by pretreatment with ODQ [a soluble guanylate cyclase (sGC) inhibitor; i.c.v.]. These results indicate that HO-CO pathway seems to be down-regulated during LPS tolerance, and that CO is the HO product that can prevent LPS tolerance, acting via cGMP. In further support, either biliverdine or iron (the others HO products; i.c.v.) had no effect in LPS-induced tolerance.

[Pathogenesis of portal hypertension]. / Patogénesis de la hipertensión portal.

It is now well established that portal hypertension is not a purely mechanical phenomenon. Primary hemodynamic alterations develop in the hepatic and systemic circulatory systems; these alterations in combination with mechanical factors contribute to the development of portal hypertension. In the hepatic circulation, these hemodynamic alterations are characterized by vasoconstriction and impaired hepatic vasodilatory responses, whereas in the systemic circulation, particularly in the splanchnic bed, vessels are hyperemic with increased flow. Thus, an increase in intrahepatic resistance in conjunction with increased portal venous inflow, mediated through splanchnic dilation, contributes to the development of portal hypertension. The ensuing development of elevated flow and transmural pressure through collateral vessels from the hypertensive portal vasculature into the lower pressure systemic venous circulation accounts for many of the complications, such as bleeding esophageal varices, observed with portal hypertension. The importance of the primary vascular origin of portal hypertension is emphasized by the utility of current therapies aimed at reversing these hemodynamic alterations, such as nitrates, which reduce portal pressure through direct intrahepatic vasodilatation, and ,B blockers and octreotide, which reduce splanchnic vasodilatation and portal venous inflow. New evidence concerning relevant molecular mechanisms of contractile signaling pathways in hepatic stellate cells and the complex regulatory pathways of vasoactive molecules in liver endothelial cells makes a better understanding of these processes essential for developing further experimental therapies for portal hypertension. This article examines the current concepts relating to cellular mechanism that underlie the hemodynamic alterations that characterize and account for the development of portal hypertension.

Patogénesis de la hipertensión portal / Pathogenesis of portal hypertension

It is now well established that portal hypertension is not a purely mechanical phenomenon. Primary hemodynamic alterations develop in the hepatic and systemic circulatory systems; these alterations in combination with mechanical factors contribute to the development of portal hypertension. In the hepatic circulation, these hemodynamic alterations are characterized by vasoconstriction and impaired hepatic vasodilatory responses, whereas in the systemic circulation, particularly in the splanchnic bed, vessels are hyperemic with increased flow. Thus, an increase in intrahepatic resistance in conjunction with increased portal venous inflow, mediated through splanchnic dilation, contributes to the development of portal hypertension. The ensuing development of elevated flow and transmural pressure through collateral vessels from the hypertensive portal vasculature into the lower pressure systemic venous circulation accounts for many of the complications, such as bleeding esophageal varices, observed with portal hypertension. The importance of the primary vascular origin of portal hypertension is emphasized by the utility of current therapies aimed at reversing these hemodynamic alterations, such as nitrates, which reduce portal pressure through direct intrahepatic vasodilatation, and fi blockers and octreotide, which reduce splanchnic vasodilatation and portal venous inflow. New evidence concerning relevant molecular mechanisms of contractile signaling pathways in hepatic stellate cells and the complex regulatory pathways of vasoactive molecules in liver endothelial cells makes a better understanding of these processes essential for developing further experimental therapies for portal hypertension. This article examines the current concepts relating to cellular mechanism that underlie the hemodynamic alterations that characterize and account for the development of portal hypertension.

Actualmente está bien establecido que la hipertensión portal no es un fenómeno puramente mecánico. En esta entidad se presentan alteraciones hemodinámicas primarias en los sistemas circulatorios hepático y sistémico; estas alteraciones en combinación con factores mecánicos, contribuyen al desarrollo de la hipertensión portal. En la circulación hepática, las alteraciones hemodinámicas se caracterizan por vasoconstricción y una respuesta anómala a la vasodilatación, mientras que en la circulación sistémica, especialmente en el lecho esplácnico, los vasos están congestivos y con un flujo aumentado. Por lo tanto un incremento en las resistencias intrahepáticas asociado a un aumento del flujo venoso portal, mediado a través de la dilatación esplácnica, contribuyen al desarrollo de la hipertensión portal. La consecuencia del flujo y la presión transmural elevada a través de los vasos colaterales a partir de una vasculatura portal hipertensa hacia la circulación venosa sistémica con menor presión, conlleva a muchas de las complicaciones observadas en la hipertensión portal, como la hemorragia por várices esofágicas. La importancia del origen vascular primario de la hipertensión portal se basa en la utilidad de terapias actuales orientadas a revertir estas alteraciones hemodinámicas, como los nitratos que reducen la presión portal, a través de vasodilatación intrahepática directa y los P bloqueadores y octreótida, que reducen la vasodilatación esplácnica y el flujo venoso portal. Además, existen nuevas evidencias en relación con los mecanismos moleculares de vías de señalización contráctil de las células estelares hepáticas y complejas vías de regulación de sustancias vasoactivas en las células endoteliales hepáticas que han ayudado a entender mejor estos procesos esenciales para el desarrollo de terapias experimentales para la hipertensión portal. Este artículo revisa los conceptos actuales relacionados con los mecanismos celulares causales de las alteraciones hemodinámicas que caracterizan y condicionan el desarrollo de la hipertensión portal.

Bovine dialyzable leukocyte extract protects against LPS-induced, murine endotoxic shock.

The pathophysiology of endotoxic shock is characterized by the activation of multiple pro-inflammatory genes and their products which initiate the inflammatory process. Endotoxic shock is a serious condition with high mortality. Bovine dialyzable leukocyte extract (bDLE) is a dialyzate of a heterogeneous mixture of low molecular weight substances released from disintegrated leukocytes of the blood or lymphoid tissue obtained from homogenized bovine spleen. bDLE is clinically effective for a broad spectrum of diseases. To determine whether bDLE improves survival and modulates the expression of pro-inflammatory cytokine genes in LPS-induced, murine endotoxic shock, Balb/C mice were treated with bDLE (1 U) after pretreatment with LPS (17 mg/kg). The bDLE improved survival (90%), suppressed IL-10 and IL-6, and decreased IL-1beta, TNF-alpha, and IL-12p40 mRNA expression; and decreased the production of IL-10 (P<0.01), TNF-alpha (P<0.01), and IL-6 (P<0.01) in LPS-induced, murine endotoxic shock. Our results demonstrate that bDLE leads to improved survival in LPS-induced endotoxic shock in mice, modulating the pro-inflammatory cytokine gene expression, suggesting that bDLE is an effective therapeutic agent for inflammatory illnesses associated with an unbalanced expression of pro-inflammatory cytokine genes such as in endotoxic shock, rheumatic arthritis and other diseases.

Exposure levels of asthmatic children to allergens, endotoxins, and serine proteases in a tropical environment.

A cross-sectional study was conducted in Bayamón, Puerto Rico, to identify and quantify indoor allergens, serine proteases, and bacterial endotoxin present in homes of asthmatic children. A total of 126 dust samples from houses were obtained from the entire mattress and bedside floor. Most of the patients had detectable levels of mite, cockroach, cat, and dog allergens. Mold allergens were found only in bedside floor dust samples. Mouse allergens were not detected. Forty-two percent, 36.5%, and 1.8% of the patients demonstrated exposures to sensitizing levels of mite, Bla g 1 and cat allergens, respectively. The percentage of patients exposed to high levels of allergens capable of triggering asthma symptoms was 33.3% and 26.4% for mite and Bla g 1 allergens. Only dog allergen, bacterial endotoxin, elastase, and trypsin were associated with asthma symptoms. Eighty-nine percent of the asthmatic children were exposed to endotoxin concentrations greater than 100 EU/mg dust, and more than half of the patients were exposed to high levels of serine proteases. Our study indicates that indoor concentrations of allergens traditionally associated with asthma symptoms and severity may not be applicable in tropical environments and highly ventilated households. In fact, in the study population, endotoxins, dog allergen, and serine proteases may play a dominant role in the induction of asthma symptoms.

Radiographic evaluation of the effect of endotoxin (LPS) plus calcium hydroxide on apical and periapical tissues of dogs.

The aim of this study was the radiographic evaluation of the apical and periapical region of dog teeth submitted to intracanal bacterial endotoxin (lipopolysaccharide, LPS), associated or not with calcium hydroxide. After removal of the pulp, 60 premolars were divided into four groups and were filled with bacterial endotoxin (group 1), bacterial endotoxin plus calcium hydroxide (group 2), saline solution (group 3), or periapical lesions were induced with no treatment (group 4), for a period of 30 days. Similar periapical lesions were observed in groups 1 and 4. The lamina dura was intact in groups 2 and 3. Bacterial endotoxin (LPS) caused radiographically visible periapical lesions, but when associated with calcium hydroxide, this endotoxin was detoxified.