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Recent work shows that at least two cycles of antigen challenge applied in a 7-day interval are required to yield tissue eosinophil accumulation in IgE-passively sensitized rats. Since interleukin (IL)-13 is widely regarded as a key mediator in eosinophilic responses associated with mast cells and IgE, we investigated whether this cytokine could replace the first cycle of sensitization and challenge in its proeosinophilic role. We found that IL-13 (25 and 50 ng/cavity) injected into the rat pleural space led to eotaxin generation and a dose-dependent accumulation of eosinophils following IgE-passive sensitization and challenge 7 days later. IL-13 failed to cause eosinophil chemotaxis in vitro but induced eosinophil accumulation into the pleural cavity of naïve rats, which peaked 1 day and faded 72 h post-challenge. No changes were found 1 week after intrapleural injection of IL-13, except an approximately 40-50% increase in the number of adhered and non-adhered pleural mast cells. As recovered from the pleural effluent 1 week after IL-13, mast cells expressed the same amount of IgE bound on their surface as compared to controls. However, they generated 3-fold more LTC(4) following IgE-sensitization and challenge in vitro, keeping intact the amount of histamine released. Finally, pretreatment with zileuton (50 microg/cavity) 1 h before allergen challenge prevented eosinophil accumulation in those animals injected with IL-13 1 week before. In conclusion, our findings show that IL-13 causes a long-term exacerbation of the IgE-mediated eosinophilic response in a mechanism associated with heightened cysteinyl-leukotriene (cys-LT) production by resident mast cells.

The prevalence of atopic diseases and diabetes is increasing worldwide though the concurrence of these pathologies in individual patients is found less frequent than it would be predicted. Moreover, co-existence of diabetes and allergy is generally marked by attenuation of their respective symptoms, and effective treatment of one disease exacerbates the other. This review gives an update of the state-of-the-art concerning the intercurrence of allergy and diabetes, particularly focusing on the consequences to the allergen-evoked vascular and cellular changes. It is proposed that the reduction in mast cell numbers and reactivity may be a pivotal mechanism behind the mutual exclusion phenomenon.

We previously reported that alloxan-induced diabetes results in reduction in the number and reactivity of mast cells at different body sites. In this study, the influence of diabetes on thymic mast cells was investigated. Thymuses from diabetic rats showed marked alterations including shrinkage, thymocyte depletion, and increase in the extracellular matrix network, as compared to those profiles seen in normal animals. Nevertheless, we noted that the number and reactivity of mast cells remained unchanged. These findings indicate that although diabetes leads to critical alterations in the thymus, the local mast cell population is refractory to its effect. This suggests that thymic mast cells are under a different regulation as compared to those located in other tissues.

In this study, we investigated the influence of intracellular cyclic adenosine monophosphate (cAMP) changes on the rat mast cell hyporesponsiveness following immunological and non-immunological stimuli. Compared with mast cells from normal rats, those recovered from 21-day diabetic animals showed a significant augmentation in the intracellular levels of cAMP, in directly correlated with secretion of lower amounts of histamine after stimulation with antigen, bradykinin and compound 48/80 in vitro. Incubation of normal mast cells with selective inhibitors of phosphodiesterase type 4 (PDE 4) rolipram, NCS 613 and RP 73401, or the cell permeable analogue N6-2'-O-dibutyryladenosine 3':5'-cyclic monophosphate (db cAMP), led to a decrease of histamine secretion in vitro. However, the effectiveness of either NCS 613 or db cAMP in inhibiting antigen-induced degranulation is comparable in both normal and diabetic mast cells. We suggest that (a) there is a close correlation between higher levels of intracellular cAMP and hyporesponsiveness of diabetic mast cells, phenomena probably associated with a reduction in the expression and/or activity of PDE 4 and that (b) the mechanism of cAMP-mediated down-regulation of mast cell function is saturated in diabetic rats.

Human abdominal angiostrongyliasis is a severe eosinophilic disease caused by Angiostrongylus costaricensis. Previous studies have demonstrated that wild rodents are critically involved as definitive hosts to this nematode in nature. In this study, we have evaluated the susceptibility of Wistar rats (Rattus norvegicus) to A. costaricensis infection. Kinetics of parasitological and pathological changes, including the number of adult worms recovered from mesenteric arteries, and of IgE, mast cell and eosinophil levels in several compartments have been assessed. The oral inoculation of third-stage larvae (L3) into adult Wistar rats led to a marked accumulation of worms in the branches of the mesenteric arteries 25 and 50 days post-inoculation. Intense bone marrow eosinophilia ranging from 7 to 50 days was accompanied by marked accumulation of eosinophils in the blood, peritoneal and bronchoalveolar spaces. Eosinophilic periarteritis, oedema and granuloma in the intestinal and lung tissues were also histologically evident. Total serum IgE and specific anti-parasite IgE peaked at 25 days post-infection, as measured by ELISA and by the passive cutaneous anaphylaxis test, respectively. At that time point, there was a drastic reduction in the number of intact mast cells in the peritoneal effluent. These findings indicate that Wistar rats are permissive to A. costaricensis infection. IgE-mast cell activation and massive tissue eosinophil infiltration are marked features in the process and are likely to play a crucial role in the immune-response evoked by this parasite.

Mast cells are pivotal secretory cells primarily implicated in allergen-evoked inflammatory responses and are downregulated following experimental chemical diabetes. Here we tested the hypothesis that a decrease in the number and reactivity of mast cells would account for the hyporesponsiveness of diabetic rats to allergen-induced inflammation. We found that the anaphylactic release of histamine from sensitized ileum, trachea and skin tissues was clearly reduced in rats turned diabetic via intravenous administration of alloxan. Likewise, actively and passively sensitized diabetic rats mounted a weaker allergen-evoked pleural mast cell degranulation and protein extravasation, as compared to sensitized nondiabetic animals, which paralleled a marked reduction in the mast cell population in the pleural cavity. The number of mast cells enumerated in the mesentery from diabetic rats was also clearly reduced. The allergen-evoked plasma leakage in diabetic rats was restored by the transfer of mast cells from sensitized nondiabetic rats. Moreover, the adoptive transfer of sensitized mast cells from diabetics to naive animals led to a lower allergen-induced exudation as compared to the response noted after the transfer of sensitized naive mast cells. Purified mast cells from diabetic rats were hyporesponsive to antigen and compound 48/80 stimulation in vitro as attested by histamine release. Thus, our results show that the phenomenon of refractoriness of diabetic animals to allergen challenge appears to be accounted for by a reduction in the number and reactivity of mast cells.

During the past year, the EMBnet Executive Board (EB) had regular monthly meetings either via Skype or using Adobe Connect. These meetings were always carried out with the Interim Board (IB) composed by Teresa K. Attwood and Etienne de Villiers. The IB was indeed established during the Annual General Meeting 2015 to support the new EB to move its first steps forward the AGM 2016. The EB regularly invited also Special Interest Groups (SIGs) Chairs to participate at EB-IB meetings. Additional monthly meetings open to the full EMBnet constituency were also convened. In this report, we provide a brief overview of facts, activities and results of this last year that span from June 2015 to October 2016.

Monoterpenes, compounds mainly presented in essential oils, have important pharmacological actions. Isopropoxy-carvacrol (IPC) is a derivative of the monoterpene carvacrol, and its pharmacological properties have not yet been investigated. The aim of this study was to analyse the acute anti-inflammatory and antinociceptive properties of IPC. Mice (25–30 g) and rats (150–230 g) were pre-treated (i.p.) with IPC at the doses of 10, 30 or 100 mg/kg or vehicle (Tween 80, 0.5%), 30 min. before injection of the phlogistic agents. Both the first and the second phases of formalin-induced nociception were significantly reduced by IPC (100 mg/kg). Injection of carrageenan in mice paw reduced the threshold of stimulus intensity, applied with an analgesymeter, necessary to cause paw withdrawal, which was significantly reduced by 100 mg/kg of IPC. The area under curve (0–4 hr) of rat paw oedema induced by injection of carrageenan was also significantly diminished by the administration of IPC (100 mg/kg). Administration of 12-O-tetradecanoylphorbol-13-acetate (TPA) markedly increased mice ear oedema and myeloperidase (MPO) activity. Topical co-administration of IPC (0.3–3 mg/ear) during the induction did not affect TPA-induced ear oedema, but significantly decreased MPO activity in the ears, when compared with the vehicle. In in vitro experiments, IPC reduced lipoperoxidation induced by different stimuli, showed nitric oxide scavenger activity and did not interfere with murine macrophage viability in concentrations up to 100 μg/mL. These results demonstrate that IPC exerts acute anti-inflammatory and antinociceptive activities, suggesting that it may represent an alternative in the development of new future therapeutic strategies.

Background Essential oils are complex systems which consist mainly of volatile compounds of lipophilic source, as terpenes, sesquiterpenes and some non-terpenes [1]. Resulting from secondary metabolism, essential oil from Croton argyrophyllus (EOCA) is commonly extracted from its leaves. It is known that the essential oils from Croton species exhibit good antioxidant activity against DPPH free radical and reactive oxygen species (ROS) [2]. Several diseases are attributed to the increase of free radicals that show the importance of endogenous antioxidants like enzyme superoxide dismutase (SOD). Considering the use in folk medicine, it is relevant to find which concentration of EOCA has antioxidant effect against free radicals. To answer this question, the study aimed to evaluate ROS production in peritoneal macrophages of essential oil from C. argyrophyllus.

In the present study, siaresinolic acid (siaresinol, SA) was isolated from the leaves of Sabicea grisea and studied to evaluate its antinociceptive and anti-inflammatory activity. The antinociceptive effect of SA was investigated in mice using different animal models to study pain. In the acetic acid-induced writhing test, intraperitoneal (i.p.) injection of SA (0.1, 1, and 10 mg/kg, i.p.) 1 h before a pain stimulus significantly reduced the nociceptive response (by 42.3, 68.2, and 70.9 %, respectively). Pretreatment with glibenclamide, but not with yohimbine, metoclopramide, ketanserin, or naloxone, restored the antinociceptive effect induced by SA in the writhing test, suggesting that the K(+)ATP channel pathway might be involved in its mechanism of action. In the formalin test, SA (1 mg/kg, i.p.) decreased licking time in the second phase only, thereby indicating an anti-inflammatory effect. In the hot plate test, there was no significant difference in nociceptive behavior. In the rota-rod test, it was verified that a high dose of SA (10 mg/kg, i.p.) did not affect the locomotor activity of mice. In the pleurisy model, induced by carrageenan, treatment with SA inhibited important events involved in inflammatory responses, namely leukocyte influx, plasma leakage, and increased inflammatory mediators (TNF-α, IL-1β, and chemokine CXCL1), in the pleural exudate. Additionally, SA itself was not cytotoxic when evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in macrophages cultured for 24 h at concentrations ranging from 1 to 200 μg/mL. These results suggest, for the first time, that SA attenuates nociceptive behavior through mechanisms involving receptors for ATP-dependent potassium channels, in addition to suppressing acute inflammatory responses.

Titulo en espanol: Analisis genomico del resistoma de la cepa de Acinetobacter baumannii ABIBUN 10 7m multi-resistente y persistente en hospitales colombianos Titulo en ingles: Genomic analysis of the resistoma of the strain of Acinetobacter baumannii ABIBUN 107m multi-resistant and persistent in Colombian hospitals Titulo corto: Analisis genomico del resistoma de la cepa de Acinetobacter baumannii Resumen: Acinetobacter baumannii es una bacteria, causante de infecciones asociadas a la atencion en salud como neumonia, septicemia, meningitis e infecciones urinarias entre otras. Se caracteriza por su capacidad para desarrollar y acumular rapidamente una gran variedad de mecanismos de resistencia a antibioticos. En esta investigacion se realizo el analisis genomico de una cepa de A. baumannii ABIBUN 107m que forma parte de un clon persistente en hospitales colombianos, resistente a los antibioticos carbapenemicos (imipenem y meropenem), antibioticos de eleccion en el tratamiento infecciones causadas por este microorganismo. El genoma de esta bacteria fue secuenciado utilizando tecnicas de alto rendimiento, ensamblado y anotado, obteniendose un genoma constituido por 3954000 pb con 56 c ontigs; consta de 4256 genes con un tamano promedio de 912 pb; 3796 CDS de los cuales por anotacion 2884 se asignaron a COG; 57 tRNA y un porcentaje de GC de 38,74%. A. baumannii ABIBUN 107m es resistente a b-lactamicos, aminoglicosidos, quinolonas, tetraciclina, sulfonamida y colistina. En su genoma se localizaron genes asociados con el perfil de resistencia ya que presenta serin b-lactamasas ( bla ADC-38 , bla OXA-64 , bla OXA-23 , bla amp C-like , bla amp(H) -like ), metalo b-lactamasa_B; proteinas de union a penicilina de elevada masa molecular, secuencias de insercion tipo ISAba1; mutaciones en los genes de DNA girasa y topoisomerasa IV subunidad A ( gyr A y par C); enzimas modificadoras de aminoglicosidos ( aph A -like , aad A- like ); cloranfenicol aciltransferasa ( cat ) y dehidropteroato sintasa ( sul-1) . Se identificaron genes pertenecientes a cinco familias de sistemas de eflujo (RND, MATE, MSF, ATP, SMR). Abstract: Acinetobacter baumannii is a bacterium causing health care associated infections such as pneumonia, septicemia, meningitis and urinary infections amongst others. It has great capacity to quickly develop and gather a big variety of drug resistance mechanisms. In this research, the genome of strain A. baumannii ABIBUN 107m was analyzed wich forms part of a persistent clon in Colombian hospitals and it’s also resistant to carbapenems (imipenem and meropenem), which are the election antibiotics for treatment of infections caused by this microorganism. The genome was sequenced using high performance technology, assembled and annotated. As a result, we obtained a 3954000 bp genome, with 56 c ontigs; 4256 genes with average size of 912 bp; 3796 CDS; 2884 were assigned to COG; 57 tRNA and GC percentage of 38,74%. The A. baumannii strain ABIBUN 107m, is resistant to the following antibiotic groups: b-lactams, aminoglycosides, quinolones, tetracycline, sulfonamide and colistin. Genes associated with this resistance profile were found in A. baumannii ABIBUN 107m genome serino b-lactamases ( bla ADC-38 , bla OXA-64 , bla OXA-23 , bla amp C-like , bla amp(H) -like , metallo b-lactamase_B; High Molecular Mass penicillin binding proteins, IS Aba 1 type insertion sequences, mutations of DNA gyrase and topoisomerase IV subunit A ( gyr A and par C); aminoglycoside modifying enzymes ( aph A -like , aad A- like ); choramphenicol acyltransferase (cat) and dehydropteroate synthase ( sul -1). Genes belonging to five different efflux systems were identified (RND, MATE, MSF, ATP, SMR). Key words: Acinetobacter baumannnii MDR , resistome, Colombia Recibido: febrero 26 de 2014 Aprobado: octubre 17 de 2014

This study aimed at synthesizing the carvacrol propionate (CP) and evaluating its pharmacological profile. CP was obtained from carvacrol and propionyl chloride through an esterification reaction. Male Swiss mice were treated with CP (25, 50, or 100 mg/kg). We evaluated the analgesic effect, mechanical hyperalgesia, and anti-inflammatory effect. Pre-treatment with CP inhibited (p

Interactions between cells and microenvironments are essential to cellular functions such as survival, exocytosis and differentiation. Cell adhesion to the extracellular matrix (ECM) evokes a variety of biophysical changes in cellular organization, including modification of the cytoskeleton and plasma membrane. In fact, the cytoskeleton and plasma membrane are structures that mediate adherent contacts with the ECM; therefore, they are closely correlated. Considering that the mechanical properties of the cell could be affected by cell adhesion-induced changes in the cytoskeleton, the purpose of this study was to investigate the influence of the ECM on the elastic properties of fixed macrophage cells using atomic force microscopy. The results showed that there was an increase (~50 %) in the Young’s modulus of macrophages adhered to an ECM-coated substrate as compared with an uncoated glass substrate. In addition, cytochalasin D-treated cells had a 1.8-fold reduction of the Young’s modulus of the cells, indicating the contribution of the actin cytoskeleton to the elastic properties of the cell. Our findings show that cell adhesion influences the mechanical properties of the plasma membrane, providing new information toward understanding the influence of the ECM on elastic alterations of macrophage cell membranes.

Beta-Lactamases OXA and CTX-M are highly disseminated and confer resistance to antibiotics. This project gathers, classifies and analyzes information from indexed articles and databases as UNIPROT, EMBL and PDB, in order to correlate molecular data with activity profile of beta-lactamases against beta-Lactamic antibiotics. Sequences of CTX-M and OXA enzymes are analyzed by ClustalW multiple alignment and distance trees are built by Neighbor – joining with default parameters and a Bootstrap of 1000, through MEGA 5.0.

Nowadays a widely production and low cost of sequencing has allowed the extension of metagenomics in order to explore the genomic information from diverse environments. This offers the opportunity to examine new approaches for sequence binning and functional assignment. Driving of metagenomic studies using high throughput sequencing, usually follows the same pipeline used to analyze single genomes: sequence assembling, gene prediction, functional annotation and phylogenetic classification of reads, contigs or scaffolds, nevertheless, the accuracy of this approach is limited by the length of the reads or resulting contigs.

This study was undertaken to investigate the possible contribution of the blockade of eotaxin generation to the anti-eosinophilotactic effect of phosphodiesterase (PDE) type 4 inhibitors. In some experiments, the putative synergistic interaction between PDE type 4 inhibitors and the β2-agonist salbutamol was also assessed. Sensitized guinea-pigs aerosolized with antigen (5% ovalbumin, OVA) responded with a significant increase in eotaxin and eosinophil levels in the bronchoalveolar lavage fluid (BALF) at 6 h. Eosinophil recruitment was inhibited by both PDE type 4 inhibitors rolipram (5 mg kg−1, i.p.) and RP 73401 (5 mg kg−1, i.p.) treatments. In contrast, only rolipram inhibited eotaxin production. Sensitized rats intrapleurally challenged (i.pl.) with antigen (OVA, 12 μg cavity−1) showed a marked eosinophil infiltration at 24 h, preceded by eotaxin generation at 6 h. Intravenous administration of a rabbit anti-mouse eotaxin antibody (0.5 mg kg−1) significantly reduced allergen-evoked eosinophilia in this model. Local pretreatment with rolipram (40 μg cavity−1) or RP 73401 (40 μg cavity−1) 1 h before challenge reduced eosinophil accumulation evaluated in the rat pleural effluent, but only the former was active against eotaxin generation. The inhibitors of PDE type 3 (SK&F 94836) and type 5 (zaprinast) failed to alter allergen-evoked eosinophil recruitment in rats. Local injection of β2-agonist salbutamol (20 μg cavity−1) inhibited both eosinophil accumulation and eotaxin production following pleurisy. The former was better inhibited when salbutamol and rolipram were administered in combination. Treatment with rolipram and RP 73401 dose-dependently inhibited eosinophil adhesion and migration in vitro. These effects were clearly potentiated by salbutamol at concentrations that had no effect alone. Our findings indicate that although rolipram and RP 73401 are equally effective in inhibiting allergen-induced eosinophil infiltration only the former prevents eotaxin formation, indicating that PDE 4 inhibitors impair eosinophil accumulation by mechanisms independent of eotaxin production blockade. British Journal of Pharmacology (2001) 134, 283–294; doi:10.1038/sj.bjp.0704233

Rats turned diabetic by treatment with alloxan exhibit a significant reduction in serosal mast cell numbersin parallel with decreased insulin levels in the plasma. Our aim was to investigate the putative involvement of endogenous glucocorticoid hormone in this phenomenon. The findings indicated that rats treated with alloxan responded with an increase in levels of serum corticosterone concomitantly with decreased mast cell numbers in the pleural space. We found that either surgical bilateral adrenalectomy or pretreatment with the steroid antagonist RU 486 (20 mg/kg, i.p.) impaired the drop in pleural mast cell counts in alloxinated rats. Administration of insulin (15 U/kg) prevented the increase in corticosterone levels and restored pleural mast cell levels in diabetic animals. In addition, treatment of naive rats with corticosterone (0.5 mg/kg, s.c.) or dexamethasone (0.1 mg/kg, s.c.), for 3 consecutive days, led to a reduction in the number of mast cells recovered from the pleural cavity as noted in diabetic animals. In contrast, insulin reduced serum corticosterone levels and induced a significant elevation in pleural mast cell numbers in naive rats. We conclude that there is a causative relationship between increased levels of glucocorticoids and down-regulation of mast cell numbers associated with the diabetic state, both phenomena clearly sensitive to insulin.

Objective: To investigate the involvement of the fibrinogen-fibrin system in the acute reduction of the resident leukocyte population following pleural inflammation.¶Methods: Sensitized and naive rats were injected intrapleurally (i.pl.) with antigen (ovalbumin) and platelet-activating factor (PAF) or bradykinin, respectively. Heparin (0.25 U/ cavity), EDTA (80 μg/cavity) and hirudin (1 U/cavity) were injected locally 5 min before challenge, whereas fucoidin was injected intraperitoneally 30 min before stimulation.¶Results: Antigen challenge led to a rapid reduction in the number of resident leukocytes 30 min post-challenge (from 7.7 ± 0.4 × 106 cells/cavity to 2.3 ± 0.2 × 106 cells/cavity, n = 6, p < 0.001). The pleural stimulation of naive rats with either PAF or bradykinin also led to a significant decrease in the pleural leukocyte population, which occurred in parallel with the formation of a fib rin meshwork containing captured cells, as attested by electron microscopy. Heparin prevented the drop in the total leukocyte numbers, without modifying either plasma leakage or histamine release at 30 min or the subsequent neutrophil and eosinophil infiltration noted 4 and 24h post-challenge, respectively. Similarly, hirudin and EDTA prevented the antigen-induced leukocyte disappearance reaction. Heparin also impaired the drop in the pleural leukocyte numbers evoked by PAF and bradykinin.¶Conclusion: Our data show that the pleural resident cell disappearance phenomenon noted early after inflammatory provocation depends on the activation of the fibrinogen-fibrin system, and is not required for the subsequent leukocyte recruitment.

The draft genome sequences of the strains Acinetobacter baumannii 107m, Acinetobacter nosocomialis 28F, and Acinetobacter pittii 42F, isolated from Colombian hospitals, are reported here. These isolates are causative of nosocomial infections and are classified as multidrug resistant, as they showed resistance to four different antibiotic groups.

The decoction of the stem barks from Bowdichia virgilioides KUNTH is a folk remedy used to treat inflammatory disorders in Latin American and Brazil. In the present study, the wound healing activity of aqueous extract of the stem bark from B. virgilioides, called AEBv, was evaluated by the rate of healing by wound contraction and period of epithelization at different days post-wound using the wound excisional model. On day 9, the AEBv-treated animals exhibited significative reduction in the wound area when compared with controls. In wound infected with S. aureus, the AEBv significantly improved the wound contraction when compared to the saline-treated mice. The histological analysis showed that AEBv induced a collagen deposition, increase in the fibroblast count and few inflammatory cells than compared to saline-treated group. The expression of collagen type I was increased in the group treated with AEBv as indicated by immunohistochemical staining. In vitro, the AEBv was effective only against S. aureus but not against P. aeruginosa. Together, the results of this study demonstrate, for the first time, the healing and antimicrobiological effects of aqueous extract of the stem bark from B. virgilioides in the therapy of skin wounds. A decocção das cascas do caule de Bowdichia virgilioides Kunth é um medicamento popular usado para tratar doenças inflamatórias na América Latina e no Brasil. Neste estudo, a atividade de cicatrização de feridas do extrato aquoso da casca do caule de B. virgilioides, chamado AEBv, foi avaliada pela contração da ferida e pelo período de epitelização em diferentes dias pós-ferida usando o modelo ferida excisional. No nono dia, os animais tratados com AEBv apresentaram uma redução significativa na área da ferida, quando comparados com os controles. Nas feridas infectadas com S. aureus, o AEBv melhorou significativamente a contração da ferida quando comparado com os camundongos tratados com solução salina. A análise histológica mostrou que AEBv induziu uma deposição de colágeno, aumento na contagem de fibroblastos e poucas células inflamatórias do que em relação ao grupo tratado com solução salina. A expressão de colágeno tipo I mostrou-se aumentada no grupo tratado com AEBv como indicado pela coloração imuno histoquímica. In vitro, o AEBv foi eficaz apenas contra S. aureus, mas não contra P. aeruginosa. Juntos, os resultados deste estudo demonstram, pela primeira vez, a cura e os efeitos antimicrobianos do extrato aquoso da casca do caule de B. virgilioides na terapia de feridas cutâneas.

Reflux laryngitis is a common clinic complication of nasogastric intubation (NSGI). Since there is no report concerning the effects of low level laser therapy (LLLT) on reflux laryngitis, this study aimed to analyze the protective effect of single and combined therapies with low level laser at the doses of 2.1J and 2.1+1.2 J with a total irradiation time of 30s and 30+30 s, respectively, on a model of neurogenic reflux laryngitis. NSGI was performed in Wistar rats, assigned into groups: NGI (no treatment), NLT17.5 (single therapy), and NLT17.5/10.0 (combined therapy, applied sequentially). Additional non-intubated and non-irradiated rats were use as controls (CTR). Myeloperoxidase (MPO) activity was assessed by colorimetric method after the intubation period (on days 1, 3, 5, and 7), whereas paraffin-embedded laryngeal specimens were used to carry out histopathological analysis of the inflammatory response, granulation tissue, and collagen deposition 7 days after NSGI. Significant reduction in MPO activity (p0.05) and in the severity of the inflammatory response (p0.05), and improvement in the granulation tissue (p0.05) was observed in NLT17.5/10.0 group. Mast cells count was significantly decreased in NGI and NLT17.5 groups (p0.001), whereas no difference was observed between NLT17.5/10.0 and CTR groups (p0.05). NLT17.5/10.0 group also showed better collagenization pattern, in comparison to NGI and NLT17.5 groups. This study suggests that the combined therapy successfully modulated the inflammatory response and collagenization in experimental model of NSGI-induced neurogenic laryngitis.

Biologists work constantly to find answers that allow proposing solutions to problems that affect everyday life. Computing has the necessary elements to support the work of biologists using bioinformatics. The characterization of soils for creole potato cultivation in Colombia demands interdisciplinary work between biology and computer science. In biology, the hybridization of chains of DNA allows to carry out the soil characterization as a laboratory activity. The computer science allows recreating this process of hybridization with the help of appropriate computer equipment, in order to reproduce the process of computational hybridization of sequences of DNA. This process is known as hybridization in silico. A general problem in biological information processing is the high response time. This paper proposes a parallelized version of hybridizer in silico system that runs on a computer cluster using mpiBLAST implementation. We propose an algorithm called Hybridizer In silico for Paralleling Processing - HISPeP-UN, which aims to increase the performance and to reduce system response time, allowing characterization results, in this case of soil, at reasonable times.

Introduction: The different ways for selecting single nucleotide polymorphisms have been related to paradoxical conclusions about their usefulness in predicting chronic fatigue syndrome even when using the same dataset. Objective: To evaluate the efficacy in predicting this syndrome by using polymorphisms selected by a supervised approach that is claimed to be a method that helps identifying their optimal profile. Materials and methods: We eliminated those polymorphisms that did not meet the Hardy-Weinberg equilibrium. Next, the profile of polymorphisms was obtained through the supervised approach and three aspects were evaluated: comparison of prediction accuracy with the accuracy of a profile that was based on linkage disequilibrium, assessment of the efficacy in determining a higher risk stratum, and estimating the algorithm influence on accuracy. Results: A valid profile (p

α-Terpineol (TPN), a volatile monoterpene alcohol, is relatively non-toxic and one of the major components of the essential oils of various plant species. In this study, we tested for the antihypernociceptive activity of TPN (25, 50 or 100 mg/kg, i.p.) in mice using mechanical models of hypernociception induced by carrageenan (CG, 300 μg/paw) and the involvement of important mediators of its cascade signalling, such as tumour necrosis factor-α (TNF-α, 100 pg/paw), prostaglandin E₂ (PGE₂, 100 ng/paw) or dopamine (DA, 30 μg/paw). We also investigated the anti-inflammatory effect of TPN on the model of carrageenan-induced pleurisy and the LPS-induced nitrite production in murine macrophages. Pre-systemic treatment with TPN (25, 50 or 100 mg/kg, i.p.) inhibited the development of mechanical hypernociception induced by CG or TNF-α. A similar effect was also observed upon PGE₂ and DA administration. In addition, TPN significantly inhibited the neutrophil influx in the pleurisy model. TPN (1, 10 and 100 μg/mL) also significantly reduced (p < 0.01) nitrite production in vitro. Our results provide information about the antinociceptive and anti-inflammatory properties of TPN on mechanical hypernociception and suggest that this compound might be potentially interesting in the development of new clinically relevant drugs for the management of painful and/or inflammatory disease.

The present study was carried out to investigate the anti-inflammatory effect of the hexane extract of the leaves from Clusia nemorosa G. Mey, called HECn, using carrageenan-induced mice pleurisy and cotton pellet-induced mice granuloma. Additionally, the ability of HECn to affect both neutrophil migration as viability was investigated by use of the Boyden chamber assay and flow cytometry, respectively. The HECn significantly inhibited exudation, total leukocytes and neutrophils influx, as well as TNFα levels in carrageenan-induced pleurisy. However, the extract not suppressed the granulomatous tissue formation in the cotton pellet-induced granuloma test. Experiments performed in vitro revealed that HECn on human neutrophils inhibited a dose-dependent manner the CXCL1-induced neutrophil chemotaxis. Furthermore, HECn also inhibited the chemoattraction of human neutrophils induced by formyl-methionyl-leucyl-phenylalanine (fMLP), leukotriene B4 (LTB4) and platelet activating factor (PAF) in a Boyden chamber. However, this same treatment not was able to induce apoptosis. The results obtained in this study showed that the extract from leaves of C. nemorosa possess a potent inhibitory activity in acute model of inflammation, being the effects mediated, in part, by inhibition of neutrophil responsiveness. These results indicate that C. nemorosa could be a good source for anti-inflammatory compounds.

Pre-mRNA splicing is an essential step in the post-transcriptional gene expression control involving protein-splicing factors like U2AF, which is exported to the cytoplasm and implicated in additional cellular functions. Identification of U2AF-associated mRNAs under native conditions was performed by immunoprecipitation and hybridization to Affymetrix GeneChip. Normalization and gene selection methods were performed, but the results were not reliable as they were different for different procedures, mainly because more than 20% of the mRNAs detected are differently enriched and the common normalization methods are based on small differences between them. We implemented a background correction method inspired in a non-specific hybridization method used for pre-processing data from ChIP-Chip technology. In this work, linear regression models are used to model in each array the non-specific hybridization, accounting for interactions between each three consecutive nucleotides into the probe sequence. Every probe intensity on the array was standardized using its predicted intensity and the probes' variance for similar predicted intensities. The standardized probe intensity values showed no need for further normalization and could be directly compared. We propose a probe set score, and a probe set enrichment value (ENRval) and its respective p-value for gene enrichment selection.

Introduction. The different selection of single nucleotide polymorphisms has been related with paradoxical conclusions about their usefulness to predict the chronic fatigue syndrome, even by using the same dataset.Objective. To evaluate the efficacy to predict this syndrome of polymorphisms selected by a supervised approach that is claimed as a method that helps to identify the optimal profile of polymorphisms.Materials and methods. From the 44 polymorphisms, we eliminated those out of Hardy-Weinberg equilibrium. Next, we obtained the profile of polymorphisms by means of the supervised approach and evaluated three aspects: comparison of prediction accuracy with the accuracy of the profile that was based on linkage disequilibrium, assessment of the efficacy to determine a higher risk stratum, and estimation of the algorithm influence on prediction accuracy.Results. We obtained a valid profile (pConclusions. The supervised approach allowed discovering a valid and reliable profile of polymorphisms associated with chronic fatigue syndrome. It showed the highest reported accuracy in this dataset that was increased when it was combined with clinical data.Introducción. La diferente selección de polimorfismos de nucleótido único se ha relacionado con conclusiones paradójicas respecto a su utilidad para predecir el síndrome de fatiga crónica, incluso utilizando los mismos datos.Objetivo. Evaluar la eficacia para predecir este síndrome de los polimorfismos seleccionados mediante un enfoque supervisado, método que ofrece ayudar a identificar el perfil óptimo de polimorfismos.Materiales y métodos. De 44 polimorfismos eliminamos los que no estaban en equilibrio de Hardy-Weinberg. Luego obtuvimos el perfil de polimorfismos mediante el enfoque supervisado y evaluamos tres aspectos: comparación de la exactitud de predicción con la del perfil obtenido mediante una selección basada en desequilibrio de ligamiento, evaluación de la eficacia para determinar un estrato con mayor riesgo y estimación de la influencia del algoritmo de clasificación sobre la exactitud de predicción.Resultados. Obtuvimos un perfil válido (pConclusiones. El enfoque supervisado permitió descubrir un perfil válido y confiable de polimorfismos asociado al síndrome de fatiga crónica. Mostró la mayor exactitud reportada con estos datos que aumentó al combinarse con variables clínicas.

The different ways for selecting single nucleotide polymorphisms have been related to paradoxical conclusions about their usefulness in predicting chronic fatigue syndrome even when using the same dataset.To evaluate the efficacy in predicting this syndrome by using polymorphisms selected by a supervised approach that is claimed to be a method that helps identifying their optimal profile.We eliminated those polymorphisms that did not meet the Hardy-Weinberg equilibrium. Next, the profile of polymorphisms was obtained through the supervised approach and three aspects were evaluated: comparison of prediction accuracy with the accuracy of a profile that was based on linkage disequilibrium, assessment of the efficacy in determining a higher risk stratum, and estimating the algorithm influence on accuracy.A valid profile (p0.01) was obtained with a higher accuracy than the one based on linkage disequilibrium, 72.8 vs. 62.2% (p0.01). This profile included two known polymorphisms associated with chronic fatigue syndrome, the NR3C1_11159943 major allele and the 5HTT_7911132 minor allele. Muscular pain or sinus nasal symptoms in the stratum with the profile predicted V with a higher accuracy than those symptoms in the entire dataset, 87.1 vs. 70.4% (p0.01) and 92.5 vs. 71.8% (p0.01) respectively. The profile led to similar accuracies with different algorithms.The supervised approach made it possible to discover a reliable profile of polymorphisms associated with this syndrome. Using this profile, accuracy for this dataset was the highest reported and it increased when the profile was combined with clinical data.

Mast cell number and reactivity have been shown to be down-regulated under diabetic conditions. This study was undertaken in order to investigate the role of the advanced glycation end products in the reduction of mast cell number and reactivity in diabetic rats. The effect of aminoguanidine on mast cell apoptosis was also evaluated. Diabetes was induced by intravenous injection of alloxan into fasted rats and aminoguanidine was administered after 3 days of diabetes induction, once daily for 18 consecutive days. Mast cell apoptosis and levels of Bax, a pro-apoptotic member of Bcl-2 family, were evaluated by TUNEL and western blot, respectively. Diabetes led to increased levels of fructosamine and AGEs in the plasma, an effect prevented by aminoguanidine. Treatment with aminoguanidine restored mast cell numbers in the pleural cavity and in mesenteric tissue of diabetic rats. Aminoguanidine also significantly reversed the diabetes-induced reduction in histamine release, as measured by fluorescence, following activation with substance P or antigen in vitro. Increased apoptosis and levels of Bax in mast cells from diabetic rats were inhibited by aminoguanidine. In conclusion, our findings showed that aminoguanidine restored the number and reactivity of mast cells in diabetic rats, accompanied by suppression of apoptosis, evidencing that advanced glycation end product formation has a critical role in mast cell behavior of diabetic rats.

ResumenEste artículo es parte de los resultados de una investigaciónpara conocer el proceso de gestión de la integración social de lainvestigación de nueve instituciones de educación superior colombianas. En la investigación se analizaron 21 proyectos deinvestigación de diferentes áreas de conocimiento y tipo de investigaciones (básica, aplicada e I+D+I).Las técnicas e instrumentos utilizados fueron la entrevista semiestructurada o enfocada, el grupo focal y análisis documental. Los resultados muestran que la integración es un proceso complejo que implica transferencia y apropiación y que para que exista unagestión de la integración del conocimiento se necesita no solamenteuna capacidad de gestión sobre los diferentes temas involucradosen la integración (intencionalidad, actores, estructuras organizacionales), sino que adicionalmente existan requerimientos y exigencias muy concretas en las políticas de Estado para el fomento a lainvestigación.AbstractThis article is a part of the results of a research aiming at knowingthe management process of social integration of research in nine Colombianuniversities. In the study, 21 research projects in different areas of knowledge and type of research (basic, applied and I+D+I) were analyzedSemi-structure interviews, focal groups and documentary analysis wereused s techniques and instruments to collect data. Results show that the integration is a complex process which implies transference and appropriation, and in order to exist a management of the integration of the knowledge, it is necessary not only to have the ability to manage the different involved topics in the integration (intentionality, actors, organizational structures), but also the existence of very concrete requirements in the State policies for fostering research.

High-throughput techniques are somewhat restricted in developing countries. However, computational resources have evolved in recent years to become available to the general public, with greater ability to solve intense computational problems at low cost. Therefore, the vast amount of information that is currently being generated and the need for finding the underpinnings of several issues in biology, have been the impetus of the computational biology area in Latin America. Colombia is no exception, as its rich genetic diversity has convened the attention of several institutions, including both governmental and academic departments, to find how, where, and when these resources could be employed to its benefit. In this review, we introduce the efforts being made throughout the country to spread the word and establish a strong network from a mid- and long-term perspective.

Background Phytophthora infestans is a devastating oomycete pathogen of potato production worldwide. This review explores the use of computational models for studying the molecular interactions between P. infestans and one of its hosts, Solanum tuberosum. Modeling and conclusion Deterministic logistics models have been widely used to study pathogenicity mechanisms since the early 1950s, and have focused on processes at higher biological resolution levels. In recent years, owing to the availability of high throughput biological data and computational resources, interest in stochastic modeling of plant-pathogen interactions has grown. Stochastic models better reflect the behavior of biological systems. Most modern approaches to plant pathology modeling require molecular kinetics information. Unfortunately, this information is not available for many plant pathogens, including P. infestans. Boolean formalism has compensated for the lack of kinetics; this is especially the case where comparative genomics, protein-protein interactions and differential gene expression are the most common data resources.

Background Members of the genus Phytophthora are notorious pathogens with world-wide distribution. The most devastating species include P. infestans, P. ramorum and P. sojae. In order to develop molecular methods for routinely characterizing their populations and to gain a better insight into the organization and evolution of their genomes, we used an in silico approach to survey and compare simple sequence repeats (SSRs) in transcript sequences from these three species. We compared the occurrence, relative abundance, relative density and cross-species transferability of the SSRs in these oomycetes. Results The number of SSRs in oomycetes transcribed sequences is low and long SSRs are rare. The in silico transferability of SSRs among the Phytophthora species was analyzed for all sets generated, and primers were selected on the basis of similarity as possible candidates for transferability to other Phytophthora species. Sequences encoding putative pathogenicity factors from all three Phytophthora species were also surveyed for presence of SSRs. However, no correlation between gene function and SSR abundance was observed. The SSR survey results, and the primer pairs designed for all SSRs from the three species, were deposited in a public database. Conclusion In all cases the most common SSRs were trinucleotide repeat units with low repeat numbers. A proportion (7.5%) of primers could be transferred with 90% similarity between at least two species of Phytophthora. This information represents a valuable source of molecular markers for use in population genetics, genetic mapping and strain fingerprinting studies of oomycetes, and illustrates how genomic databases can be exploited to generate data-mining filters for SSRs before experimental validation.

Automatically finding new protein domains is a challenge when using the complete collection of known proteins (i.e., UniProt). By limiting the taxonomic range to class insecta, including two full proteomes (A. gambiae and D. melanogaster), we reduced the size of the search space in the hope of finding taxon-specific domains. The MKDOM2 program (http://prodes.toulouse.inra.fr/prodom/xdom/mkdom2.html) was used to cluster the insect proteins into potential domains that were analyzed manually in a second step. We analyzed 219 potential domains, of which 2 were insect-specific. We show that it is possible to find new domains or to extend known domains using a semi-automated method; however the goal to detect class-specific domains was only partially achieved in the sense that the new domains we found were not all insect-specific domains. The files used as input and the resulting output files, as well as extensive descriptions of the domains, are available as supplementary data from http://bioinf.ibun.unal.edu.co/insecta/.

This study was undertaken to investigate the role of the aldose reductase in the refractoriness of diabetic rats to allergic inflammation. Wistar rats were actively sensitized with a mixture of Al(OH)3 plus ovalbumin and intrapleurally challenged with ovalbumin, 14 days later. Diabetes was induced by intravenous injection of alloxan into fasted rats, 7 days before sensitization, and the aldose reductase inhibitor zopolrestat was administered after 3 days of diabetes induction, once a day during 18 consecutive days. The treatment with zopolrestat restored antigen-induced protein extravazation and mast cell degranulation in the pleural cavity of diabetic sensitized rats. Zopolrestat also significantly reversed the suppression in the increase of total and specific levels of serum immunoglobulin E (IgE) noted in sensitized animals under conditions of diabetes. In addition, we noted that the drop in the pleural mast cell numbers as well as the increase in serum corticosterone levels in diabetic rats were inhibited by the drug. Our findings show that zopolrestat restored the hyporesponsiveness of diabetic rats to antigen provocation, in parallel with impairment of alloxan-induced mast cell depletion and hypercorticolism, indicating that polyol pathway activity seems to play an important role in these phenomena.

Mast cell number and reactivity were shown to be down-regulated under diabetic conditions. Since the balance between globular and filamentous actin plays a pivotal role in the activity of secretory cells, we investigated whether an imbalance in that system could underlie the hyporesponsiveness of mast cells in diabetes. The apoptotic state was also evaluated. By means of rhodamine/phalloidine staining of F-actin, we noted that diabetic mast cells exhibited an increase in fluorescence intensity and reduction in cellular size, when compared with cells from normal animals, in parallel with elevation in the percentage of cells developing apoptosis. The levels of Bax, a pro-apoptotic member of Bcl-2 family, appeared increased at baseline in mast cells from diabetic rats compared with normal cells. These phenomena correlated with reduction in histamine and PGD2 release following antigen challenge in vitro. The steroid antagonist RU 486 abolished the reduction of histamine secretion from diabetic mast cells. We conclude that hyporesponsiveness of mast cells noted in diabetes may be accounted for by reduction in actin filament plasticity, in clear association with the rise in the percentage of cells undergoing apoptosis. In addition, the refractoriness of diabetic mast cells to antigen in vitro seems to be dependent on glucocorticoids.

In this study we examined the ability of the furofuran lignan yangambin to influence the local and systemic responses induced by antigen or PAF in actively sensitized or normal rats. Given intraperitoneally 1h before stimulation, yangambin inhibited the pleural neutrophil and eosinophil infiltration evoked by the i.pl. injection of PAF or antigen into normal or 14 day-sensitized rats whereas plasma exudation evoked by both stimuli was unaffected. The pleural neutrophil influx (6h) after LTB 4 stimulation was also significantly inhibited by yangambin. We also evidenced that the hemoconcentration, thrombocytopenia, and leucocytosis noted after i.v. PAF were all attenuated by yangambin. In actively sensitized rats, pretreatment with yangambin failed to modify the antigen-induced hemoconcentration and leucocytosis, but dose-dependently abrogated the thrombocytopenia noted 1h post-stimulation. In vitro, the anaphylactic contraction of longitudinal jejunal segments to antigen challenge was significantly inhibited by yangambin (10 -5 -10 -4 M). Likewise, the contraction of jejunal segments from normal rats to PAF was markedly blocked by yangambin under conditions where the response to 5-hydroxytryptamine (5-HT) was not altered. In conclusion, our results show that antigen-and PAF-induced pleural neutrophil and eosinophil accumulation, but not exudation, is sensitive to treatment with yangambin. In addition, yangambin also suppressed the pleural neutrophil infiltration triggered by LTB 4 as well as the blood thrombocytopenia and intestinal anaphylaxis elicited by antigen in rats. Thus, our findings indicate that yangambin shows an antagonistic action on receptors other than those of PAF, i.e., LTB 4 , and strongly suggest that it may be a useful drug in the treatment of some allergic inflammatory responses.