Serine protease inhibition and modulatory-antibiotic activity of the proteic extract and fractions from Amburana cearensis.

This study investigated the inhibitory activity of serine protease, as well as antibacterial and antibiotic modifying activities of the crude extract and fractions of A. cearensis seeds. Microdilution assay was used to evaluate the antibacterial activity and the antibiotic resistance-modulating effects of samples against multiresistant bacteria Staphylococcus aureus (SA10) and Escherichia coli (EC06). In the inhibition test for serine protease, all the samples showed inhibition of enzymatic activity. Crude extract and fractions of A. cearensis seeds showed a Minimum Inhibitory Concentration ≥1024 µg/mL for all microorganisms tested. However, the samples acted as resistance modifying agent, presenting synergism when associated with gentamicin, norfloxacin and penicillin. The present study provides data indicating a possible use of the seeds extract of A. cearensis in association with antibiotics in the fight against bacterial infections.

Polymorphism in Mitochondrial Group I Introns among Cryptococcus neoformans and Cryptococcus gattii Genotypes and Its Association with Drug Susceptibility.

Cryptococcosis, one of the most important systemic mycosis in the world, is caused by different genotypes of Cryptococcus neoformans and Cryptococcus gattii, which differ in their ecology, epidemiology, and antifungal susceptibility. Therefore, the search for new molecular markers for genotyping, pathogenicity and drug susceptibility is necessary. Group I introns fulfill the requisites for such task because (i) they are polymorphic sequences; (ii) their self-splicing is inhibited by some drugs; and (iii) their correct splicing under parasitic conditions is indispensable for pathogen survival. Here, we investigated the presence of group I introns in the mitochondrial LSU rRNA gene in 77 Cryptococcus isolates and its possible relation to drug susceptibility. Sequencing revealed two new introns in the LSU rRNA gene. All the introns showed high sequence similarity to other mitochondrial introns from distinct fungi, supporting the hypothesis of an ancient non-allelic invasion. Intron presence was statistically associated with those genotypes reported to be less pathogenic (p < 0.001). Further virulence assays are needed to confirm this finding. In addition, in vitro antifungal tests indicated that the presence of LSU rRNA introns may influence the minimum inhibitory concentration (MIC) of amphotericin B and 5-fluorocytosine. These findings point to group I introns in the mitochondrial genome of Cryptococcus as potential molecular markers for antifungal resistance, as well as therapeutic targets.

α-bisabolol-loaded lipid-core nanocapsules reduce lipopolysaccharide-induced pulmonary inflammation in mice.

Acute respiratory distress syndrome (ARDS) is a severe clinical condition of respiratory failure due to an intense inflammatory response with different etiologies. Despite all efforts, therapy remains limited, and ARDS is still associated with high mortality and morbidity. Plants can provide a vast source of active natural products for the discovery of new drugs. α-bisabolol (α-bis), a constituent of the essential oil from chamomile, has elicited pharmacological interest. However, the molecule has some limitations to its biological application. This study was conducted to develop a drug delivery system using lipid-core nanocapsules (LNCs) to improve the anti-inflammatory effects of orally administered α-bis. α-bis-loaded LNCs (α-bis-LNCs) were prepared by interfacial deposition of poly(ε-caprolactone) and orally administered in a mouse model of ARDS triggered by an intranasal administration of lipopolysaccharide (LPS). We found that α-bis-LNCs (30, 50, and 100 mg kg-1) significantly reduced airway hyperreactivity (AHR), neutrophil infiltration, myeloperoxidase activity, chemokine levels (KC and MIP-2), and tissue lung injury 18 hours after the LPS challenge. By contrast, free α-bis failed to modify AHR and neutrophil accumulation in the bronchoalveolar lavage effluent and lung parenchyma and inhibited elevation in the myeloperoxidase and MIP-2 levels only at the highest dose. Furthermore, only α-bis-LNCs reduced LPS-induced changes in mitogen-activated protein kinase signaling, as observed by a significant reduction in phosphorylation levels of ERK1/2, JNK, and p38 proteins. Taken together, our results clearly show that by using LNCs, α-bis was able to decrease LPS-induced inflammation. These findings may be explained by the robust increase of α-bis concentration in the lung tissue that was achieved by the LNCs. Altogether, these results indicate that α-bis-LNCs should further be investigated as a potential alternative for the treatment of ARDS.

Would Nesotriatoma bruneri Usinger, 1944 be a valid species?

Variation in the size and morphological differences intraspecific of Nesotriatoma flavida led to the description of N. bruneri. However, two years later the same author proposed the synonymization N. bruneri with N. flavida. N. bruneri was revalidated through morphological analysis after 35 years. Thus, given the existing taxonomic questioning between these Cuban triatomines, we analyzed new parameters such as genetic distance from the mitochondrial 16S rDNA deposited in Genbank and cytogenetic characterization, through the constitutive heterochromatin pattern, in order to reassess the specific status of N. bruneri. The analysis of the disposition of constitutive heterochromatin in the genome of these triatomines allowed observing that only the sex chromosome Y is heterochromatic, and the autosomes and the sex chromosomes X are euchromatic. These characteristics are identical to those described for N. flavida. By means of analysis of genetic distance matrix, we found that the genetic distance between N. bruneri and N. flavida was only 0.04%. Thus, by means of extremely low genetic distance and identical cytogenetic characteristics, we suggest that N. bruneri should be back again synonymized with N. flavida. However, we recommend that experimental hybrid crosses and new molecular analysis should be conducted, focusing mainly in the genetic distance based on other genes, on the rate of fertility of eggs and viability of hybrids to confirm the proposed of synonymization.

New arrangements on several species subcomplexes of Triatoma genus based on the chromosomal position of ribosomal genes (Hemiptera - Triatominae).

The hemipteran subfamily Triatominae includes 150 blood-sucking species, vectors of Chagas disease. By far the most specious genus is Triatoma, assembled in groups, complexes and subcomplexes based on morphological similarities, geographic distribution and genetic data. However, many molecular studies questioned the species integration of several subcomplexes as monophyletic units. In triatomines, chromosomal position of major ribosomal DNA (rDNA) loci is extremely variable but seems to be species-specific and an evolutionary conserved genetic trait, so that closely related species tend to have ribosomal clusters in the same chromosomal location. Considering that the autosomal position as the ancestral character for all heteropteran species, including triatomines, we suggest that the movement of rDNA loci from autosomes to sex chromosomes rapidly established reproductive barriers between divergent lineages. We proposed that the rDNA translocation from the autosomes to the sex chromosomes restrict reproductive compatibility and eventually promote speciation processes. We analyzed the chromosomal position of 45S rDNA clusters in almost all species of the matogrossensis, rubrovaria, maculata and sordida subcomplexes. The fluorescent in situ hybridization results are discussed considering the available genetic data and we proposed new arrangements in the species that constitute each one of these subcomplexes.

Modulation of the antibiotic activity against multidrug resistant strains of 4-(phenylsulfonyl) morpholine.

The compound 4-(Phenylsulfonyl) morpholine belongs to the class of sulfonamides, which are widely used in the treatment of a large number of diseases caused by microorganisms. This compound has a morpholine group, which is also known for its antimicrobial properties. The aim of the present study was to investigate the antimicrobial and modulating activity of 4-(Phenylsulfonyl) morpholine against standard and multi-resistant strains of Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and strains of the fungi Candida albicans, C. tropicalis and C. krusei. Antimicrobial activity was assessed based on the minimum inhibitory concentration (MIC) using the microdilution method. MIC was ⩾1024 µg/mL for all microorganisms. Regarding modulating activity, the most representative effect occurred with the combination of 4-(Phenylsulfonyl) morpholine at a concentration of 128 µg/mL (MIC 1/8) and amikacin against P. aeruginosa 03, with a reduction in MIC from 312.5 to 39.06 µg/mL.

Avaliação comparativa da modulação de antibióticos, frente às cepas bacterianas de Escherichia coli, Staphylococcus aureus / Evaluación comparativa de la modulación de antibióticos frente a cepas bacterianas de Escherichia Coli, Staphylococcus / Comparative Evaluation of the Modulation of Antibiotic-Activity against Strains of Escherichia coli and Staphylococcus aureus

Introdução: Combinações múltiplas de drogas estão sendo utilizadas no combate à disseminação de bactérias patogênicas resistentes a antibióticos. No entanto, a utilização incorreta desses antibióticos fez com que algumas bactérias sejam selecionadas, deixando apenas as resistentes aos agentes antimicrobianos, com o consequente surgimento de resistência aos antibióticos. Objetivos: Nesse contexto, avaliaram-se as atividades microbiológicas dos antibióticos claritromicina, gentamicina, ciprofloxacino e imipenem, além de comparar as atividades da associação entre esses fármacos, frente às cepas bacterianas de Escherichia coli e Staphylococcus aureus. Materiais e Métodos: A Concentração Inibitória Mínima (CIM) foi realizada pelo método de microdiluição em caldo para cada uma das amostras selecionadas. Posteriormente, com base nos valores das cims, foi avaliado o efeito antimicrobiano da combinação das drogas pelo método de modulação. Resultados: A associação demonstrou efeitos sinérgicos frente às linhagens testadas, sugerindo-se que a associação dos antibióticos testados aumenta o seu potencial antimicrobiano. Conclusão: Nesse sentido, o uso de antibióticos em combinação pode ser realizado mediante investigações posteriores que comprovem a segurança de seu uso.

Introducción: Actualmente, se está utilizando la combinación de múltiples drogas en la lucha contra la propagación de bacterias patógenas resistentes a los antibióticos. Sin embargo, el uso inadecuado de estos antibióticos ha causado que algunas bacterias se seleccionen, con la consiguiente aparición de resistencia a los antibióticos. Objetivo: Se evaluó la actividad microbiológica de los antibióticos claritromicina, gentamicina, ciprofloxacino e imipenem, y se comparó la actividad sinérgica de estos fármacos, frente a las cepas bacterianas de Escherichia coli y Staphylococcus aureus. Materiales y Métodos: Se determinó la Concentración Mínima Inhibitoria (CMI) por el método de microdilución en caldo para cada una de las muestras seleccionadas y se evaluó el efecto antimicrobiano de la combinación de fármacos por el método de modulación. Resultados: La combinación mostró efectos sinérgicos frente a las cepas ensayadas, lo que sugiere que la asociación de los antibióticos ensayados aumenta su potencial antimicrobiano. Conclusión: Se puede utilizar antibióticos en combinación; sin embargo, se requiere realizar investigaciones adicionales que demuestren ser seguros para su utilización.

Introduction: Combining multiple drugs is an strategy used to combat the dissemination of pathogenic and drug resistant bacteria. However, the misuse of these drugs against bacteria have caused the selection of more resistant specimens called multidrug-resistant bacteria. Objective: In this work we evaluated the antibiotic activity of claritromicin, gentamicin, ciprofloxacin and imipenen, alone or associating one by one, against strains of Escherichia coli and Staphylococcus aureus. Material and methods: The minimal inhibitory concentration (mic) was performed using the microdilution assay. Based in the mic values, the antibiotic effect of the drugs alone and in association were determined. Results: The association between the drugs demonstrated the synergism against the bacterial strains. Conclusion: The use of the combined antibiotic-therapy can be positively performed, but additional studies have to be conducted first for proving that its use is safe.

Identification and modulatory activity assessment of 2-hydroxy-3,4,6-trimethoxyacetophenone isolated from croton anisodontus müll. Arg.(euphorbiaceae).

The n-hexane extract of the stem bark of Croton anisodontus yielded 2-hydroxy-3,4,6-trimethoxyacetophenone, a well-known substance, but isolated from this species for the first time. The antimicrobial and modulatory activities of the compound towards Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, C. krusei and C. tropicalis strains were assessed. Antibiotics such as amikacin, gentamicin and neomycin were used in a sub-inhibitory concentration. Significant activity was observed towards P. aeruginosa and S. aureus 358, with p < 0.001 in association with amikacin. The present results place C anisodontus as an alternative source of 2-hydroxy-3,4,6-trimethoxyacetophenone with antibacterial potential.

In vivo measurement of the pressure signal in the intervertebral disc of an anesthetized sheep.

The purpose of the present study was to measure the intradiscal pressure signal of an anesthetized sheep under spontaneous breathing. An ultra-miniature fiber optic high-pressure sensor was implanted into the nucleus pulposus of the fifth lumbar intervertebral using a dorsolateral transforaminal approach. Results suggested the periodicity of the intradiscal pressure signal was similar to the mean respiratory rate of the animal. The average resting intradiscal pressure was also calculated and compared to available data.