The root microbiome of Salicornia ramosissima as a seedbank for plant-growth promoting halotolerant bacteria

Root−associated microbial communities play important roles in the process of adaptation of plant hosts to environment stressors, and in this perspective, the microbiome of halophytes represents a valuable model for understanding the contribution of microorganisms to plant tolerance to salt. Although considered as the most promising halophyte candidate to crop cultivation, Salicornia ramosissima is one of the least-studied species in terms of microbiome composition and the effect of sediment properties on the diversity of plant-growth promoting bacteria associated with the roots. In this work, we aimed at isolating and characterizing halotolerant bacteria associated with the rhizosphere and root tissues of S. ramosissima, envisaging their application in saline agriculture. Endophytic and rhizosphere bacteria were isolated from wild and crop cultivated plants, growing in different estuarine conditions. Isolates were identified based on 16S rRNA sequences and screened for plant-growth promotion traits. The subsets of isolates from different sampling sites were very different in terms of composition but consistent in terms of the plant-growth promoting traits represented. Bacillus was the most represented genus and expressed the wider range of extracellular enzymatic activities. Halotolerant strains of Salinicola, Pseudomonas, Oceanobacillus, Halomonas, Providencia, Bacillus, Psychrobacter and Brevibacterium also exhibited several plant-growth promotion traits (e.g., 3-indole acetic acid (IAA), 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, siderophores, phosphate solubilization). Considering the taxonomic diversity and the plant-growth promotion potential of the isolates, the collection represents a valuable resource that can be used to optimize the crop cultivation of Salicornia under different environmental conditions and for the attenuation of salt stress in non-halophytes, considering the global threat of arable soil salinization.publishe

Shedding light on the african enigma: In vitro testing of homo sapiens-helicobacter pylori coevolution

The continuous characterization of genome-wide diversity in population and case- cohort samples, allied to the development of new algorithms, are shedding light on host ancestry impact and selection events on various infectious diseases. Especially interesting are the longstanding associations between humans and certain bacteria, such as the case of Helicobacter pylori, which could have been strong drivers of adaptation leading to coevolution. Some evidence on admixed gastric cancer cohorts have been suggested as supporting Homo-Helicobacter coevolution, but reliable experimental data that control both the bacterium and the host ancestries are lacking. Here, we conducted the first in vitro coinfection assays with dual humanand bacterium-matched and -mismatched ancestries, in African and European backgrounds, to evaluate the genome wide gene expression host response to H. pylori. Our results showed that: (1) the host response to H. pylori infection was greatly shaped by the human ancestry, with variability on innate immune system and metabolism; (2) African human ancestry showed signs of coevolution with H. pylori while European ancestry appeared to be maladapted; and (3) mismatched ancestry did not seem to be an important differentiator of gene expression at the initial stages of infection as assayed here.Funds were guaranteed by the project “Advancing cancer research: from basic knowledge to application”; NORTE-01-0145-FEDER-000029; Projetos Estruturados de I & D & I, funded by Norte 2020-Programa Operacional Regional do Norte. i3S is financed by FEDER-Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020-Competitiveness and Internationalization Operational Programme (POCI), Portugal 2020, and by Portuguese funds through FCT/Ministério da Ciência, Tecnologia e Inovação in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274)

Thermal stability of the essential oils isolated from Tunisian Thymus capitatus Hoff. et Link.: Effect on the chemical composition and the antioxidant and antibacterial activities

The chemical composition, the antioxidant and the antibacterial activities of essential oils, isolated from the aerial parts of Tunisian Thymus capitatus during the flowering phase, and stored in the dark during 37 days in the oven, at 60 degrees C were evaluated. Samples taken periodically were used to evaluate the chemical composition, the antioxidant and the antibacterial activities. With some fluctuations, carvacrol (68-74%) was the major component of the oil independent of the storage period. alpha-Terpinene and gamma-terpinene decreased over time, whereas p-cymene increased in the same period. Despite the thirty-seven days of storage at 60 degrees C, T. capitatus essential oil still showed high antioxidant and stable antimicrobial activity

EVALUATION OF IN VITRO DISSOLUTION OF BENZNIDAZOLE AND BINARY MIXTURES: SOLID DISPERSIONS WITH HYDROXYPROPYLMETHYLCELLULOSE AND β-CYCLODEXTRIN INCLUSION COMPLEXES

Objective: To increase the solubility/dissolution of benznidazole (BNZ) in water using two systems: solid dispersions (SD) with hydroxypropylmethylcellulose (HPMC) and β-cyclodextrin (β-CD) inclusion complexes (IC).Methods: The samples were obtained by physical mixtures (PM), kneading (KN), evaporation (EV) and by spray-dryer (SY) atomization The analysis was based on results of in vitro dissolution and molecular modeling techniques.Results: Molecular modeling showed that BNZ can form β-CD complexes in different ways such as in an aqueous solution or a vacuum. In vitro dissolution showed significant improvement in BNZ solubility in the PM, SD and IC, and also that the β-CD IC promoted better solubility than SD with HPMC.Conclusion: Considering the data obtained, it is possible to consider the technique for the formation of β-CD IC as a more effective technique in promoting the improvement of BNZ solubility compared with getting SD with HPMC which, in turn, may increase the bioavailability of the drug and improve their pharmaceutical potential

Validation of a fluorescence in situ hybridization method using peptide nucleic acid probes for detection of Helicobacter pylori clarithromycin resistance in gastric biopsy specimens

Here, we evaluated a previously established peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) method as a new diagnostic test for Helicobacter pylori clarithromycin resistance detection in paraffin-embedded gastric biopsy specimens. Both a retrospective study and a prospective cohort study were conducted to evaluate the specificity and sensitivity of a PNA-FISH method to determine H. pylori clarithromycin resistance. In the retrospective study (n=30 patients), full agreement between PNA-FISH and PCR-sequencing was observed. Compared to the reference method (culture followed by Etest), the specificity and sensitivity of PNA-FISH were 90.9% (95% confidence interval [CI], 57.1% to 99.5%) and 84.2% (95% CI, 59.5% to 95.8%), respectively. In the prospective cohort (n=93 patients), 21 cases were positive by culture. For the patients harboring clarithromycin- resistant H. pylori, the method showed sensitivity of 80.0% (95% CI, 29.9% to 98.9%) and specificity of 93.8% (95% CI, 67.7% to 99.7%). These values likely represent underestimations, as some of the discrepant results corresponded to patients infected by more than one strain. PNA-FISH appears to be a simple, quick, and accurate method for detecting H. pylori clarithromycin resistance in paraffin-embedded biopsy specimens. It is also the only one of the methods assessed here that allows direct and specific visualization of this microorganism within the biopsy specimens, a characteristic that allowed the observation that cells of different H. pylori strains can subsist in very close proximity in the stomach. Copyright © 2013, American Society for Microbiology. All Rights Reserved.This work was supported by the Portuguese Institute Fundacao para a Ciencia e a Tecnologia (Ph.D. grant SFRH/BD/38124/2007 and project PIC/IC/82815/2007)

Allergies and Diabetes as Risk Factors for Dengue Hemorrhagic Fever: Results of a Case Control Study

Dengue is an arboviral disease that affects large areas of countries in tropical and subtropical regions of the world. Around 500,000 cases and 22,000 deaths of dengue hemorrhagic fever (DHF)/Dengue Shock Syndrome (DSS), the most severe presentations of this disease, occur annually. It is unclear why some cases of dengue fever (0.5% to 4%) progress to DHF/DSS. There is weak evidence that some diseases could have a role in this process, such as diabetes, hypertension, and allergies. In epidemics most dengue fever cases are sent home as there are too many to be kept in observation, but if it were possible to identify those with a higher risk of progression to DHF, they could be kept for observation, for early detection of signs, symptoms and alterations in laboratory tests suggestive of DHF, to enable timely and effective clinical management and early intervention. We study this issue and we believe that the evidence produced in this study, when confirmed in other studies, suggests that screening criteria might be used to identify adult patients at a greater risk of developing DHF with a recommendation that they remain under observation and monitoring in a hospital

Atherosclerosis and Bone Loss in Humans–Results From Deceased Donors and From Patients Submitted to Carotid Endarterectomy

Funding: We wish to thank all the collaborators (administrative staff, nurses, etc.) of the surgery block, as well as the doctors of the vascular surgery and transplantation departments of the Hospital of Santa Maria for the availability and assistance in the collection of the samples. We also thank Sociedade Portuguesa de Reumatologia for funding with two fellowships: Fundo de Apoio à Investigação 2014 and SPR/MSD 2015. DC-F received funding from a PhD grant from Fundação para a Ciência e a Tecnologia (SFRH/BD/80940/2011).Background and Aims: Atherosclerosis and osteoporosis share common risk factors, as well as inflammatory mechanisms. Our aim was to understand how atherosclerotic lesions are related with disturbances in bone. Methods: Gene expression of pro-inflammatory and bone metabolism related proteins (IL-1β, IL-6, IL-17A, TNF, RANKL, OPG, COL1, CTSK, OCL, TRAP, CBFA1, DKK1, SOST, ADIPOQ, and ADIPOR1) were analyzed in arteries and bones from 45 deceased donors and adipose tissue was used as control. Additionally, in 139 patients with advanced atherosclerosis submitted to carotid endarterectomy we compared calcium content (Alizarin red) and plaque inflammatory scores (CD3+, CD68+, and adiponectin) of patients with normal bone mineral density (BMD) with those with low BMD and explored the associations between gene expression in atherosclerotic plaques and BMD. Serum levels of pro-inflammatory and bone related proteins were measured both in donors and patients. Associations were investigated by the Pearson or Spearman correlation tests, and multivariate regression analyzes were performed when justified. Results: Gene expression of bone remodeling and pro-inflammatory proteins correlated positively in bone and aorta, independently of age and sex of donors, but not in adipose tissue. The expression of bone formation genes was significantly higher in atheroma plaques from endarterectomized patients with normal vs. low BMD as well as inflammatory CD68+ scores, regardless of patients' age and sex, but not of body mass index. No relationship was observed between serum levels and gene expression levels of pro-inflammatory or bone remodeling proteins. Conclusions: Our results suggest that the relationship between bones and vessels in the context of atherosclerotic disease and osteoporosis may rely on the intrinsic connection between the tissues involved, independently of disease stage. Serum measurements of pro-inflammatory and bone-remodeling proteins do not accurately translate tissue pathologic processes.publishersversionpublishe

Mineral and volatile composition of agua-mel from Portugal

Agua-mel (honey-water) is a typical honey-based product produced by the Portuguese beekeepers, particularly in southern Portugal. Agua-mel was characterized by mineral content and volatiles contents. Mineral content evaluation was performed based on a random sampling of 14 samples from a total of 16 samples provided by local producers. Mineral content showed that potassium predominated in agua-mel samples (1270-4105 mg/kg). The concentration of aluminium in one sample was tenfold higher (5.8 mg/kg) than in the remaining samples (0.3-0.6 mg/kg). Agua-mel volatiles were isolated by hydrodistillation and analysed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) from a subset of eight samples. Cluster analysis showed two poorly correlated clusters (S (corr) < 0.3). Cluster I only sample was dominated by trans-beta-ocimene (19 %), gamma-terpinene (15 %) and 2-furfural (9 %). Cluster II that included the remaining seven samples showed two moderately correlated subclusters (S (corr) < 0.5). The six samples with high correlation from subcluster IIa were dominated by 2-furfural (18-41 %) and benzene acetaldehyde (12-39 %). n-Nonadecane (14 %), n-heneicosane and 2-furfural (both 13 %) were the main components of subcluster IIb sample. Although the presence of some volatile compounds can help in the correlation between agua-mel and honey botanical source, the final product varies largely according to the preparation process even for the same producer, in different years. Agua-mel detailed characterization may assist in bringing added value to this typical Portuguese honey-based product