Effects of fungus inoculation and salt stress on physiology and biochemistry of in vitro grapevines: Emphasis on sugar composition changes by FT-IR analyses

The impacts of salt stress and inoculation in in vitro grapevine (Vitis vinifera L.) growth, nutrient accumulation, osmoregulation, photosynthesis and membrane integrity were evaluated. One month exposure to 100 mM NaCl as well as to inoculation with Phaeomoniella chlamydospora reduced relative growth rate (RGR) and induced senescence in grapevine plants, shown by: (1) decrease of Ψ_π without osmoregulation, (2) decrease of chlorophyll content and fluorescence, (3) loss of membrane integrity and (4) nutritional disorders. To assess putative changes in structural and/or non-structural carbohydrates induced by these two stress conditions, alcohol insoluble residues from the roots, stems and leaves were also characterised by FT-IR and GC with respect to the sugar composition. The referred organs were distinguished based on: (1) higher proportion of uronic acid residues in leaves which diagnose the presence of pectic polysaccharides (wavenumbers 1100, 1150 and 1018 cm^?1 in FT-IR spectra), (2) higher proportion of xylose and glucose on stems and FT-IR spectra diagnostic of xylose-rich polysaccharides (1041 cm^?1) and cellulose (1060 cm^?1), (3) higher proportion of glucose residues, xylose and arabinose on roots and a FT-IR spectra characteristic of xylose-rich polysaccharides (1041 cm^?1). The main alterations induced by salt stress and inoculation were more visible in leaves, where the content of uronic acid decreased showing that changes in cell wall composition occurred, mostly at the pectic fraction. Besides, an accumulation of insoluble glucose was found, and FT-IR spectra showed that this glucose-based material was starch (maximum absorption at 998 cm^?1), accumulated as a non-specific response to salt stress and P. chlamydospora inoculation.     

Partitioning of glycomacropeptide in aqueous two-phase systems

The partition behavior of glycomacropeptide (GMP) was determined in polyethylene glycol (PEG) and sodium citrate aqueous two-phase systems (ATPS). It was found that the partitioning of GMP depends on PEG molar mass, tie line length, pH, NaCl concentration and temperature. The obtained data indicates that GMP is preferentially partitioned into the PEG phase without addition of NaCl at pH 8.0. Larger tie line lengths and higher temperatures favor GMP partition to the PEG phase. Furthermore, it was verified that PEG molar mass and concentration have a slight effect on GMP partition. The increase in the molar mass of PEG induces a reduction of the protein solubility in the top PEG rich phase, being shown that the use of PEG1500 is beneficial for the extraction of GMP. A protein recovery higher than 85% was obtained in the top phase of these systems, clearly demonstrating its suitability as a starting point for the separation of GMP.     

Bioaccumulation of Amylose-Like Glycans by Helicobacter pylori

Background:  Helicobacter pylori cell surface is composed of lipopolysaccharides (LPSs) yielding structures homologous to mammalian Lewis O -chains blood group antigens. These structures are key mediators in the definition of host-microbial interactions and known to change their expression pattern in response to environmental pressure.
Aims:  The present work is focused on the identification of new H. pylori cell-surface glycosides. Special attention is further devoted to provide insights on the impact of in vitro subcultivation on H. pylori cell-surface phenotypes.
Methods:  Cell-surface glycans from H. pylori NCTC 11637 and two clinical isolates were recovered from the aqueous phase resulting from phenol:water extraction of intact bacteria. They were evaluated in relation to their sugars and glycosidic-linkages composition by CG-MS, size-exclusion chromatography, NMR, and Mass Spectrometry. H. pylori glycan profile was also monitored during subcultivation in vitro in agar and F12 liquid medium.
Results:  All three studied strains produce LPS expressing Lewis epitopes and express bioaccumulate amylose-like glycans. Bioaccumulation of amylose was found to be enhanced with the subcultivation of the bacterium on agar medium and accompanied by a decrease in the expression of LPS O -chains. In contrast, during exponential growth in F12 liquid medium, an opposite behavior is observed, that is, there is an increase in the overall amount of LPS and decrease in amylose content.
Conclusions:  This work shows that under specific environmental conditions, H. pylori expresses a phase-variable cell-surface α-(1→4)-glucose moiety.     

Biomphalaria spp. (Preston,1910) snails in the municipality of Juiz de Fora, Zona da Mata Mineira mesoregion, state of Minas Gerais, Brazil

This study focuses on the geographic distribution of the snail of the genus Biomphalaria and evaluates its infectivity by Schistosoma mansoni in 5264 specimens collected in the municipality of Juiz de Fora, Minas Gerais, Brazil. Of the 31 locations studied, 6 were reservoirs, 11 rudimentary holding ponds, 7 irrigation ditches, 5 lakes, 1 ornamental pond, and 1 waterfall. Intermediate hosts were found only in the rudimentary ponds and ditches, which were 100% positive. Using morphological and molecular analysis techniques, B. tenagophila, B. peregrina, and B. straminea were identified. This is the first report of B. stramínea in the municipality, and evaluation of its infective potential revealed susceptibility of 25.4%. Although we did not find specimens of Biomphalaria infected by S. mansoni, the data obtained indicate the presence of intermediate hosts, especially in the irrigation ditches in Juiz de Fora, and their proximity to contaminated areas.     

Dimeric calcium complexes of arabinan-rich pectic polysaccharides from Olea europaea L. cell walls

The study carried out in this work concerns the pectic polysaccharides of olive cell walls as present in olive pulp and that remained entrapped in the cellulosic residue after sequential extraction of the cell wall material (CWM) with imidazole, carbonate and KOH aqueous solutions. These polymers, obtained after neutralisation and dialysis of an aqueous suspension of the residue (sn-CR fraction), extracted with 4 M KOH, were arabinan-rich pectic polysaccharides. They accounted for 11–19% of the total pectic polysaccharides found in the olive pulp cell walls of fruits collected in two years and in three stages of ripening (green, cherry and black). The analysis by powder X-ray diffraction highlighted the existence, in all sn-CR fractions, of crystalline phases related with the presence of calcium-pectic polysaccharide complexes (CPPC) occurring in an amorphous carbohydrate network. The relative crystallinity of the CPPC varied linearly with the Ca²⁺/GalA molar ratio until a maximum of 0.57. Size-exclusion chromatography showed that sn-CR fractions possessed a bimodal molecular weight distribution. The lower molecular weight fraction of sn-CR (M_w = 70–135 kDa) was independent on the ripening stage of olive fruit, whereas the higher molecular weight fraction showed values of 1.1, 0.6–0.9 and 0.5–0.7 MDa, respectively, for green, cherry and black olives. Treatment of the sn-CR pectic polysaccharides with a 2 M imidazole solution disrupted the CPPC crystalline network showing the loss of low molecular weight galacturonan-rich material during dialysis (12–14 kDa cut off) and the decrease of molecular weight of the polymers to roughly half. These results allowed to infer the presence of oligogalacturonides held within cell walls by calcium ions and that the pectic polysaccharides of sn-CR fraction occurred in olive pulp cell walls as calcium bridged macrodimers.     

Casein-Derived Peptides with Antihypertensive Potential: Production,Identification and Assessment of Complex Formation with Angiotensin I-Converting Enzyme (ACE) through Molecular Docking Studies

Hypertension is nowadays one of the major world concerns in public health. Several food proteins, among which caseins, can be substrates for generating peptides with antihypertensive potential. With the increasingly evolution of computational tools, in silico molecular modeling have gained prominence in studies of protein-ligand complexes in different research fields, such as pharmaceutics and biochemical engineering. However, the application of such methodologies in food-related research can be considered still embryonic. Thus, the central aim of the present work was to apply molecular modelling in order to elucidate the molecular bases of the anti-hypertensive potential of milk caseins-derived peptides. Firstly, hydrolysates obtained from a controlled trypsinolysis of caseins were fractioned according to their molecular weight, by ultrafiltration and RP-HPLC. The obtained fractions were evaluated with regard to their in vitro inhibitory angiotensin-converting enzyme activity (%I_ACE). Six chromatographic fractions were identified, and three of them displayed high ACE-inhibition (F₁: 80.68%; F₂: 79.00%; and F₄: 62.44%). Finally, intermolecular interactions networks in complexes formed between ACE and the identified peptides were assessed through in silico molecular docking. At the molecular level, a correlation between in vitro and in silico results was found: the peptides FFVAPFPEVFGK (F₆), FALPQYLK (F₂, F₄) and ALNEINQFYQK (F₁) presented the lowest biding energies and interacted by specific H-bonds, electrostatic and hydrophobic interactions formed within ACE active site S1 residues (Ala354, Glu384, and Tyr 523) and the Zn²⁺ coordinated residues (His383, His387, and Glu411). The fraction F₃, despite its low total peptide concentration, presented a moderate inhibitory activity for ACE (49.2%), likely due to H-bonds between HQGLPQEVLNENLLR and the active site S1 residues.

     

Cognitive and Surgical Outcome in Mesial Temporal Lobe Epilepsy Associated with Hippocampal Sclerosis Plus Neurocysticercosis: A Cohort Study

Background

Where neurocysticercosis (NCC) is endemic, chronic calcified neurocysticercosis (cNCC) can be observed in patients with mesial temporal lobe epilepsy associated with hippocampal sclerosis (MTLE-HS). Considering that both disorders cause recurrent seizures or cognitive impairment, we evaluated if temporal lobectomy is cognitively safe and effective for seizure control in MTLE-HS plus cNCC.

Methods

Retrospective cohort study of neuropsychological profile and surgical outcome of 324 MTLE-HS patients submitted to temporal lobectomy, comparing the results according to the presence or absence of cNCC.

Findings

cNCC occurred in 126 (38.9%) of our MTLE-HS patients, a frequency higher than expected, more frequently in women than in men (O.R. = 1.66; 95% C.I. = 1.05–2.61; p = 0.03). Left-side (but not right side) surgery caused impairment in selected neuropsychological tests, but this impairment was not accentuated by the presence of cNCC. Ninety-four (74.6%) patients with MTLE-HS plus cNCC and 153 patients (77.3%) with MTLE-HS alone were Engel class I after surgery (O.R. = 1.16; 95% C.I. = 0.69–1.95; p = 0.58). However, the chances of Engel class IA were significantly lower in MTLE-HS plus cNCC than in patients with MTLE-HS alone (31.7% versus 48.5%; O.R. = 2.02; 95% C.I. = 1.27–3.23; p = 0.003). Patients with MTLE-HS plus cNCC showed higher rates of Engel class ID (15.1% versus 6.6%; O.R. = 2.50; 95% C.I. = 1.20–5.32; p = 0.012).

Interpretation

cNCC can be highly prevalent among MTLE-HS patients living in areas where neurocysticercosis is endemic, suggesting a cause-effect relationship between the two diseases. cNCC does not add further risk for cognitive decline after surgery in MTLE-HS patients. The rates of Engel class I outcome were very similar for the two groups; however, MTLE-HS plus cNCC patients achieved Engel IA status less frequently, and Engel ID status more frequently. Temporal lobectomy can be safely performed in most patients with MTLE-HS plus cNCC without affecting cognitive outcome. Long-term surgical seizure control in MTLE-HS plus cNCC is still satisfactory, as long as selected patients remain under medication.     

Physical Exercise Performance in Temperate and Warm Environments Is Decreased by an Impaired Arterial Baroreflex

The present study aimed to investigate whether running performance in different environments is dependent on intact arterial baroreceptor reflexes. We also assessed the exercise-induced cardiovascular and thermoregulatory responses in animals lacking arterial baroafferent signals. To accomplish these goals, male Wistar rats were subjected to sinoaortic denervation (SAD) or sham surgery (SHAM) and had a catheter implanted into the ascending aorta to record arterial pressure and a telemetry sensor implanted in the abdominal cavity to record core temperature. After recovering from these surgeries, the animals were subjected to constant- or incremental-speed exercises performed until the voluntary interruption of effort under temperate (25° C) and warm (35° C) conditions. During the constant-speed exercises, the running time until the rats were fatigued was shorter in SAD rats in both environments. Although the core temperature was not significantly different between the groups, tail skin temperature was higher in SAD rats under temperate conditions. The denervated rats also displayed exaggerated increases in blood pressure and double product compared with the SHAM rats; in particular, in the warm environment, these exaggerated cardiovascular responses in the SAD rats persisted until they were fatigued. These SAD-mediated changes occurred in parallel with increased variability in the very low and low components of the systolic arterial pressure power spectrum. The running performance was also affected by SAD during the incremental-speed exercises, with the maximal speed attained being decreased by approximately 20% in both environments. Furthermore, at the maximal power output tolerated during the incremental exercises, the mean arterial pressure, heart rate and double product were exaggerated in the SAD relative to SHAM rats. In conclusion, the chronic absence of the arterial baroafferents accelerates exercise fatigue in temperate and warm environments. Our findings also suggest that an augmented cardiovascular strain accounted for the early interruption of exercise in the SAD rats.