Short Antimicrobial Peptide Derived from the Venom Gland Transcriptome of Pamphobeteus verdolaga Increases Gentamicin Susceptibility of Multidrug-Resistant Klebsiella pneumoniae.

Infectious diseases account for nine percent of annual human deaths, and the widespread emergence of antimicrobial resistances threatens to significantly increase this number in the coming decades. The prospect of antimicrobial peptides (AMPs) derived from venomous animals presents an interesting alternative for developing novel active pharmaceutical ingredients (APIs). Small, cationic and amphiphilic peptides were predicted from the venom gland transcriptome of Pamphobeteus verdolaga using a custom database of the arthropod's AMPs. Ninety-four candidates were chemically synthesized and screened against ATCC® strains of Escherichia coli and Staphylococcus aureus. Among them, one AMP, named PvAMP66, showed broad-spectrum antimicrobial properties with selectivity towards Gram-negative bacteria. It also exhibited activity against Pseudomonas aeruginosa, as well as both an ATCC® and a clinically isolated multidrug-resistant (MDR) strain of K. pneumoniae. The scanning electron microscopy analysis revealed that PvAMP66 induced morphological changes of the MDR K. pneumoniae strain suggesting a potential "carpet model" mechanism of action. The isobologram analysis showed an additive interaction between PvAMP66 and gentamicin in inhibiting the growth of MDR K. pneumoniae, leading to a ten-fold reduction in gentamicin's effective concentration. A cytotoxicity against erythrocytes or peripheral blood mononuclear cells was observed at concentrations three to thirteen-fold higher than those exhibited against the evaluated bacterial strains. This evidence suggests that PvAMP66 can serve as a template for the development of AMPs with enhanced activity and deserves further pre-clinical studies as an API in combination therapy.

Influence of Wine pH and Ethanol Content on the Fining Efficacy of Proteins from Winemaking By-Products.

Wine color and limpidity are important aspects of consumer preferences. The alteration of these parameters can damage wine's appearance but also its mouthfeel characteristics due to its relationship with attributes such as bitterness and astringency. Fining is a practice usually used in enology to modulate undesirable wine organoleptic attributes. However, there are several factors that influence this technique. In this study, the influence of wine pH and ethanol content on grape seed protein fining efficacy has been assessed. Wine clarification, total phenolic and flavanol contents, antioxidant activity, and chromatic parameters have been investigated before and after fining process. The most noticeable clarifying effects were observed for the experimental wines with a lower pH and ethanol content. Control of these factors will make it possible to modulate the main organoleptic properties of the wine, also avoiding the addition of large amounts of fining agents and thus providing greater versatility to wineries during winemaking. Furthermore, our findings indicated that grape seed protein is a potential alternative to other plant-based fining proteins commonly used in winemaking. Its effects on clarification and color quality have been found to be comparable to those of potato protein and significantly better than those of pea protein.

Feasibility study on the use of ATR-FTIR spectroscopy as a tool for the estimation of wine polysaccharides.

Polysaccharides are the main macromolecules of colloidal nature in wines. These compounds play a critical role in stabilizing other molecules in solution and thus modifying the wine processing and organoleptic properties. Different analytical techniques have been proposed for their determination. However, most of them are complicated and time-consuming. To overcome these drawbacks, Fourier transform infrared spectroscopy (FTIR) has been evaluated in this study for the estimation of wine polysaccharides in a fast and non-destructive way. Spectral data have been correlated with wine polysaccharide contents by modified partial least squares regression (MPLS) using different spectral pretreatments. MPLS models developed have revealed the potential of FTIR analysis for the routine screening of polysaccharides rich in arabinose and galactose (PRAG), rhamnogalacturonans types II (RG-II), mannoproteins (MP) and total soluble polysaccharides (TSP) in wine samples, obtaining standard errors of prediction from 6.07 to 8.44%. Monitoring the wine polysaccharides can assist in the elaboration of the wines according to their requirements and improving quality to satisfy consumer preferences.

Assessment of Total Fat and Fatty Acids in Walnuts Using Near-Infrared Hyperspectral Imaging.

"Persian" walnut (Juglans Regia L.) is one of the most consumed tree nuts in the world. It is rich in several bioactive compounds, with polyunsaturated and monounsaturated fatty acids (PUFA and MUFA) appearing at high concentrations. Walnut consumption protects against cardiovascular, carcinogenic, and neurological disorders. The fatty acid profile has usually been determined by gas chromatography, a reliable and robust tool, but also complex, polluting, and time consuming. In this study, near infrared hyperspectral imaging has been used for the screening of total fat, MUFA, PUFA, saturated, and individual fatty acids in walnuts. Five different walnuts varieties have been considered and modified partial least square (MPLS) regressions have been performed. The SEs of prediction (SEP) in external validation (ranged from 2.12% for PUFA to 13.08% for MUFA) suggest that hyperspectral imaging can be a reliable tool for controlling these parameters in a simple, non-destructive and environmentally friendly way.

Risk of COVID-19 transmission in esophageal, anorectal manometry and 24-hour impedance-pH monitoring.

BACKGROUND: the impact of the COVID-19 pandemic has led to the interruption of most manometry or impedance-pH monitoring studies. The risk of restarting activities is unknown. OBJECTIVE: assess the risk of SARS-CoV-2 virus infection, both to patients and healthcare workers, in relation to esophageal and anorectal functional tests during the pandemic without protective measures. METHOD: a questionnaire was designed to determine whether patients and healthcare workers had COVID-19, confirmed by either a test or compatible symptoms, after functional studies were performed from January until March 2020. RESULTS: the survey was answered by 263 (92.9 %) patients. Four (1.52 %) patients had confirmed COVID-19 in the two weeks after the functional test (adjusted rate 8.34 cases per 1,000 [95 % CI -0.06-16.74], OR 0.84 [95 % CI: 0.83-0.85], p < 0.001) and no patient after anorectal manometry. Another five had only compatible symptoms, for a total of nine patients (3.42 %) (adjusted rate 27.50 cases/1,000 [95 % CI: 7.27-47.74], OR 2.84 [95 % CI: 2.81-2.87]). In the total study period, 18.25 % had confirmed COVID-19 or compatible symptoms. The average number of days between the procedure and the first day of symptoms was progressively shortened (January: 56 days, February: 33 days, March: 10.5 days). Two of ten healthcare workers (20 %) had confirmed COVID-19. CONCLUSIONS: the risk of COVID-19 infection when performing functional tests is low and more related to the evolution of the pandemic rather than to the procedure itself. The small number of healthcare workers included in the study does not allow a definitive conclusion to be drawn on their risk of infection.

Gastro-esophageal reflux disease: limits of medical treatment and surgical indications.

Proton-pump inhibitors (PPI) have long been considered as the ideal treatment for gastroesophageal reflux disease (GERD), and their limitations and side effects have revealed a need for new therapeutic approaches. At present, the therapeutic gains achieved are relatively small or limited to groups of patients with specific characteristics. This article updates the contributions, indications, and limitations of pharmacological, endoscopic, and surgical treatment.

Reduction of the Number of Samples for Cost-Effective Hyperspectral Grape Quality Predictive Models.

Developing chemometric models from near-infrared (NIR) spectra requires the use of a representative calibration set of the entire population. Therefore, generally, the calibration procedure requires a large number of resources. For that reason, there is a great interest in identifying the most spectrally representative samples within a large population set. In this study, principal component and hierarchical clustering analyses have been compared for their ability to provide different representative calibration sets. The calibration sets generated have been used to control the technological maturity of grapes and total phenolic compounds of grape skins in red and white cultivars. Finally, the accuracy and precision of the models obtained with these calibration sets resulted from the application of the selection algorithms studied have been compared with each other and with the whole set of samples using an external validation set. Most of the standard errors of prediction (SEP) in external validation obtained from the reduced data sets were not significantly different from those obtained using the whole data set. Moreover, sample subsets resulting from hierarchical clustering analysis appear to produce slightly better results.

Optimization of Protein Extraction of Oenological Interest from Grape Seed Meal Using Design of Experiments and Response Surface Methodology.

After grape processing, large amounts of seeds are produced as a side product. Grape seeds are rich in proteins that could be extracted and revalorized by the wine industry due to their high techno-functional value for modulating quality features of red wines or as an alternative to the use of other stabilizers, which are also proteins but submitted to legal restrictions. Box-Behnken design and response surface methodology were used to optimize the protein extraction from defatted grape seed meal to maximize the protein purity of the isolate under practical operating conditions of pH, temperature, meal/water ratio and extraction time. The most significant factor was pH, both in the linear and quadratic forms. Additionally, the interaction between pH and temperature exhibited a significant effect on protein content. The removal of phenolic compounds from grapeseed meal, before optimizing protein extraction, led to a significant increase of 7.70% in the protein purity of grape seed protein concentrate. Therefore, grape seeds can be used as a sustainable way for protein production in the oenological sector due to the availability of grape seeds, their low cost as a grape-processing by-product, and the protein purity reached thanks to the described procedure for optimization of protein extraction.

Response of Mycorrhizal 'Touriga Nacional' Variety Grapevines to High Temperatures Measured by Calorespirometry and Near-Infrared Spectroscopy.

Heat stress negatively affects several physiological and biochemical processes in grapevine plants. In this work, two new methods, calorespirometry, which has been used to determine temperature adaptation in plants, and near-infrared (NIR) spectroscopy, which has been used to determine several grapevine-related traits and to discriminate among varieties, were tested to evaluate grapevine response to high temperatures. 'Touriga Nacional' variety grapevines, inoculated or not with Rhizoglomus irregulare or Funneliformis mosseae, were used in this study. Calorespirometric parameters and NIR spectra, as well as other parameters commonly used to assess heat injury in plants, were measured before and after high temperature exposure. Growth rate and substrate carbon conversion efficiency, calculated from calorespirometric measurements, and stomatal conductance, were the most sensitive parameters for discriminating among high temperature responses of control and inoculated grapevines. The results revealed that, although this vine variety can adapt its physiology to temperatures up to 40 °C, inoculation with R. irregulare could additionally help to sustain its growth, especially after heat shocks. Therefore, the combination of calorespirometry together with gas exchange measurements is a promising strategy for screening grapevine heat tolerance under controlled conditions and has high potential to be implemented in initial phases of plant breeding programs.

The limited knowledge of placental damage due to neglected infections: ongoing problems in Latin America.

The placenta works as a selective barrier, protecting the fetus from potential infections that may affect the maternal organism during pregnancy. In this review, we will discuss several challenging infections that are common within Latin American countries and that may affect the maternal-fetal interface and pose risks to fetal development. Specifically, we will focus on emerging infectious diseases including the arboviruses, malaria, leishmaniasis, and the bacterial foodborne disease caused by Shiga toxin-producing Escherichia coli. We will also highlight some topics of interest currently being studied by research groups that comprise an international effort aimed at filling the knowledge gaps in this field. These topics address the relationship between exposure to microorganisms and placental abnormalities, congenital anomalies, and complications of pregnancy. ABBREVIATIONS: ADE: antibody-dependent enhancement; CCL2: monocyte chemoattractant protein-1; CCL3: macrophage inflammatory protein-1 α; CCL5: chemokine (C-C motif) ligand 5; CHIKV: chikungunya virus; DCL: diffuse cutaneous leishmaniasis; DENV: dengue virus; Gb3: glycolipid globotriaosylceramyde; HIF: hypoxia-inducible factor; HUS: hemolytic uremic syndrome; IFN: interferon; Ig: immunoglobulins; IL: interleukin; IUGR: intrauterine growth restriction; LCL: localized cutaneous leishmaniasis; LPS: lipopolysaccharid; MCL: mucocutaneous leishmaniasis; NO: nitric oxide; PCR: polymerase chain reaction; PGF: placental growth factor; PM: placental malaria; RIVATREM: Red Iberoamericana de Alteraciones Vasculares em transtornos del Embarazo; sVEGFR: soluble vascular endothelial growth factor receptor; STEC: shiga toxin-producing Escherichia coli; stx: shiga toxin protein; TNF: tumor necrosis factor; TOAS: T cell original antigenic sin; Var2CSA: variant surface antigen 2-CSA; VEGF: vascular endothelial growth factor; VL: visceral leishmaniasis; WHO: world health organization; YFV: yellow fever virus; ZIKV: Zika virus.

Valorization of American Barrel-Shoot Wastes: Effect of Post Fermentative Addition and Readdition on Phenolic Composition and Chromatic Quality of Syrah Red Wines.

The influence of post fermentative addition of American barrel-shoot wastes on phenolic composition and chromatic quality of Syrah red wines has been evaluated as an environmentally sustainable alternative to the conventional winemaking for avoiding the common color loss of red wines elaborated in warm climates. American oak wood byproducts added were previously classified by hyperspectral image analysis according to the amount of phenolic compounds transferred to the extraction media. After that, wines were elaborated under different maceration conditions by applying only one proportion of wood (12 g L-1) and two different maceration procedures (simple and double addition) and were compared with a traditionally macerated Syrah red wine (CW, no wood addition). Results proved the effectiveness of the moderate postfermentative addition of oak wood byproducts to stabilize the color of wines and to provoke lower color modification along the time, producing color wines chromatically more stable for a better aging. In the case of double addition, the adsorption of the pigments during the maceration presents a stronger effect on the color than copigmentation and polymerization by cause of the copigments extracted from the wood.

Phenolic compounds extraction in enzymatic macerations of grape skins identified as low-level extractable total anthocyanin content.

Anthocyanins in wine principally depend on grape skin extractable anthocyanin content, that is, the amount of anthocyanins present in grape skin that are released to wine during the maceration stage. This amount of extractable anthocyanins is closely linked to the cell wall degradation of skin cells. Indeed, among other methodologies, the maceration in presence of different enzymes can be used to increase cell wall degradation, and therefore, the amount of anthocyanins extracted from grape skins to wine. Vitis vinifera L. cv. Tempranillo and Syrah red grapes have been identified as samples with low anthocyanin extraction potential by near infrared hyperspectral imaging. Grape skins have been macerated in the presence of cellulase, glucosidase, and pectinase. Then, color of the supernatants and phenolic compounds extracted from grape skins (total phenols, total flavanols, and total and individual anthocyanins) has been determined. Cellulase and glucosidase have shown a positive effect in the extraction of phenolic compounds from these grapes. Macerations carried out in the presence of cellulase have produced supernatants with a more intense color (lower lightness and higher chroma values), and a higher extraction of flavanols and anthocyanins than the respective control essays. However, pectinase treatments have produced the opposite effect, which could be partially explained by an eventual interaction between the cell wall polysaccharides liberated by pectinase and the phenolic compounds extracted. Synergy effects do not appear between cellulase and glucosidase. Moreover, the negative effect of the addition of pectinase might be due to the interactions between the cell wall material liberated by pectinase and the phenolic compounds extracted. PRACTICAL APPLICATION: In the present study, grape samples with a low anthocyanin extraction potential have been identified, and these samples have been macerated in the presence of different enzymes. The applied enzymes were three of the most common enzymes that are applied in the wine industry. Individual enzymes and mixtures have been applied to Syrah and Tempranillo grape skin samples and the results have been compared to control macerations. Knowledge in this topic will help the production of quality wines.

Comparative study on the use of three different near infrared spectroscopy recording methodologies for varietal discrimination of walnuts.

Walnut fruit (Juglans regia L.) is an internationally well-known product with an important tradition of consumption. Its health benefits and economic importance in the food industry make this nut an interesting research topic. In this feasibility study, 200 walnut samples of 5 different varieties were collected and their near infrared (NIR) spectra were recorded with 3 different devices: a benchtop Fourier transform near infrared (FT-NIR) spectrograph, a dispersive hyperspectral imaging camera and a portable NIR dispersive spectrograph. Discriminant analyses were applied and different methods for the varietal discrimination of walnuts were obtained and compared. Up to 96 and 84% of correct identification were respectively obtained in internal (training set) and external validations. Better results were obtained covering the entire shell surface than collecting a unique random spectrum per sample. Moreover, FT-NIR and hyperspectral tools produced classification models with a lower classification error in internal and external validations than the portable NIR one.

Feasibility study on the use of a portable micro near infrared spectroscopy device for the "in vineyard" screening of extractable polyphenols in red grape skins.

There is substantial variation in levels of extractable phenolic compounds of red grapes (Vitis vinifera L.). Therefore, it could be desirable to known the aforesaid parameter at least for each vine. Nowadays, interest has shifted toward the development of portable vis/NIR systems, innovation in optical system design and miniaturization for its friendly use directly in the field. Spectra of intact grapes and grapes skins were recorded at harvest time in two different vintages (2016 and 2017 respectively) using a portable micro NIR spectrophotometer (908-1676 nm). A number of chemometric approaches have been used for spectral interrogation and evaluation of the aforesaid device. Spectral data have been correlated with red grape skin extractable polyphenols (total phenolic, anthocyanins and flavanols) by modified partial least squares regression (MPLS) using a number of spectral pretreatments. Moreover, different statistics strategies have been performed to develop a qualitative analysis of the data (linear discriminant analysis, discriminant partial least square analyses and Pearson's similarity index). After an exhaustive analysis of the obtained results in two different seasons, it can be concluded that the use of the portable micro NIR device for the "in vineyard" screening of extractable polyphenols in red grape skins is hampered by a number of factors. Environmental and physiological conditions should be considered to evaluate and remove factors that hamper a good sorting the berries according to their extractable polyphenol contents.

Estimation of Total Phenols, Flavanols and Extractability of Phenolic Compounds in Grape Seeds Using Vibrational Spectroscopy and Chemometric Tools.

Near infrared hyperspectral data were collected for 200 Syrah and Tempranillo grape seed samples. Next, a sample selection was carried out and the phenolic content of these samples was determined. Then, quantitative (modified partial least square regressions) and qualitative (K-means and lineal discriminant analyses) chemometric tools were applied to obtain the best models for predicting the reference parameters. Quantitative models developed for the prediction of total phenolic and flavanolic contents have been successfully developed with standard errors of prediction (SEP) in external validation similar to those previously reported. For these parameters, SEPs were respectively, 11.23 mg g-1 of grape seed, expressed as gallic acid equivalents and 4.85 mg g-1 of grape seed, expressed as catechin equivalents. The application of these models to the whole sample set (selected and non-selected samples) has allowed knowing the distributions of total phenolic and flavanolic contents in this set. Moreover, a discriminant function has been calculated and applied to know the phenolic extractability level of the samples. On average, this discrimination function has allowed a 76.92% of samples correctly classified according their extractability level. In this way, the bases for the control of grape seeds phenolic state from their near infrared spectra have been stablished.

Novel method for holocellulose analysis of non-woody biomass wastes.

Compositional analysis of non-woody biomass is quite important to determine its possible applications. However, current standard methods developed for woody biomass compositional analysis have been revealed to be unsuitable when applied to non-woody biomass. Therefore, a novel and less-time consuming modified method which enables for a proper isolation of holocellulose in non-woody biomass samples while increasing lignin degradation has been developed. The novel method mainly consists in a treatment with sodium chlorite and glacial acetic acid at boiling point, which precludes changes in the holocellulose crystallinity degree or losses of carbohydrates, as shown by DSC, XRPD, and HPLC analysis. It was successfully applied to the determination of the structural components of 10 different non-woody biomass samples. Also, its use revealed that non-woody biomass belongs to LHC and LCH groups in the biomass structural composition ternary diagram, which are completely different than the ones the woody biomass belongs to.

Evaluation of extractable polyphenols released to wine from cooperage byproduct by near infrared hyperspectral imaging.

Extractable total phenolic content of American non-toasted oak (Quercus alba L.) shavings has been determined using near infrared hyperspectral imaging. A like-wine model solution was used for the simulated maceration procedure. Calibrations were performed by partial least squares regression (MPLS) using a number of spectral pre-treatments. The coefficient of determination of wood for extractable total phenolic content was 0.89, and the standard error of prediction was 6.3 mg g-1. Thus, near infrared hyperspectral imaging arises as an attractive strategy for predicting extractable total phenolic content in the range of 0-65 mg g-1, of great relevance from the point of view of quality assurance regarding wood used in the wine sector. Near infrared hyperspectral imaging arises as an attractive strategy for the feasibility of enhancing the value of cooperage byproduct through the fast determination of extractable bioactive molecules, such as polyphenols.

Vascular Dysfunction in Mother and Offspring During Preeclampsia: Contributions from Latin-American Countries.

Pregnancy is a physiologically stressful condition that generates a series of functional adaptations by the cardiovascular system. The impact of pregnancy on this system persists from conception beyond birth. Recent evidence suggests that vascular changes associated with pregnancy complications, such as preeclampsia, affect the function of the maternal and offspring vascular systems, after delivery and into adult life. Since the vascular system contributes to systemic homeostasis, defective development or function of blood vessels predisposes both mother and infant to future risk for chronic disease. These alterations in later life range from fertility problems to alterations in the central nervous system or immune system, among others. It is important to note that rates of morbi-mortality due to pregnancy complications including preeclampsia, as well as cardiovascular diseases, have a higher incidence in Latin-American countries than in more developed countries. Nonetheless, there is a lack both in the amount and impact of research conducted in Latin America. An impact, although smaller, can be seen when research in vascular disorders related to problems during pregnancy is analyzed. Therefore, in this review, information about preeclampsia and endothelial dysfunction generated from research groups based in Latin-American countries will be highlighted. We relate the need, as present in many other countries in the world, for increased effective regional and international collaboration to generate new data specific to our region on this topic.

Study of phenolic extractability in grape seeds by means of ATR-FTIR and Raman spectroscopy.

Near infrared hyperspectral imaging has been applied to grape seeds in order to select a representative subset of samples according to their spectral features in the 900-1700nm range. Afterwards, selected grape seeds have been classified according to their total phenol and flavanol extractabilities. In this way, samples were sorted in three different groups identified as low, medium and high extractability levels. In order to establish the chemical structures which can be responsible for the different extractabilities, vibrational spectroscopy has been applied to the non-extracted material after seed extractions. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) and Raman spectra of non-extracted seed material have been recorded and their main spectral features have been linked to extractabilities of flavanolic and total phenolic compounds. The vibrational spectroscopic analysis confirms that grape seed phenolic extractability is influenced by the cell wall composition (polysaccharides, lignins, pectins) and by the degree of esterification of pectins.