[Critical patients COVID-19 has changed the management and outcomes in the ICU after 1 year of the pandemic? A multicenter, prospective, observational study]. / Pacientes críticos COVID-19. ¿Han variado el manejo y los resultados en la UCI tras un año de pandemia? Estudio multicéntrico, prospectivo, observacional.

Objective: To compare the clinical characteristics, treatments, and evolution of critical patients with COVID-19 pneumonia treated in intensive care units (ICU) after one year of pandemic. Methodology: Multicenter, prospective study, which included critical COVID-19 patients in 9 ICUs in northwestern Spain. The clinical characteristics, treatments, and evolution of patients admitted to the ICU during the months of March-April 2020 (period 1) were compared with patients admitted in January-February 2021 (period 2). Results: 337 patients were included (98 in period 1 and 239 in period 2). In period 2, fewer patients required invasive mechanical ventilation (IMV) (65% vs. 84%, P < .001), using high-flow nasal cannulas (CNAF) more frequently (70% vs. 7%, P < .001), ventilation non-invasive mechanical (NIMV) (40% vs. 14%, P < .001), corticosteroids (100% vs. 96%, P = .007) and prone position in both awake (42% vs. 28%, P = .012), and intubated patients (67% vs. 54%, P = .034). The days of IMV, ICU stay and hospital stay were lower in period 2. Mortality was similar in the two periods studied (16% vs. 17%). Conclusions: After one year of pandemic, we observed that in patients admitted to the ICU, CNAF, NIMV, use of the prone position, and corticosteroids have been used more frequently, reducing the number of patients in IMV, and the length of stay in the ICU and hospital stay. Mortality was similar in the two study periods.

Can Satellite Remote Sensing Assist in the Characterization of Yeasts Related to Biogeographical Origin?

Biogeography is a key concept associated with microbial terroir, which is responsible for the differentiation and uniqueness of wines. One of the factors influencing this microbial terroir is the vegetation, which in turn is influenced by climate, soil, and cultural practices. Remote sensing instruments can provide useful information about vegetation. This study analyses the relationship between NDVI, calculated using Sentinel-2 and Landsat-8 satellite images of different veraison dates, and microbial data obtained in 2015 from 14 commercial (organic and conventional) vineyards belonging to four Designations of Origin (DOs) from Galicia (northwest Spain). Microbial populations in grapes and musts were identified using PCR techniques and confirmed by sequencing. Statistical analyses were made using PCA, CCA, TB-PLS, and correlation analyses. This study confirms that the NDVI is positively correlated with the diversity of yeasts, both in grapes' surface and must samples. Moreover, the results of this study show: (i) Sentinel-2 images, as well as Landsat-8 images, can establish differences in NDVI related to yeast terroir in grapes and musts, as it is the most relevant DO factor, (ii) Sentinel-2 NDVI and yeast biogeography are moderately to strongly correlated, (iii) Sentinel-2 achieved a better delimitation of the DOs than Landsat-8 and can establish more accurate differences in NDVI-yeast terroir correlations, and (iv) a higher NDVI was associated with the yeast biogeographical patterns of the DOs with higher species richness (S) consisting of weakly fermenting yeasts (Hanseniaspora uvarum, Pichia spp., Starmerella bacillaris, and Zygosaccharomyces spp). However, NDVI values did not correlate well with biogeographic patterns of yeasts previously studied at frequency level (proportion or percentage of each species) in each particular DO. This study suggests that satellite imagery has the potential to be a valuable tool for wine quality management and a decision-making instrument for DO regulators and winegrowers.

Effectiveness of a group intervention using pain neuroscience education and exercise in women with fibromyalgia: a pragmatic controlled study in primary care.

BACKGROUND: Very positive effects have been described in the application of pain neuroscience education (PNE) to chronic pain and migraine. However, there are few data on the applicability of this therapeutic approach in actual clinical practice in a primary care (PC) setting. The aim of this study was to explore the efficacy in fibromyalgia (FM) of an intervention based on PNE and exercise compared to treatment as usual (TAU). METHODS: Pragmatic nonrandomised controlled trial set in 5 healthcare centres and one physiotherapy centre in PC. Fifty-three women with FM (2010 American College of Rheumatology Diagnostic Criteria for Fibromyalgia) were studied, 35 in the intervention group (IG) and 18 in the control group (CG). The women in the IG were interviewed individually and then received 6 weekly sessions plus one review session (1 month later): those in the CG received their TAU. The subject assignation to the CG or the IG was determined according to their availability to attend the sessions. They all filled in several questionnaires (prior to and 1 year after the intervention) to evaluate the impact of FM in their daily lives, catastrophism, anxiety and depression, severity and impact of pain in daily personal performance and functional capacity. RESULTS: The reductions (improvements) in the scores of all tests (baseline-final) were greater in the IG (p < 0.05) when adjusted for age and baseline values, with moderate or high effect size. After 1 year, 20% (CI - 1 to 42%) more women in the IG, compared to the CG, had a FIQ score < 39 (mild functional impairment). 17/38 (49%) women in the IG no longer met FM criteria at the end of follow-up. CONCLUSIONS: An intervention based on PNE and exercise in patients with FM is feasible and seems effective in PC. TRIAL REGISTRATION: The study was retrospectively registered at ClinicalTrials.gov (Trial Registration NCT04539171 ), on 04/09/2020.

Oenological potential of non-Saccharomyces yeasts to mitigate effects of climate change in winemaking: impact on aroma and sensory profiles of Treixadura wines.

The effects of climate change on wine include high-alcohol content, low acidity and aroma imbalance. The potential of several non-Saccharomyces wine yeasts to mitigate these effects was evaluated by sequential fermentation of Treixadura grape must. Fermentations with only Saccharomyces cerevisiae ScXG3 and a spontaneous process were used as control assays. All yeast strains were obtained from the yeast collection of Estación de Viticultura e Enoloxía de Galicia (EVEGA), Galicia, Spain. Fermentation kinetics as well as yeast dynamics and implantation ability varied depending on inoculated yeasts. In addition, the results showed significant differences in the chemical composition of wine. Starmerella bacillaris 474 reduced the alcohol content (1.1% vol) and increased the total acidity (1.2 g L-1) and glycerol of wines. Fermentation with Lachancea thermotolerans Lt93 and Torulaspora delbrueckii Td315 also decreased the alcohol content, although to a lesser extent (0.3% and 0.7% vol, respectively); however, their effect on wine acidity was less significant. The wines also differed in their concentration of volatile compounds and sensory characteristics. Thus, wines made with Metschnikowia fructicola Mf278 and S. cerevisiae ScXG3 had higher content of esters, acetates and some acids than other wines, and were most appreciated by tasters due to their fruity character and overall impression.

Effect of mixed species alcoholic fermentation on growth and malolactic activity of lactic acid bacteria.

In recent years, there is an increasing interest from the winemaking industry for the use of mixed fermentations with Starmerella bacillaris (synonym Candida zemplinina) and Saccharomyces cerevisiae, due to their ability to modulate metabolites of oenological interest. The current study was carried out to elucidate the effect of this fermentation protocol on the growth and malolactic activity of lactic acid bacteria (LAB) used for malolactic fermentation (MLF) and on the chemical and volatile profile of Nebbiolo wines and their chromatic characteristics. To this end, two LAB species, namely Lactobacillus plantarum and Oenococcus oeni, were inoculated at the beginning and at the end of the alcoholic fermentation (AF) performed by pure and mixed yeast using the abovementioned yeasts. The different yeast inoculation protocols and the combination of species tested influenced greatly the interactions and behavior of the inoculated yeasts and LAB during AF and MLF. For both LAB species, inoculation timing was critical to how rapidly MLF started and finished. Fermentation inoculated with L. plantarum, at the beginning of the AF, completed MLF faster than those inoculated with O. oeni. The presence of Starm. bacillaris in mixed fermentation promoted LAB growth and activity, in particular, O. oeni. Furthermore, LAB species choice had a greater impact on the volatile and chromatic profile of the wines than inoculation time. These findings reveal new knowledge about the importance of LAB species choice and inoculation time to ensure fast MLF completion and to improve wine characteristics in mixed fermentation with Starm. bacillaris and S. cerevisiae.

Preliminary Study on Yeasts Associated with the Production of "Tostado"-a Traditional Sweet Wine from Galicia (NW Spain).

BACKGROUND: Tostado is a traditional sweet wine from the Designations of Origins (DOs) of Ribeiro and Valdeorras in Galicia (NW Spain). The harvested grapes are air-dried and pressed to increase the concentrations of sugars, acids, and flavour compounds. Therefore, knowledge of the yeasts involved in fermentation under these conditions is essential to guarantee the quality and uniqueness of the valuable, distinctive, and expensive Tostado wines. METHODS: Saccharomyces and non-Saccharomyces yeasts were identified using Wallerstein Laboratory (WL) Nutrient Agar and lysine plating, followed by polymerase chain reaction (PCR) amplification, enzymatic digestion, and sequencing. Saccharomyces cerevisiae isolates were further characterised at the strain level using mitochondrial DNA (mtDNA) restriction fragment length polymorphism (RFLP). Statistical analyses were also performed, including different diversity indices, Similarity Percentage (SIMPER) analysis, principal component analysis (PCA), neighbor-joining clustering, parsimony-phylogram, and network plot. In addition, the total acidity, volatile acidity, reducing sugars, and alcoholic strength by volume of the Tostado wines were analysed. RESULTS: A wide diversity of autochthonous yeasts was found, which were predominantly species of oenological relevance, such as Lachancea thermotolerans, Starmerella bacillaris, Hanseniaspora uvarum, Debaryomyces hansenii, Torulaspora delbrueckii, Pichia spp., and Saccharomyces cerevisiae from the must and paste stages of Tostado wine. In addition, 19 different S. cerevisiae strains were identified. This high yeast diversity, which changed from the early stages of fermentation, could contribute to the distinctive characteristics observed in Tostado wine. CONCLUSIONS: Characteristic and differentiating chemical and microbiological profiles were found as early as the pre-fermentation stages, which adds value to these special wines that have rarely been studied.

Physicochemical Properties and Microbiome of Vineyard Soils from DOP Ribeiro (NW Spain) Are Influenced by Agricultural Management.

Agricultural management influences the soil ecosystem by affecting its physicochemical properties, residues of pesticides and microbiome. As vineyards grow crops with the highest incidence of pesticides, the aim of this study was to evaluate the impact of conventional and sustainable management systems of vineyards from DOP Ribeiro on the soil's condition. Samples from soils under three different management systems were collected, and the main soil physicochemical properties were evaluated. A selection of 50 pesticides were investigated by liquid chromatography with tandem mass spectrometry. The bacterial and fungal microbiomes were characterized through amplicon sequencing. The results show that organic agriculture positively influences soil pH and the concentration of some nutrients compared to conventional management. Our microbiome analysis demonstrated that transitioning from conventional to organic management significantly improves several BeCrop® indexes related to key microbial metabolism and soil bio-sustainability. Such a transition does not affect soil alpha diversity, but leads to a higher interconnected microbial network structure. Moreover, differential core genera and species for each management system are observed. In addition, the correlation of the microbiome with geographical distance is evidence of the existence of different microbial terroirs within DOP Ribeiro. Indeed, sustainable management leads to higher nutrient availability and enhances soil health in the short term, while lowering pesticide usage.

Fermentative aroma compounds and sensory profiles of Godello and Albariño wines as influenced by Saccharomyces cerevisiae yeast strains.

BACKGROUND: Yeasts responsible for fermentation have an important repercussion on wine quality. This study presents the influence of two autochthonous strains of Saccharomyces cerevisiae (XG1 and XG3), a commercial yeast (QA23) and spontaneous fermentation on the chemical and sensory properties of wines from Godello and Albariño. RESULTS: All the yeasts showed normal fermentative kinetics and were able to lead fermentations; therefore, they were responsible for wine chemical and sensory characteristics. Significant differences were found at the chemical level depending on yeast strain and variety. Albariño wines from XG1 and XG3 presented low total acidity and glycerol content. Godello wines from QA23 had higher total acidity but lower alcohol content than those from XG1, XG3 and spontaneous fermentation. QA23 wines presented a greater amount of higher alcohols and 2-phenylethanol for both grapevine cultivars, whereas XG3 and spontaneous fermentations yielded wines with a higher concentration of esters, mainly ethyl lactate, and fatty acids. These differences were detected at the sensory level; thus, Albariño made with XG3 and Godello from spontaneous fermentation were the most appreciated wines. CONCLUSION: XG3, an autochthonous strain of S. cerevisiae, constitutes a useful tool to elaborate wines with singular characteristics.

Inoculation of Treixadura musts with autochthonous Saccharomyces cerevisiae strains: fermentative performance and influence on the wine characteristics.

The fermentative ability of five autochthonous Saccharomyces cerevisiae strains (XG1, XG2, XG3, XG4 and XG5) and their influence on the chemical composition and sensory properties of Treixadura wines were evaluated. The inoculated strains have successfully led and completed the fermentations. Wines obtained from different yeasts showed significant differences in total and volatile acidity. Regarding volatile compounds, significant differences among wines were found for acetates, ethyl esters, acetoin, 1-hexanol, and fatty acids. Wines from spontaneous fermentation and those made with yeasts XG3 and XG4 were clearly separated through principal component analysis. Chemical composition influenced sensory properties of wines, especially at the olfactory level. Different connotations of fruity notes predominated depending on the strain. The wine from strain XG4 was the most appreciated by panelists. Our results confirmed that autochthonous S. cerevisiae strains are useful tools in winemaking because they allow obtaining singular wines from a given variety.

Characterization of Indigenous Non-Saccharomyces Yeast Strains with Potential Use in Winemaking.

BACKGROUND: The vineyard is a great reservoir of autochthonous yeast strains whose composition is defined by different regional (edaphology, orography or climatology) and anthropological factors (cultivation systems or cultural practices). Most of this yeast diversity corresponds to non-Saccharomyces strains, some of which have potential use in winemaking. METHODS: The oenological potential of 29 different native non-Saccharomyces strains belonging to 4 species (Lachancea thermotolerans, Torulaspora delbrueckii, Starmerella bacillaris and Metschnikowia spp.) was evaluated, using the autochthonous Saccharomyces cerevisiae XG3 strain as a control. Microfermentations with pure culture of each strain were performed in duplicate and the basic parameters and major volatiles of wines were analysed following official methodology. The best strain within each species was selected using a quantification matrix including the relevant oenological characteristics. RESULTS: The fermentative ability of non-Saccharomyces was lower than S. cerevisiae in all cases, but with differences among species. L. thermotolerans and T. delbrueckii showed higher fermentation rates than Starm. bacillaris, whereas Metschnikowia spp. presented a low fermentative power. At chemical level all non-Saccharomyces strains reduced the alcoholic content, the higher alcohols and the volatile acidity of wines and increased the content of glycerol, with differences among strains within a given species. T. delbrueckii and L. thermotolerans increased the total acidity of wines. The latter and Metschnikowia spp. strains produced lactic acid, which decreased the wine pH in the case of L. thermotolerans. According to their oenological traits the best rated strains of each species were Lt93, Td315, Mf278 and Sb474. In addition, the data obtained in pure fermentations were correlated to those chemical and aromatic compounds obtained with these non-Saccharomyces strains in sequential fermentations. CONCLUSIONS: Autochthonous strains of non-Saccharomyces yeast species contribute distinctive chemical characteristics to the wines. The correlations observed between wines fermented with the different non-Saccharomyces indigenous strains in pure and sequential fermentations suggest that their contribution to wine properties remains stable regardless of must composition or winemaking techniques.

Histological artifacts associated with laser and electroscalpel gingivectomy: Case series / Artefactos histológicos asociados a gingivectomía con láser y electrobisturí: serie de casos.

Introduction. Over time, efforts have been invested in the design of new instruments that overcome the disadvantages of the gold standard instrument in surgery, the scalpel. As a result, electronic equipment has emerged such as the electric scalpel and laser devices. The available evidence on these instruments suggests that the tissue response is related to each instrument's physical and biological cutting principles. Objective. To compare the histological changes in gingiva samples associated with surgical cutting performed with a 940 nm diode laser, a 2780 nm erbium, chromium: yttriumscandium-gallium-garnet (Er,Cr:YSGG) laser, and an electric scalpel, by presenting a series of cases. Case presentation. We present three cases of healthy patients undergoing cosmetic surgery. The clinical examination revealed exposure of a keratinized gingiva band greater than 4 mm, normal color and texture in gingival tissue, with a firm consistency and no bleeding on periodontal probing. Gingivectomy was indicated with the following protocols: Diode laser of 940 nm at 1 W, in continuous mode; Er,Cr:YSGG laser of 2780 nm at 2.5 W, 75 Hz, H mode, air 20, water 40, gold tip MT4); and electric scalpel in cutting mode at power level four. Gingival tissue samples were taken and stored in 10% formaldehyde for histological analysis. Conclusion. All the evaluated cutting instruments generated histological changes produced by the thermal effect, the main ones being collagen coagulation and carbonization. The depth of thermal damage caused by the 2780 nm Er,Cr:YSGG laser was much lesser than that induced by the electric scalpel and the 940 nm diode laser.

Introducción. Históricamente se ha invertido esfuerzo en el diseño de nuevos instrumentos que superen las desventajas del estándar de referencia en cirugía, el bisturí. Como consecuencia de esto, han surgido equipos electrónicos como el electrobisturí y los diferentes dispositivos de tecnología láser. La información disponible sobre estos instrumentos sugiere que la respuesta del tejido intervenido está influenciada por los principios físicos y biológicos de corte del instrumento. Objetivo. Comparar los cambios histológicos en muestras de encía asociados al corte quirúrgico realizado con láser de diodo de 940 nm, láser de erbio, cromo: itrio-escandiogalio-granate (Er,Cr:YSGG) (2780nm) y electrobisturí mediante una presentación de serie de casos. Presentación de los casos. Se presentan tres casos de pacientes sanos sometidos a cirugía estética. El examen clínico reveló la exposición de una banda gingival queratinizada mayor de 4 mm, tejido gingival de color y textura normales, de consistencia firme y sin sangrado al sondaje periodontal. Se indicó gingivectomía con los siguientes protocolos: láser de diodo de 940 nm a 1 W, en modo continuo; láser de Er,Cr:YSGG de 2780 nm a 2,5 W, 75 Hz, modo H, aire 20, agua 40, punta de oro MT4; y bisturí eléctrico en modo de corte, a nivel de potencia cuatro. Se tomaron muestras de tejido gingival y se almacenaron en formaldehído al 10 % para su análisis histológico. Conclusión. Los tres instrumentos de corte generaron cambios histológicos producidos por el efecto térmico; los principales fueron coagulación del colágeno y carbonización.

Critical patients COVID-19 has changed the management and outcomes in the ICU after 1 year of the pandemic? A multicenter, prospective, observational study.

OBJECTIVE: To compare the clinical characteristics, treatments, and evolution of critical patients with COVID-19 pneumonia treated in Intensive Care Units (ICU) after one year of pandemic. METHODOLOGY: Multicenter, prospective study, which included critical COVID-19 patients in 9 ICUs in northwestern Spain. The clinical characteristics, treatments, and evolution of patients admitted to the ICU during the months of March-April 2020 (period 1) were compared with patients admitted in January-February 2021 (period 2). RESULTS: 337 patients were included (98 in period 1 and 239 in period 2). In period 2, fewer patients required invasive mechanical ventilation (IMV) (65% vs 84%, p < 0.001), using high-flow nasal cannulas (CNAF) more frequently (70% vs 7%, p < 0.001), ventilation non-invasive mechanical (NIMV) (40% vs 14%, p < 0.001), corticosteroids (100% vs 96%, p = 0.007) and prone position in both awake (42% vs 28%, p = 0.012), and intubated patients (67% vs 54%, p = 0.034). The days of IMV, ICU stay and hospital stay were lower in period 2. Mortality was similar in the two periods studied (16% vs 17%). CONCLUSIONS: After 1 year of pandemic, we observed that in patients admitted to the ICU, CNAF, NIMV, use of the prone position, and corticosteroids have been used more frequently, reducing the number of patients in IMV, and the length of stay in the ICU and hospital stay. Mortality was similar in the two study periods.

Impact of a pharmaceutical intervention on influenza vaccination in patients receiving treatment with biological medicines.

OBJECTIVE: The aims of this study were twofold - to determine the impact of a health education intervention led by a hospital pharmacy department on influenza vaccination take-up in patients diagnosed with immune-mediated diseases. Patients were receiving treatment with biological medicines. Secondly, to compare those rates with the vaccination percentages of another hospital pharmacy with similar characteristics in which no educational intervention was conducted. METHODS: This was a retrospective cohort study of adult patients with immune-mediated diseases receiving treatment with biological medicines. The medicines were dispensed by the hospital pharmacy departments of two hospitals between 1 January 2019 and 31 December 2020. In Cohort A (intervention group), a health education strategy was implemented with regards to influenza vaccination. Cohort B acted as a control group. The influenza vaccination rates obtained in both cohorts during 2019 and 2020 were compared. RESULT: A total of 355 patients took part in the study - 148 (41.7%) in Cohort A and 207 (58.3%) in Cohort B. The hospital pharmacy department in Cohort A implemented a health education strategy after which the vaccination percentage during the 2020 campaign increased by 38 patients (45.7%), compared with a 10 patient (5.8%) increase in Cohort B (p<0.001). CONCLUSIONS: The health intervention by the pharmacy department had a positive impact. This included an opportunity to improve vaccination take-up and is a strategy to consider when implementing a vaccination programme. Health education is a fundamental objective of healthcare. In our case it led to an increase in vaccination and had a positive impact on public health. It also provides opportunities for pharmacists to work in a multidisciplinary way with other healthcare professionals.

Pan-American Trypanosoma (Megatrypanum) trinaperronei n. sp. in the white-tailed deer Odocoileus virginianus Zimmermann and its deer ked Lipoptena mazamae Rondani, 1878: morphological, developmental and phylogeographical characterisation.

BACKGROUND: The subgenus Megatrypanum Hoare, 1964 of Trypanosoma Gruby, 1843 comprises trypanosomes of cervids and bovids from around the world. Here, the white-tailed deer Odocoileus virginianus (Zimmermann) and its ectoparasite, the deer ked Lipoptena mazamae Rondani, 1878 (hippoboscid fly), were surveyed for trypanosomes in Venezuela. RESULTS: Haemoculturing unveiled 20% infected WTD, while 47% (7/15) of blood samples and 38% (11/29) of ked guts tested positive for the Megatrypanum-specific TthCATL-PCR. CATL and SSU rRNA sequences uncovered a single species of trypanosome. Phylogeny based on SSU rRNA and gGAPDH sequences tightly cluster WTD trypanosomes from Venezuela and the USA, which were strongly supported as geographical variants of the herein described Trypanosoma (Megatrypanum) trinaperronei n. sp. In our analyses, the new species was closest to Trypanosoma sp. D30 from fallow deer (Germany), both nested into TthII alongside other trypanosomes from cervids (North American elk and European fallow, red and sika deer), and bovids (cattle, antelopes and sheep). Insights into the life-cycle of T. trinaperronei n. sp. were obtained from early haemocultures of deer blood and co-culture with mammalian and insect cells showing flagellates resembling Megatrypanum trypanosomes previously reported in deer blood, and deer ked guts. For the first time, a trypanosome from a cervid was cultured and phylogenetically and morphologically (light and electron microscopy) characterised. CONCLUSIONS: In the analyses based on SSU rRNA, gGAPDH, CATL and ITS rDNA sequences, neither cervids nor bovids trypanosomes were monophyletic but intertwined within TthI and TthII major phylogenetic lineages. One host species can harbour more than one species/genotype of trypanosome, but each trypanosome species/genotype was found in a single host species or in phylogenetically closely related hosts. Molecular evidence that L. mazamae may transmit T. trinaperronei n. sp. suggests important evolutionary constraints making tight the tripartite T. trinaperronei-WTD-deer ked association. In a plausible evolutionary scenario, T. trinaperronei n. sp. entered South America with North American white-tailed deer at the Pliocene-Pleistocene boundary following the closure of the Panama Isthmus.

Expanding the pH range of metal-nucleobase complexes for acid-base chemistry: properties of bis(guanine) complexes of (bpy)Pt(II) with either two major or major and minor tautomers bonded simultaneously.

Metal nucleobase complexes provide a potential for acid-base chemistry in the physiological pH range and consequently may contribute or be actively involved in catalytic reactions of nucleic acids, notably of RNAs. Expansion of the available pH range is achieved if additional ligands are involved, for example, an aqua ligand or a second nucleobase, and if relevant pK(a) values are sufficiently close. Two bpy's (bpy = 2,2'-bipyridine) containing Pt(II) complexes have been studied in this context: [Pt(bpy)(9-MeGH-N7)(2)](NO(3))(2) x H(2)O (2) and Pt(bpy)(9-EtG-N7)(9-EtG-N1) x 3 H(2)O (3') (with 9-MeGH = 9-methylguanine; 9-EtGH = 9-ethylguanine). Relevant pK(a) values, as determined by pD-dependent (1)H NMR spectroscopy in D(2)O, of the neutral guanine ligands were found to be ca. 7.78 +/- 0.01 and 8.38 +/- 0.01 for 2, yet 4.00 +/- 0.03 and 7.7 +/- 0.1 for 3' (values converted to H(2)O) for each of the two guanine ligands. These values suggest that complex 3' provides a pH range of roughly 4-8 for potential acid-base chemistry, and furthermore that in favorable cases compounds with two ionizable ligands can function as an acid and a base simultaneously. X-ray crystal structures of both 2 and 3' are presented and, in addition, that of [Pt(bpy)(9-EtGH-N7)(2)](NO(3))(2) x (9-EtGH) x 5 H(2)O (2''). Regarding the use of (1)H NMR spectroscopy for the determination of pK(a) values, we note that chemical shifts referenced to sodium 3-(trimethylsilyl)propanesulfonate must be treated with caution when applying cationic complexes because of the possibility of ion pairing. It can lead to mistakes in chemical shift values.

Prevalence of Salmonella spp. in tonsils, mandibular lymph nodes and faeces of wild boar from Spain and genetic relationship between isolates.

The importance of wild boars as game species in Spain is well known. Their feeding habits and intrusive behaviour, together with a progressively wider spreading of populations, increases the interactions of these animals with livestock and humans. Considering that wild boars could have a potential role in the transmission of certain pathogens as salmonellae, the aims of this study were to determine the prevalence of Salmonella spp. in wild boars hunted in central-western Spain, the occurrence of this pathogen in tonsils, mandibular lymph nodes and faeces (as markers for transmission risk), and to define the phylogenetic relationships among isolated strains, in order to investigate the circulation pathways of bacteria among tissues, animals and estates. Samples from 1,041 hunted wild boars were analysed for the presence of Salmonella spp. by bacteriological culture. Isolates were confirmed by PCR and serotyped in the Spanish national reference laboratory. The genetic relationships between strains were determined by PFGE. The results showed a 7.7% of positive animals (81 wild boars), being tonsils the organ most frequently colonised by Salmonella spp. (18.7%), followed by lymph nodes (5.1%) and faecal samples (2.9%). Serovars Enteritidis and Newport were the most frequent amongst the 34 different serovars obtained. The pulsed-field gel electrophoresis (PGFE) analysis showed a great genetic diversity, with serovars that exhibited different pulsotypes when isolated from different estates and multiple serovars in the same estate. In conclusion, this study reveals the importance of wild boars as carriers and possible transmitters of virulent and/or antimicrobial-resistant clones of Salmonella spp. to livestock and humans.

Outbreaks of antimicrobial resistant Salmonella Choleraesuis in wild boars piglets from central-western Spain.

Salmonella enterica serovar Choleraesuis is the aetiological agent of swine paratyphoid being a highly invasive zoonotic pathogen. Wild boar natural populations are experiencing a demographical expansion as well as some farms are breeding this species to release for hunting with management sometimes identical to that of domestic pigs, including supplementation, grouping, and antibiotic treatments. This situation increases the chance of contact between wild boars and livestock, and potentially induces stress, with different sanitary consequences. The present work aims to describe the clinical features of recent outbreaks caused by S. Choleraesuis in wild boar from central-western Spain, as well as the antimicrobial resistance and phylogenetic relationships of isolates involved. 28 strains of S. Choleraesuis were isolated from 28 different wild boars belonging to 10 different game states located in central western Spain and submitted to the Clinical Veterinary Hospital (CVH) of the University of Extremadura. Samples were taken from different organs and cultured according to the ISO 6579:2002 procedure. Suspicious colonies were identified by PCR and antimicrobial resistance was evaluated by disc diffusion susceptibility test and the presence of the main resistance genes as well as 18 plasmid replicons frequently found among the Enterobacteriaceae was verified by PCR. Pulsed field gel electrophoresis was applied to determine the genetic relationship between isolates. The outbreaks under study were characterized by high mortality (35%-84%) and a septicaemic presentation. S. Choleraesuis was isolated from all the wild boars analysed, and 26 of the 28 isolates presented resistance to at least one antibiotic. The predominant resistances found were against sulphonamide, streptomycin, tetracycline, and doxicicline and sul1, strA-strB, and tetA were the most prevalent resistance genes among isolates. 10 strains carried FIIA, FIB+H/1 or FIIA+H/1 plasmids. PFGE classified the isolates into four different profiles, grouped into two clusters. This results show that prevention against S. Choleraesuis must be considered in the sanitary programs of the wild boar breeders.

Total synthesis of the putative structure of stemonidine: the definitive proof of misassignment.

[structure: see text] The total synthesis of the putative structure of the Stemona alkaloid stemonidine has been completed. The key transformations include a 1,3-dipolar cycloaddition of a chiral nitrone derived from (S)-prolinol and a spirolactonization process involving the generation of the critical stereocenter. The NMR data of the synthetic material do not match those reported for the natural product. It is concluded that the structure assigned to stemonidine is incorrect, and it must be reassigned as stemospironine.

Changes in plasma lipids and increased low-density lipoprotein susceptibility to oxidation in pregnancies complicated by gestational diabetes: consequences of obesity.

Dyslipidemia is associated with increased low-density lipoprotein (LDL) susceptibility to oxidation, a phenomenon associated with endothelial dysfunction, atherosclerosis, cell toxicity, and intrauterine growth retardation. The present study was designed to determine if women developing gestational diabetes mellitus (GDM) have both increased plasma lipids and LDL susceptibility to oxidation throughout pregnancy. We also wanted to study the effects of obesity upon these parameters. A nested case-control study was carried out in 45 women with uncomplicated pregnancies and 62 women diagnosed with GDM following the criteria of the American Diabetes Association. In all women, blood was drawn at 15, 24, and 32 weeks of gestation. Low-density lipoprotein oxidation was initiated by the addition of CuCl2, and formation of conjugated dienes was monitored. Glucose, cholesterol, triglycerides, vitamin E, estradiol, and progesterone were determined. In GDM, elevated levels of glucose, cholesterol, and triglycerides were observed when compared with the control group even in the first trimester, before the detection of diabetes. In the control group, the lag phase in the LDL oxidation was 85.3, 84.4, and 95.6 minutes at 15, 24, and 32 weeks of pregnancy, compared with 63.3, 63.4, and 74.5 minutes in the GDM group (P < .001 in the 3 periods). These differences remained when adjusted for the body mass index. In a multiple linear regression analysis, a negative correlation was observed between the lag phase and the body mass index (P < .001) and cholesterol (P < .001), whereas a positive one appeared with vitamin E (P < .05) and time of gestation (P < .001). In pregnancy, GDM increases LDL susceptibility to oxidation. Obesity and hypercholesterolemia further exacerbate this effect.

Mutagenesis of key amino acids alters activity of a Saccharomyces cerevisiae endo-polygalacturonase expressed in Pichia pastoris.

A polygalacturonase (PG)-encoding gene from Saccharomyces cerevisiae (PGU1) was successfully expressed in the methylotrophic yeast Pichia pastoris. PG secretion was efficiently directed by the S. cerevisiae alpha-factor signal sequence, while the native (PGU1) leader peptide was unable to direct protein export in P. pastoris. The level of PGU1 activity achieved in P. pastoris was significantly enhanced when compared to activity using the same gene in S. cerevisiae. Expression of PG proteins, engineered by site-directed mutagenesis, in P. pastoris showed that aspartic acid residues at positions 179, 200, and 201, and histidine 222 were essential for enzyme activity. Mutation of the two potential glycosylation sites in PGU1 showed that the two residues individually (N318D, N330D) did not affect secreted enzyme activity, but the double mutant caused a 50% reduction in enzyme activity when compared to the wild-type PGU1 transformant.