Epidemiology of Non-Tuberculous Mycobacteria isolated from clinical specimens in Madrid, Spain, from 2013 to 2017.

The epidemiology of non-tuberculous mycobacteria (NTM) in Spain is largely unknown because systematic reporting is not compulsory. The aim of our study was to describe the frequency and diversity of NTM species in our region and their distribution according to the source sample, gender, and age of the patients. We performed a multicenter study of all NTM isolated in 24 public hospitals in Madrid from 2013 to 2017. A total of 6.923 mycobacteria were isolated: 4535 (65.5%) NTM, and 2.388 (34.5%) Mycobacterium tuberculosis complex (MTB). Overall, 61 different NTM species were identified. The most frequently isolated species were Mycobacterium avium complex (47.7%), M. lentiflavum (12.2%), M. gordonae (9.2%), M. fortuitum (8.9%), and M. abscessus (3.9%). Whereas MTB cases were stable during the study period, the number of NTM isolates increased considerably from 930 isolates in 2013 to 1012 in 2017; a sharp increase occurred in the last year. The rise in NTM isolates was mostly due to M. lentiflavum, M. kansasii, and M. abscessus mainly isolated from respiratory specimens in patients older than 60. The increase in isolation rate of NTM in our region is consistent with the increasing rates reported worldwide in the last decades. The rise in NTM isolates was mainly attributed to M. lentiflavum but it also should be noted the increasing of species with high pathogenic potential such as M. kansasii and M. abscessus.

Practical application of brief cognitive tests.

INTRODUCTION: Brief cognitive tests (BCT) may help detect cognitive impairment (CI) in the clinical setting. Several BCT have been developed and/or validated in our country, but we lack specific recommendations for use. DEVELOPMENT: Review of studies on the diagnostic accuracy of BCT for CI, using studies conducted in Spain with BCT which take less than 20 min. We provide recommendations of use based on expert consensus and established on the basis of BCT characteristics and study results. CONCLUSION: The Fototest, the Memory Impairment Screen (MIS) and the Mini-Mental State Examination (MMSE) are the preferred options in primary care; other BCT (Clock Drawing Test [CDT], test of verbal fluency [TVF]) may also be administered in cases of negative results with persistent suspected CI or concern (stepwise approach). In the specialised care setting, a systematic assessment of the different cognitive domains should be conducted using the Montreal Cognitive Assessment, the MMSE, the Rowland Universal Dementia Assessment, the Addenbrooke's Cognitive Examination, or by means of a stepwise or combined approach involving more simple tests (CDT, TVF, Fototest, MIS, Memory Alteration Test, Eurotest). Associating an informant questionnaire (IQ) with the BCT is superior to the BCT alone for the detection of CI. The choice of instruments will depend on the patient's characteristics, the clinician's experience, and available time. The BCT and IQ must reinforce - but never substitute - clinical judgment, patient-doctor communication, and inter-professional dialogue.

Robust optimization of alginate-Carbopol 940 bead formulations.

Formulation process is a very complex activity which sometimes implicates taking decisions about parameters or variables to obtain the best results in a high variability or uncertainty context. Therefore, robust optimization tools can be very useful for obtaining high quality formulations. This paper proposes the optimization of different responses through the robust Taguchi method. Each response was evaluated like a noise variable, allowing the application of Taguchi techniques to obtain a response under the point of view of the signal to noise ratio. A L(18) Taguchi orthogonal array design was employed to investigate the effect of eight independent variables involved in the formulation of alginate-Carbopol beads. Responses evaluated were related to drug release profile from beads (t(50%) and AUC), swelling performance, encapsulation efficiency, shape and size parameters. Confirmation tests to verify the prediction model were carried out and the obtained results were very similar to those predicted in every profile. Results reveal that the robust optimization is a very useful approach that allows greater precision and accuracy to the desired value.

Amino acids and volatile compounds in wines from Cabernet Sauvignon and Tempranillo varieties subjected to malolactic fermentation in barrels.

The aim of the present paper is to compare the behaviour of industrial lactic bacteria and indigenous bacteria of the cellar when malolactic fermentation was carried out in barrels. The effects of these bacteria on the concentration of metabolised amino acids during malolactic fermentation and on the composition of volatile compounds both before and after malolactic fermentation are studied. The experiment was performed with wines of the Tempranillo and Cabernet Sauvignon varieties. An analysis has been made of the easily extractable volatile compounds of the wood and the compounds from the grapes, and the action of the yeasts during the alcoholic fermentation. Acetoin and diacetyl decreased during the malolactic fermentation in barrels and the concentrations of furfural and its derivatives were up to 100 times higher in wines not subjected to malolactic fermentation. Most of the volatile phenols increased during the malolactic fermentation in wines of the Tempranillo variety, while only guaiacol (p < 0.05) and t-isoeugenol increased in the Cabernet Sauvignon wines. The decrease in amino acids during the malolactic fermentation depends much more on the variety than on the bacterial strain which carries out the malolactic fermentation.

Impact of the COVID-19 pandemic on tuberculosis management in Spain.

BACKGROUND: The impact of COVID-19 on the diagnosis and management of tuberculosis (TB) patients is unknown. METHODS: Participating centres completed a structured web-based survey regarding changes to TB patient management during the COVID-19 pandemic. The study also included data from participating centres on patients aged ≥18 diagnosed with TB in 2 periods: March 15 to June 30, 2020 and March 15 to June 30, 2019. Clinical variables and information about patient household contacts were retrospectively collected. RESULTS: A total of 7 (70%) TB units reported changes in their usual TB team operations. Across both periods of study, 169 patients were diagnosed with active TB (90 in 2019, 79 in 2020). Patients diagnosed in 2020 showed more frequent bilateral lesions in chest X-ray than patients diagnosed in 2019 (P = 0.004). There was a higher percentage of latent TB infection and active TB among children in households of patients diagnosed in 2020, compared with 2019 (P = 0.001). CONCLUSIONS: The COVID-19 pandemic has caused substantial changes in TB care. TB patients diagnosed during the COVID-19 pandemic showed more extended pulmonary forms. The increase in latent TB infection and active TB in children of patient households could reflect increased household transmission due to anti-COVID-19 measures.

Attitude and knowledge of hearing loss among medical doctors selected to initiate a residency in Mexico.

Early diagnosis and intervention of hearing loss are directly influenced by the knowledge and attitude towards this condition among medical personnel, particularly in countries where screening is not performed routinely. The objective of this paper was to evaluate the attitude and knowledge of hearing loss in a group of physicians. A questionnaire with five Likert-type items and five multiple choice and fill-in-the-blank items was completed by 2727 physicians selected to start a medical residency. Results suggested that physicians' knowledge level on the matter is deficient and their attitude is far from the ideal; however, physicians selected for a residency in audiology showed slightly better results.

A simple device for headspace sorptive extraction prior to gas chromatography-mass spectrometry analysis.

A device for headspace sorptive extraction (HSSE) combined with gas chromatography separation and mass spectrometry detection (GC-MS) is presented. The gadget is based on a simple magnetic disk that permits the stir bar to be placed at the top of the sample vial. The complete surface of the coated stir bar is exposed to the headspace atmosphere, and the movement of the bar caused by the magnetic stirrer in the liquid sample facilitates the sorption process. The final consequence is increased sensitivity and repeatability when compared with a glass-made comercial device for the same purpose. Successful determination of ten chlorobenzenes in waters testifies the good performance of the new device.

High-solids anaerobic digestion: comparison of three pilot scales.

Two experiments were undertaken in three different experimental set-ups in order to compare them: an industrial 21-m3 pilot reactor, a new 40-l laboratory pilot reactor and bmp type plasma bottles. Three consecutive batch dry digestion tests of municipal solid waste were performed under mesophilic conditions with the same feedstock in all vessels. Biogas and methane production at the end of the tests were similar (around 200 m3 CH4STP/tVS) for both pilot reactors and were different from the bottle tests. The dynamics of methane production and VFA accumulation concurred. However, the maximal levels of VFA transitory accumulation varied between reactors and between runs in a same reactor. Ammonia levels were similar in both reactors. These results show that the new reactor accurately imitates the conditions found in the larger one. Adaptation of microorganisms to the waste and operating conditions was also pointed out along the consecutive batches. Thermophilic semi-continuous tests were performed in both reactors with similar conditions. The methane production efficiencies were similar.

In situ ionic liquid dispersive liquid-liquid microextraction coupled to gas chromatography-mass spectrometry for the determination of organophosphorus pesticides.

Nine organophosphorus pesticides (OPPs) were determined in environmental waters from different origins using in situ ionic liquid dispersive liquid microextraction (IL-DLLME). This preconcentration technique was coupled to gas chromatography-mass spectrometry (GC-MS) using microvial insert thermal desorption, an approach that uses a thermal desorption injector as sample introduction system. The parameters affecting both the microextraction and sample injection steps were optimized. The proposed method showed good precision, with RSD values ranging from 4.1 to 9.7%, accuracy with recoveries in the 85-118% range, and sensitivity with DLs ranging from 5 to 16 ng L-1.

Factors affecting the incidence of contrast-induced nephropathy in patients undergoing computed tomography. / Incidencia de nefropatía por contraste en pacientes sometidos a tomografía computarizada: ¿qué factores la condicionan?

OBJECTIVE: To analyze the incidence of contrast-induced nephropathy in a cohort of patients undergoing computed tomography (CT) with intravenous iodinated contrast material. To evaluate the efficacy of N-acetylcysteine in preventing contrast-induced nephropathy. PATIENTS AND METHODS: This prospective observational study was carried out in the months comprising March 2016 through July 2016. We selected the first five patients scheduled to undergo CT examination each day who agreed to participate and signed the informed consent form. We recorded patients' cardiovascular histories, chronic treatments, and indications for the CT examination. We measured blood levels of creatinine and urea before and after the CT examination. We used the Modification of Diet in Renal Disease (MDRD-4) equation to estimate the glomerular filtration rate. We analyzed the type and dose of contrast material. We recorded whether N-acetylcysteine was administered before the CT examination. We used SPSS 15.0® to compare means and proportions. Statistical significance was set at p < 0.05. RESULTS: No incidents of contrast-induced nephropathy were detected in any of the 202 patients included [mean age, 63.92 ± 12 years (range 22-87); 57.4% male; 21.8% diabetic; 39.6% hypertensive; 87.1% had MDRD4 ≥ 60 ml/min/1.73 m2 (89.45 ± 14, range 62.36-134.14) and 12.9% had MDRD4 < 60 ml/min/1.73 m2 (45.38 ± 11, range 9.16-58.90)]. The most common indication for CT examinations was oncologic (81.2%). The only contrast agent administered was iopamidol; the mean dose was 107.83 ± 11 ml (range 70-140). The mean interval between pre-CT and post-CT laboratory tests was 4.06 ± 1 days. Only 13 patients received N-acetylcysteine; 9 of these had MDRD < 60 ml/min/1.73 m2 and 4 had MDRD4 ≥ 60 ml/min/1.73 m2 (p = 0.000). CONCLUSIONS: The incidence of contrast-induced nephropathy was not significant in patients with glomerular filtration rates greater than 30 ml/min/1.73 m2: these favorable results might be due to analyzing only scheduled examinations and to using relatively low doses of a "nonionic" iodinated contrast agent.

Activity of ertapenem and other antimicrobials against ESBL-producing enterobacteria isolated from urine in patients from Madrid.

The objective of this study was to evaluate the activity of ertapenem and other antimicrobials against extended-spectrum beta-lactamase (ESBL)-producing enterobacteria isolated from patients' urine samples at 4 community healthcare centers in the Madrid (Spain) area and to determine the prevalence of ESBL-producing enterobacteria in community-acquired urinary tract infections. The antibiotic susceptibility results were compared by patient age and sex. A total of 293 strains were studied. The minimum inhibitory concentration (MIC) for each antibiotic was determined using the agar dilution method. The tested carbapenems were the antibiotics with the greatest activity (ertapenem MIC(90)=0.06 mg/l; imipenem MIC(90)=0.5 mg/l), with no intermediate or resistant strains being observed. High rates of resistance to ciprofloxacin (80.9%) and cotrimoxazole were observed (62.1%). The global prevalence of ESBL-producing enterobacteria was 3.6% (293/8,139). Prevalence according to areas was 5.3% in Getafe, 3.45% in Arguelles, 3.02% in Alcala de Henares and 3.56% in Mostoles. The global prevalence of ESBL-producing Escherichia coli was 4.15% (279/6,721). The analysis of resistance according to patient sex (males versus females) showed no significant differences. The analysis of resistance according to patient age (<50 years versus > or = 50 years) showed statistically significant differences (more resistance among subjects > or = 50 years old) for cotrimoxazole (OR=0.43, 95%CI: 0.20-0.93, p=0.018) and ciprofloxacin (OR=0.32, 95%CI: 0.14-0.74, p=0.0027). In view of the good activity shown by ertapenem, and the continuous increase in the prevalence of ESBL strains, this antibiotic and some of the others could be a good choice for the treatment of community-acquired urinary tract infections produced by such bacteria in Spain.

Multidimensional chromatographic approach applied to the identification of novel aroma compounds in wine. Identification of ethyl cyclohexanoate, ethyl 2-hydroxy-3-methylbutyrate and ethyl 2-hydroxy-4-methylpentanoate.

A multidimensional chromatographic strategy has been developed and optimized with the purpose of identifying different odorants potentially relevant to the aroma and flavor of aged wines from Madeira or Sherry. Different techniques of extraction and fractionation were studied in order to get clear olfactometric and spectrometric signals from the target odorants. The best results were obtained with a dynamic headspace extraction followed by a fractionation on a normal phase medium pressure liquid chromatography on a silicagel column. Large volumes (50 microl) of the concentrated fractions were further analyzed in a dual gas chromatography-mass spectrometric system (GC-MS) equipped with two olfactometric ports. The strategy made it possible to identify in wine by first time the presence of the powerful strawberry-smelling compound, ethyl cyclohexanoate, and of two other novel fruity esters, ethyl 2-hydroxy-3-methylbutyrate and ethyl 2-hydroxy-4-methylpentanoate. Some other unidentified odorants could be isolated and their mass spectra are given.

Direct sample introduction-gas chromatography-mass spectrometry for the determination of haloanisole compounds in cork stoppers.

A solventless analytical method is proposed for analyzing the compounds responsible for cork taint in cork stoppers. Direct sample introduction (DSI) is evaluated as a sample introduction system for the gas chromatography-mass spectrometry (GC-MS) determination of four haloanisoles (HAs) in cork samples. Several parameters affecting the DSI step, including desorption temperature and time, gas flow rate and other focusing parameters, were optimized using univariate and multivariate approaches. The proposed method shows high sensitivity and minimises sample handling, with detection limits of 1.6-2.6ngg-1, depending on the compound. The suitability of the optimized procedure as a screening method was evaluated by obtaining decision limits (CCα) and detection capabilities (CCß) for each analyte, which were found to be in 6.9-11.8 and 8.7-14.8ngg-1, respectively, depending on the compound. Twenty-four cork samples were analysed, and 2,4,6-trichloroanisole was found in four of them at levels between 12.6 and 53ngg-1.

Dispersive liquid-liquid microextraction for the determination of nitrophenols in soils by microvial insert large volume injection-gas chromatography-mass spectrometry.

A rapid and sensitive procedure for the determination of six NPs in soils by gas chromatography and mass spectrometry (GC-MS) is proposed. Ultrasound assisted extraction (UAE) is used for NP extraction from soil matrices to an organic solvent, while the environmentally friendly technique dispersive liquid-liquid microextraction (DLLME) is used for the preconcentration of the resulting UAE extracts. NPs were derivatized by applying an "in-situ" acetylation procedure, before being injected into the GC-MS system using microvial insert large volume injection (LVI). Several parameters affecting UAE, DLLME, derivatization and injection steps were investigated. The optimized procedure provided recoveries of 86-111% from spiked samples. Precision values of the procedure (expressed as relative standard deviation, RSD) lower than 12%, and limits of quantification ranging from 1.3 to 2.6ngg(-1), depending on the compound, were obtained. Twenty soil samples, obtained from military, industrial and agricultural areas, were analyzed by the proposed method. Two of the analytes were quantified in two of the samples obtained from industrial areas, at concentrations in the 4.8-9.6ngg(-1) range.

Cloud point extraction and gas chromatography with direct microvial insert thermal desorption for the determination of haloanisoles in alcoholic beverages.

A sensitive analytical procedure for the determination of four haloanisoles (2,4,6 trichloroanisole, 2,4,6-tribromoanisole, 2,3,4,6-tetrachloroanisole and pentachloroanisole) related with cork taint defects in wines, in different types of alcoholic beverages has been developed. The analytes were extracted from the matrix samples by cloud point extraction (CPE) using Triton X-114 heated to 75°C, and the surfactant rich phase was separated by centrifugation. By means of direct microvial insert thermal desorption, 20µL of the CPE obtained extract was submitted to gas chromatography-mass spectrometry (GC-MS) analysis. The parameters affecting the CPE and microvial insert thermal desorption were optimized. Quantification was carried by matrix-matched calibration using an internal standard. Detection limits ranged between 12.9 and 20.8ngL(-1), depending on the compound, for beer and wine samples, whereas for whiskies values in the 46.3-48ngL(-1) range were obtained, since these samples were diluted for analysis. Recoveries for alcoholic beverages were in the 89-111% range, depending on the analyte and the sample.

Evaluation of three headspace sorptive extraction coatings for the determination of volatile terpenes in honey using gas chromatography-mass spectrometry.

Headspace sorptive extraction (HSSE) was used to preconcentrate seven monoterpenes (eucalyptol, linalool, menthol, geraniol, carvacrol, thymol and eugenol) for separation by gas chromatography and mass spectrometry (GC-MS). Three commercially available coatings for the stir bars, namely Polydimethylsiloxane (PDMS), polyacrilate (PA) and Ethylene glycol-silicone (EG-Silicone), were tested, and the influential parameters both in the adsorption and the thermal desorption steps were optimized. PDMS provided the best sensitivity for linalool, geraniol, menthol and eucalyptol, whereas EG-Silicone was best for extracting the phenolic monoterpenes studied. Considering the average obtained slopes from all compounds, PDMS pointed as the best option, and the analytical characteristics for the HSSE-TD-GC-MS method using this coating were obtained. Quantification of the samples was carried out by matrix-matched calibration using a synthetic honey. Detection limits ranged between 0.007 and 0.032 ng g(-1), depending on the compound. Twelve honey samples of different floral origins were analyzed using the HSSE-GC-MS method, the analytes being detected at concentrations up to 64 ng g(-1).

Direct sample introduction gas chromatography and mass spectrometry for the determination of phthalate esters in cleaning products.

A method using direct sample introduction (DSI) coupled to gas chromatography-mass spectrometry (GC-MS) is developed for the determination of six phthalate esters (dimethyl, diethyl, dibutyl, butylbenzyl, diethylhexyl and dioctyl phthalate) in cleaning products. The different variables involved in the DSI step, including venting time and temperature, vaporisation time and temperature, injector heating temperature and gas flow rate and pressure, were evaluated and optimised using Taguchi orthogonal arrays. The proposed method, using calibration against methanolic standards, showed good linearity in the 0.05-15 µg g(-1) range and good repeatability, with RSD values ranging from 3.5% to 5.7%. Quantification limits between 0.010 and 0.041 µg g(-1), depending on the compound, were attained, while recovery assays provided values from 83% to 115%. Twenty-seven cleaning products were analysed using the DSI-GC-MS method, being four phthalates (dimethyl, diethyl, dibutyl and diethylhexyl phthalate) found in fourteen of them at concentration levels in the 0.1-21 µg g(-1) range. Compared with the most common GC injection technique, which uses the split/splitless injector, the proposed DSI procedure provided larger peak areas and lower detection limits, as result of the greater injected volume and reduction in noise.

Changes in dietary fatty acids modify the decreased lipolytic beta3-adrenergic response to hyperinsulinemia in adipocytes from pregnant and nonpregnant rats.

The effect of dietary olive oil and fish oil on the lipolytic dose-response of the beta3-adrenergic agonists, epinephrine, isoproterenol, BRL-37344, and CGP-12177, in adipocytes was studied in pregnant and virgin rats either untreated or under hyperinsulinemic-euglycemic conditions. Rats were fed a semisynthetic diet containing 5% of either olive oil or fish oil and studied at day 20 of treatment and/or gestation. Plasma glucose was lower and plasma insulin, triglycerides, and free fatty acids (FFAs) were higher in pregnant versus virgin rats, and the insulin sensitivity index was lower in the former. Lumbar adipose tissue phospholipid fatty acids showed a significantly higher monounsaturated fatty acid and a lower (n - 3) fatty acid content in rats fed the olive oil diet versus the fish oil diet. The lipolytic dose-response curve of either adrenergic agent was always lower in adipocytes from untreated pregnant rats versus virgin rats, and whereas the hyperinsulinemic-euglycemic clamp decreased these responses in adipocytes from virgin rats fed the olive oil diet only, adipocytes from pregnant rats always showed a decreased dose-response lipolytic curve. Thus, the lipolytic responsiveness of beta3-adrenoceptor (beta3-AR) agonists by adipocytes is impaired in cells from rats made hyperinsulinemic chronically by pregnancy or acutely by the hyperinsulinemic-euglycemic clamp, but such response to the acute condition disappears when the adipocyte phospholipid composition is modified by changes in dietary fatty acids.

Fast analysis of important wine volatile compounds development and validation of a new method based on gas chromatographic-flame ionisation detection analysis of dichloromethane microextracts.

A method for the simultaneous determination of major (10-200 mg/l) and minor (0.1-10 mg/l) volatile compounds from wine has been optimised and validated. A 3-ml volume of wine is diluted with water (7 ml), salted with 4.5 g of ammonium sulfate and extracted with 0.2 ml of dichloromethane. The extract is injected in the split mode in a GC system, separated on a Carbowax 20M capillary column and detected by flame ionisation detection. Volatiles from wine are divided into four groups according to their behaviour in the extraction, and a specific internal standard has been selected for each group. The method allows satisfactory determination of more than 30 volatile compounds of wine. Compounds analysed include acetaldehyde, diacetyl, acetoine (3-hydroxy butanone), fusel alcohols and their acetates, and fatty acids and their ethyl esters. The linear dynamic range of the method covers the normal range of occurrence of analytes in wine and extends from at least one magnitude order to more than two, with typical r2 between 0.9938 and 0.9998. Reproducibility ranges from 3.1 to 10% (as RSD) with 5.5% as the average. The analysis of spiked samples has shown that matrix effects do not significantly affect method performance.

Use of solid-liquid distribution coefficients to determine retention properties of Porapak-Q resins. Determination of optimal conditions to isolate alkyl-methoxypyrazines and beta-damascenone from wine.

The solid-liquid distribution coefficients of different analytes--all of which are important aroma compounds--between hydroalcoholic solutions or wines and different sorbents have been determined by measuring the amount of analyte removed by a given mass of sorbent in equilibrium with a given volume of standard solution. These data have shown that the best extraction conditions for non-polar compounds from wine are the use of Porapak-Q resins and 6% (v/v) alcoholic solutions. Phase ratio, hold-up volumes and number of plates for Porapak-Q beds have been measured in different experiments. With all these data it has been possible to calculate breakthrough volumes in good agreement with experimental results. The Lövkvist-Jönsson model is more appropriate for estimating breakthrough volumes of a 2-cm Porapak-Q bed. The model estimates that a 5-cm bed is needed to achieve a quantitative recovery of 3-alkyl-2-methoxypyrazines and beta-damascenone from 500 ml of wine (diluted to 1000 ml with water). Experimental results confirm the predictions of the model and show that in a single isolation step detection limits below 10 ng/l can be reached for these compounds using GC-MS detection.