Exploring the root canal microbiome in previously treated teeth: A comparative study of diversity and metabolic pathways across two geographical locations.

AIM: To analyse and compare the root canal microbiome present in root-filled teeth of two different geographical populations, and to study their functional potential using a next-generation sequencing approach. METHODOLOGY: Sequencing data obtained from surgical specimens from previously treated teeth with periapical bone loss from Spain and USA were included in the study. Taxa were classified using SILVA v.138 database. Differences in genera abundances among the 10 most abundant genera were evaluated using a Kruskal-Wallis test. Alpha diversity indices were calculated in mothur. The Shannon and Chao1 indices were used. Analyses of similarity (ANOSIM) to determine differences in community composition were done in mothur, with Bonferroni correction for multiple comparisons. p-Values < .05 were considered statistically significant. Identification of enriched bacteria function prediction in the study groups (KEGG pathways) was carried out by linear discriminant analysis effect size (LEfSe) via Python 3.7.6. RESULTS: A greater alpha-diversity (Shannon and Chao1 indices) was observed from samples obtained in Spain (p = .002). Geography showed no significant effects on community composition via an ANOSIM using Bray-Curtis dissimilarities (R = 0.03, p = .21). Bacterial functional analysis prediction obtained by PICRUSt showed that 5.7% KEGG pathways differed between the Spain and US samples. CONCLUSIONS: The taxonomic assessment alone does not fully capture the microbiome's differences from two different geographical locations. Carbohydrate and amino acid metabolism were enriched in samples from Spain, while samples from USA had a higher representation of pathways related to nitrogen, propanoate metabolism, and secretion systems.

Validation of the Unesp-Botucatu pig composite acute pain scale (UPAPS) in piglets undergoing castration.

To accurately assess pain and support broadly-based analgesic protocols to mitigate swine pain, it is imperative to develop and validate a species-specific pain scale. The objective of this study was to investigate the clinical validity and reliability of an acute pain scale (UPAPS) adapted for newborn piglets undergoing castration. Thirty-nine male piglets (five days of age, 1.62 ± 0.23 kg BW) served as their own control, were enrolled in the study and underwent castration in conjunction with an injectable analgesic administered one-hour post-castration (flunixin meglumine 2.2 mg/kg IM). An additional 10, non-painful female piglets were included to account for the effect of natural behavioral variation by day on pain scale results. Behavior of each piglet was video recorded continuously at four recording periods (24 h pre-castration, 15 min post-castration, 3 and 24 h post-castration). Pre- and post-operative pain was assessed by using a 4-point scale (score 0-3) including the following six behavioral items: posture, interaction and interest in surroundings, activity, attention to the affected area, nursing, and miscellaneous behavior. Behavior was assessed by two trained blinded observers and statistical analysis was performed using R software. Inter-observer agreement was very good (ICC = 0.81). The scale was unidimensional based on the principal component analysis, all items except for nursing were representative (rs ≥ 0.74) and had excellent internal consistency (Cronbach's alpha ≥ 0.85). The sum of scores were higher in castrated piglets post-procedure compared to pre-procedure, and higher than in non-painful female piglets confirming responsiveness and construct validity, respectively. Scale sensitivity was good when piglets were awake (92.9%) and specificity was moderate (78.6%). The scale had excellent discriminatory ability (area under the curve > 0.92) and the optimal cut-off sum for analgesia was 4 out of 15. The UPAPS scale is a valid and reliable clinical tool to assess acute pain in castrated pre-weaned piglets.

γ-Cyclodextrin-graphene quantum dots-chitosan modified screen-printed electrode for sensing of fluoroquinolones.

An innovative electrochemical approach based on screen-printed carbon electrodes (SPCEs) modified with graphene quantum dots (GQDs) functionalized with γ-cyclodextrin (γ-CD) and assembled to chitosan (CHI) is designed for the assessment of the total content of fluoroquinolones (FQs) in animal source products. For the design of the bionanocomposite, carboxylated graphene quantum dots synthesized from uric acid as precursor were functionalized with γ-CD using succinic acid as a linker. Physic-chemical and nanostructural characterization of the ensuing nanoparticles was performed by high-resolution transmission scanning microscopy, dynamic light scattering, Z potential measurement, Fourier transformed infrared spectroscopy and X-ray diffraction. Electrochemical properties of assembled bionanocomposite like potential difference, kinetic electronic transfer constant and electroactive area among other parameters were assessed by cyclic voltammetry and differential pulse voltammetry using potassium ferricyanide as redox probe. The oxidation behaviour of four representative quinolones with distinctive structures was studied, obtaining in all cases the same number of involved e- (2) and H+ (2) in their oxidation. These results led us to propose a single and consistent oxidation mechanism for all the checked analytes. The γ-CD-GQDs-CHI/SPCE sensor displayed a boosted electroanalytical performance in terms of linear range (4-250 µM), sensibility (LOD = 1.2 µM) and selectivity. This electrochemical strategy allowed the determination of FQs total amount in complex processed food like broths, bouillon cubes and milkshakes at three concentration levels (150, 75 and 37.5 µM) for both equimolar and different ratio FQs mixtures with recovery values ranging from 90 to 106%.

[Recommendations for the use of neurophysiological techniques in the diagnosis of brain death from the Clinical Neurophysiology Society of the Communities of Valencia and Murcia]. / Recomendaciones para el empleo de técnicas neurofisiológicas en el diagnóstico de muerte encefálica de la Sociedad de Neurofisiología Clínica de las Comunidades de Valencia y Murcia.

In Spain organ transplantation constitutes one of the greatest challenges and teamwork of hospital centres. It is estimated that in the year 2020 Spain contributed 19% of all donors to the European Union. The confirmatory support diagnosis recommends by law some complementary techniques in certain cases, including neurophysiological techniques, especially the use of electroencephalogram and evoked potentials. These cases require the clinical neurophysiologist to make the right clinical and technical decisions for the correct performance and interpretation of the same. To date, there is no national consensus on the performance of these techniques. Updated bibliographic review on neurophysiological techniques (electroencephalogram and evoked potentials). Analysis by Delphi method and expert judgment of the working group of the Clinical Neurophysiology Society of the Communities of Valencia and Murcia. Neurophysiological techniques can be a support in the diagnosis of encephalic death, both confirmatory and to shorten observation times. In order to perform them, minimum technical standards are required to allow optimal performance of the studies, especially taking into account medication, hemodynamic situation, absence of hypothermia, and age group. We present the first guide in Spanish elaborated by the Society of Neurophysiology of the Communities of Valencia and Murcia for the performance in our hospitals of neurophysiological techniques in the diagnosis of brain death.

TITLE: Recomendaciones para el empleo de técnicas neurofisiológicas en el diagnóstico de muerte encefálica de la Sociedad de Neurofisiología Clínica de las Comunidades de Valencia y Murcia.En España, el trasplante de órganos constituye uno de los mayores retos y trabajo en equipo de los centros hospitalarios. Se estima que en 2020 España aportó a la Unión Europea el 19% de la totalidad de los donantes. El diagnóstico de apoyo confirmatorio recomienda por ley algunas técnicas complementarias en determinados casos, entre ellas las técnicas neurofisiológicas, en especial el uso del electroencefalograma y los potenciales evocados. Estos casos plantean al neurofisiólogo clínico la toma acertada de decisiones tanto clínicas como técnicas para su correcta realización e interpretación. Hasta ahora no existe a nivel nacional un consenso de realización de estas técnicas. Es una revisión bibliográfica actualizada sobre las técnicas neurofisiológicas (electroencefalograma y potenciales evocados), con análisis mediante método Delphi y juicio de expertos del grupo de trabajo de la Sociedad de Neurofisiología Clínica de las Comunidades de Valencia y Murcia. Las técnicas neurofisiológicas permiten ser un apoyo en el diagnóstico de muerte encefálica, tanto de forma confirmatoria como para acortar tiempos de observación. Para su realización se precisan unos mínimos estándares técnicos que permitan realizar de forma óptima los estudios. Especialmente hay que tener en cuenta la medicación, la situación hemodinámica, la ausencia de hipotermia y el grupo de edad. Presentamos la primera guía en castellano elaborada por la Sociedad de Neurofisiología de las Comunidades de Valencia y Murcia para la realización en nuestros hospitales de las técnicas neurofisiológicas en el diagnóstico de muerte encefálica.

Natural and Anthropogenic Processes Affecting Domestic Groundwater Quality within the Northwestern Appalachian Basin.

Domestic wells serve as the primary drinking-water source for rural residents in the northern Appalachian Basin (NAB), despite a limited understanding of contaminant distributions in groundwater sources. We employ a newly collected dataset of 216 water samples from domestic wells in Ohio and West Virginia and an integrated contaminant-source attribution method to describe water quality in the western NAB and characterize key agents influencing contaminant distributions. Our results reveal arsenic and nitrate concentrations above federal maximum contaminant levels (MCLs) in 6.8 and 1.3% of samples and manganese concentrations above health advisory in 7.3% of samples. Recently recharged groundwaters beneath upland regions appear vulnerable to surface-related impacts, including nitrate pollution from agricultural activities and salinization from road salting and domestic sewage sources. Valley regions serve as terminal discharge points for long-residence-time groundwaters, where mixing with basin brines is possible. Arsenic impairments occurred in alkaline groundwaters with major-ion compositions altered by ion exchange and in low-oxygen metal-rich groundwaters. Mixing with as much as 4-10% of mine discharge-like waters was observed near coal mining operations. Our study provides new insights into key agents of groundwater impairment in an understudied region of the NAB and presents an integrated approach for contaminant-source attribution applicable to other regions of intensive resource extraction.

Subconjunctival human dirofilariasis by Dirofilaria repens in the Mediterranean Basin.

Purpose: Human dirofilariasis cases have increased in Europe for the last few decades. We document an autochthonous case of dirofilariasis infection in the subconjunctival space of a 60-year-old Spanish man. Observations: The patient presented with pain in his right eye, which was diagnosed as epi-scleritis. In spite of the treatment administered, no improvement was reported. An external exam showed conjunctival congestion on the temporal part of the right eye bulbar conjunctiva. The anterior chamber showed no flare or cells in both eyes and dilated fundus exam was normal. Nevertheless, a long whitish vermiform mobile mass was detected under the conjunctiva in the temporal part of the right eye. The worm was surgically removed and diagnosed morphologically and also molecularly as Dirofilaria repens. This species uses dogs and wild canids as definitive hosts and principal reservoirs, while mosquito species are transmitters. Humans act as accidental hosts, and clinical manifestations depend on the location of the worm in the organs and tissues. Conclusions and importance: The correct diagnosis and control of subcutaneous/ocular dirofilariasis by D. repens require several professionals under the One Health approach to deal with this zoonotic disease, which poses a serious public health problem, at least in the Mediterranean Basin.

Difficult intravenous access in a paediatric intensive care unit.

INTRODUCTION: Multiple attempts during peripheral cannulation can have major consequences for patients, relatives, and healthcare professionals, therefore we set out to determine the extent of this problem in a paediatric intensive care unit (PICU). OBJECTIVES: The main aim was to describe peripheral venous catheter (PVC) and peripherally inserted central catheter (PICC) cannulation in children in the PICU. Secondary objectives were to determine the success rate of the first cannulation attempt, to quantify patients with difficult venous access (DVA), and to explore the association between DVA and sociodemographic, technique and nursing-related characteristics. METHOD: A cross-sectional descriptive study. Consecutive sampling was used to recruit patients aged 0-18 years admitted to the PICU who required peripheral venous cannulation. An ad hoc questionnaire was used for this purpose, including the presence of DVA as an independent variable. RESULTS: A total of 163 venous cannulations were reported. A total of 55.8% (91) were performed in patients under 1 year of age. Of these, 38.7% (63) were successful on the first attempt and 36.8% (60) had DVA. When there was DVA, 85% (51) of patients had complications, median time to cannulation by short CVP was 30 minutes [15-53] and 2 or more nurses were required on 80% (48) of occasions. CONCLUSIONS: We found a low success rate at first attempt and a high proportion of DVA. More nurses and time were employed during cannulation and complications increased if the patient had DVA. A statistically significant association was found between DVA and age, weight, poor perfusion, veins that were neither visible nor palpable, DIVA score ≥ 4, history of difficult intravenous access, complications, number of nurses and time spent.

Cefiderocol.

Cefiderocol is a new siderophore cephalosporin with potent in vitro activity against gram-negative bacilli including Enterobacterales that produce all kinds of carbapenemases and non-fermenting Gram-negative with difficult-to-treat resistance. As a ß-lactam, its efficacy is optimized in extended-perfusion and requires dose adjustment in renal dysfunction and hyperclearance. Its efficacy has been validated in three clinical trials, one of them in the treatment of hospital-acquired pneumonia and ventilator-associated pneumonia. The clinical trial aimed at difficult-to-treat gram-negatives achieved the clinical and microbiological target, but the increase in mortality observed in the cefiderocol arm makes it necessary to demonstrate efficacy in real clinical practice. Cefiderocol is a good option among the new ß-lactams for the treatment of pneumonia caused by Gram-negative bacilli carbapenem-resistant.

Cyclodextrin-modified graphene quantum dots as a novel additive for the selective separation of bioactive compounds by capillary electrophoresis.

Highly reliable separation and determination of various biologically active compounds were achieved using capillary electrophoresis (CE) based on ß-cyclodextrin-functionalized graphene quantum dots (ßcd-GQDs) as the background electrolyte additive. ßcd-GQDs improve the separation efficiency between peaks of all analytes. No addition of surfactants or organic solvents was needed in the running buffer containing ßcd-GQDs. Up to eight consecutive runs were acquired with high precision for the separation of resveratrol, pyridoxine, riboflavin, catechin, ascorbic acid, quercetin, curcumin, and even of several of their structural analogs. Baseline separation was achieved within just 13 min as a result of the effective mobility of the analytes along the capillary owing to the differential interaction with the additive. The proposed analytical method displayed a good resolution of peaks for all species selecting two absorption wavelengths in the diode array detector. Detection limits lower than 0.28 µg mL-1 were found for all compounds and precision values were in the range of 2.1-4.0% in terms of the peak area of the analytes. The usefulness of the GQD-assisted selectivity-enhanced CE method was verified by the analysis of food and dietary supplements. The applicability to such complex matrices and the easy and low-cost GQD preparation open the door for routine analyses of food and natural products. The concept of using such a dual approach (macromolecules and nanotechnology) has been explored to tackle the separation of various bioactive compounds in nutritional supplements and food. Schematic illustration of the electrophoretic separation of the bioactive molecules in the capillary which is filled with the running solution without (top) and with ßcd-GQDs (bottom). The fused silica capillary with negatively ionizable silanol groups at the wall. The voltage is applied at positive polarity at the outlet. R, riboflavin; r, resveratrol; P, pyridoxine; C, catechin; c, curcumin; A, ascorbic acid; Q, quercetin.

The cyto-protective effects of LH on ovarian reserve and female fertility during exposure to gonadotoxic alkylating agents in an adult mouse model.

STUDY QUESTION: Does LH protect mouse oocytes and female fertility from alkylating chemotherapy? SUMMARY ANSWER: LH treatment before and during chemotherapy prevents detrimental effects on follicles and reproductive lifespan. WHAT IS KNOWN ALREADY: Chemotherapies can damage the ovary, resulting in premature ovarian failure and reduced fertility in cancer survivors. LH was recently suggested to protect prepubertal mouse follicles from chemotoxic effects of cisplatin treatment. STUDY DESIGN, SIZE, DURATION: This experimental study investigated LH effects on primordial follicles exposed to chemotherapy. Seven-week-old CD-1 female mice were randomly allocated to four experimental groups: Control (n = 13), chemotherapy (ChT, n = 15), ChT+LH-1x (n = 15), and ChT+LH-5x (n = 8). To induce primary ovarian insufficiency (POI), animals in the ChT and ChT+LH groups were intraperitoneally injected with 120 mg/kg of cyclophosphamide and 12 mg/kg of busulfan, while control mice received vehicle. For LH treatment, the ChT+LH-1x and ChT+LH-5x animals received a 1 or 5 IU LH dose, respectively, before chemotherapy, then a second LH injection administered with chemotherapy 24 h later. Then, two animals/group were euthanized at 12 and 24 h to investigate the early ovarian response to LH, while remaining mice were housed for 30 days to evaluate short- and long-term reproductive outcomes. The effects of LH and chemotherapy on growing-stage follicles were analyzed in a parallel experiment. Seven-week-old NOD-SCID female mice were allocated to control (n = 5), ChT (n = 5), and ChT+LH-1x (n = 6) groups. Animals were treated as described above, but maintained for 7 days before reproductive assessment. PARTICIPANTS/MATERIALS, SETTING, METHODS: In the first experiment, follicular damage (phosphorylated H2AX histone (γH2AX) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay), apoptotic biomarkers (western blot), and DNA repair pathways (western blot and RT-qPCR) were assessed in ovaries collected at 12 and 24 h to determine early ovarian responses to LH. Thirty days after treatments, remaining mice were stimulated (10 IU of pregnant mare serum gonadotropin (PMSG) and 10 IU of hCG) and mated to collect ovaries, oocytes, and embryos. Histological analysis was performed on ovarian samples to investigate follicular populations and stromal status, and meiotic spindle and chromosome alignment was measured in oocytes by confocal microscopy. Long-term effects were monitored by assessing pregnancy rate and litter size during six consecutive breeding attempts. In the second experiment, mice were stimulated and mated 7 days after treatments and ovaries, oocytes, and embryos were collected. Follicular numbers, follicular protection (DNA damage and apoptosis by H2AX staining and TUNEL assay, respectively), and ovarian stroma were assessed. Oocyte quality was determined by confocal analysis. MAIN RESULTS AND THE ROLE OF CHANCE: LH treatment was sufficient to preserve ovarian reserve and follicular development, avoid atresia, and restore ovulation and meiotic spindle configuration in mature oocytes exposed at the primordial stage. LH improved the cumulative pregnancy rate and litter size in six consecutive breeding rounds, confirming the potential of LH treatment to preserve fertility. This protective effect appeared to be mediated by an enhanced early DNA repair response, via homologous recombination, and generation of anti-apoptotic signals in the ovary a few hours after injury with chemotherapy. This response ameliorated the chemotherapy-induced increase in DNA-damaged oocytes and apoptotic granulosa cells. LH treatment also protected growing follicles from chemotherapy. LH reversed the chemotherapy-induced depletion of primordial and primary follicular subpopulations, reduced oocyte DNA damage and granulosa cell apoptosis, restored mature oocyte cohort size, and improved meiotic spindle properties. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: This was a preliminary study performed with mouse ovarian samples. Therefore, preclinical research with human samples is required for validation. WIDER IMPLICATIONS OF THE FINDINGS: The current study tested if LH could protect the adult mouse ovarian reserve and reproductive lifespan from alkylating chemotherapy. These findings highlight the therapeutic potential of LH as a complementary non-surgical strategy for preserving fertility in female cancer patients. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by grants from the Regional Valencian Ministry of Education (PROMETEO/2018/137), the Spanish Ministry of Science and Innovation (CP19/00141), and the Spanish Ministry of Education, Culture and Sports (FPU16/05264). The authors declare no conflict of interest.

Difficult intravenous access in a paediatric intensive care unit. / Vía venosa difícil en una unidad de cuidados intensivos pediátricos.

INTRODUCTION: Multiple attempts during peripheral cannulation can have major consequences for patients, relatives, and healthcare professionals, therefore we set out to determine the extent of this problem in a paediatric intensive care unit (PICU). OBJECTIVES: The main aim was to describe peripheral venous catheter (PVC) and peripherally inserted central catheter (PICC) cannulation in children in the PICU. Secondary objectives were to determine the success rate of the first cannulation attempt, to quantify patients with difficult venous access (DVA), and to explore the association between DVA and sociodemographic, technique and nursing-related characteristics. METHOD: A cross-sectional descriptive study. Consecutive sampling was used to recruit patients aged 0-18 years admitted to the PICU who required peripheral venous cannulation. An ad hoc questionnaire was used for this purpose, including the presence of DVA as an independent variable. RESULTS: A total of 163 venous cannulations were reported. A total of 55.8% (91) were performed in patients under 1 year of age. Of these, 38.7% (63) were successful on the first attempt and 36.8% (60) had DVA. When there was DVA, 85% (51) of patients had complications, median time to cannulation by short CVP was 30minutes [15-53] and 2 or more nurses were required on 80% (48) of occasions. CONCLUSIONS: We found a low success rate at first attempt and a high proportion of DVA. More nurses and time were employed during cannulation and complications increased if the patient had DVA. A statistically significant association was found between DVA and age, weight, poor perfusion, veins that were neither visible nor palpable, DIVA score≥4, history of difficult intravenous access, complications, number of nurses and time spent.

Differential effects of the immunosuppressive calcineurin inhibitors cyclosporine-A and tacrolimus on ovulation in a murine model.

STUDY QUESTION: Do therapeutic levels of cyclosporine-A and tacrolimus affect ovulation in a rat gonadotrophin-induced ovulation model? SUMMARY ANSWER: Cyclosporine-A, but not tacrolimus, decreases ovulation rate when administered for 5 days before induced ovulation. WHAT IS KNOWN ALREADY: The mainstays of immunosuppression in solid organ transplantation, to prevent rejection, are the calcineurin inhibitors cyclosporine-A or tacrolimus. These drugs could potentially affect fertility in transplanted patients. Since ovulation is an inflammation-like process with pivotal roles for several immune cells and modulators, it is possible that the calcineurin inhibitors, with broad effects on the immune system, could interfere with this sensitive, biological process. STUDY DESIGN SIZE DURATION: Experimental design at university-based animal facilities. A total of 45 immature Sprague-Dawley rats were used. The study was carried out over 3 months. PARTICIPANTS/MATERIALS SETTING METHODS: Immature Sprague-Dawley rats (n = 45) were randomly assigned to receive equivalent doses of tacrolimus (0.5 mg/kg/day; TAC), cyclosporine-A (10 mg/kg/day; CyA) or vehicle (Control). Ovarian hyperstimulation was induced with 10 IU of equine chorionic gonadotrophin, and ovulation was triggered with 10 IU of hCG. Oocytes were retrieved from the oviducts and ovulation rates were calculated. Various subpopulations of white blood cells were counted in peripheral blood and ovarian tissue samples. MAIN RESULTS AND THE ROLE OF CHANCE: Animals in the CyA group showed a lower ovulation rate when compared to the TAC and Control groups (CyA: mean 9 oocytes (range 0-22); TAC: 21 oocytes (8-41); Control: 22 oocytes (6-39); P = 0.03). Regarding counts of the white blood cell subpopulations and resident neutrophils in the ovary, no significant differences were observed between the groups. LIMITATIONS REASONS FOR CAUTION: Although the ovulation process is highly conserved within species, the differences between rodents and humans may limit the external translatability of the study. WIDER IMPLICATIONS OF THE FINDINGS: These findings suggest that tacrolimus should be the preferred calcineurin inhibitor of choice in transplanted patients who are aiming for pregnancy. STUDY FUNDING/COMPETING INTERESTS: Swedish Research Council and ALF of Sahlgrenska Academy, Sweden. Rio Hortega Grant from the Instituto de Salud Carlos III, Spain (CM09/00063). There are no conflicts of interest.

Passivated graphene quantum dots for carbaryl determination in juices.

This paper reports a simple method for the preparation of suitable graphene quantum dots after surface passivation, to be used for the determination of carbaryl in juice samples. A comparison of synthetic conditions for the preparation of graphene quantum dots following the top-down approach is described. In the one-step route selected, evaluation of diverse reaction time for cutting and modulating the oxidizing sites in the broken pieces of the initial graphene layer is conducted with a mixture of concentrated acids. Exploring the passivation effect on the purified graphene quantum dots, we demonstrated the suitability of the selected graphene quantum dots for practical application in the detection of carbaryl using fluorometric detection. Higher sensitivity was achieved after 8 min of contact, in which graphene quantum dots promotes the degradation of carbaryl into naphthol, being the latter responsible for the analytical signal. The detection and quantification limits were 0.36 and 1.21 µg/L, respectively, being the response linear up to 26 µg/L with excellent precision (better than 3.2% at the limit of detection). The recovery of the analyte from commercial juice samples (91.4-96.7%) testifies to the applicability of the proposal for the analytical problem selected.

Impact of patients' perception of COPD and treatment on adherence and health-related quality of life in real-world: Study in 53 community pharmacies.

BACKGROUND: Patient adherence to treatment in chronic obstructive pulmonary disease (COPD) is essential to optimize disease management. We aimed to assess the impact of patients' perception of their treatment and disease on adherence and Health-Related Quality of Life (HRQL) in patients attending a community pharmacy, where usually subjects have a better condition than those in clinical settings. METHODS: We performed a cross-sectional study of 318 patients with COPD in treatment with inhalers in the last 3 months from 53 community pharmacies. We assessed HRQL with St George's Respiratory Questionnaire (SGRQ). Persistence was assessed from the three previous refills and adherence through the Test of Adherence to Inhalers test. RESULTS: Persistence was achieved by 78.6% of the patients and 58.5% had good adherence. Patients having a multidose DPI and those with MDI showed a 2.8-fold and 4.1-fold increased association, respectively, with intermediate/poor adherence in comparison with those having a single dose DPI. Those patients who did not have knowledge about COPD (aOR 2.106, p = 0.006) and those who thought that the inhaler effectiveness was fair/poor (aOR 2.361, p = 0.006) were more likely to have intermediate/poor adherence. Overall SGRQ score was significantly worse in patients with intermediate/poor adherence (p = 0.036) and in those who thought the inhaler's effectiveness was fair/poor (p < 0.001). CONCLUSIONS: The type of inhaler and patients' knowledge and perceptions of their disease and treatment were associated with good adherence and higher HRQL. Clinicians should promote shared-decision making in the choice of inhaler depending on patients' individual abilities and beliefs.

Handwritten Geometrical Patterns in the Evaluation of Motor Symptoms in Psychotic Disorders.

The analysis of handwriting has been used in several contexts. For example, handwriting has shown to be of value in the study of motor symptoms in neurological and mental disorders. In the present work, the geometric analysis of handwriting patterns is proposed as a tool to evaluate motor symptoms in psychotic disorders. Specifically, we have employed the lacunarity, a measure of the heterogeneity of a spatial structure. Forty-two patients with a psychotic disorder and 35 matched healthy controls participated in the study. Participants were asked to copy some patterns with a pen on a white paper. The results showed that lacunarity was significantly higher in handwritten patterns from patients than from controls. In addition, we found higher values of lacunarity in handwritten patterns from patients with severe motor symptoms in comparison with patients with mild or absent motor symptoms. Lacunarity of handwritten patterns was significantly correlated with clinical scores of rigidity. In conclusion we argue that the heterogeneity of handwritten patterns could be used as a simple and objective measure of motor symptoms.

Cotton fibers functionalized with ß-cyclodextrins as selectivity enhancer for the direct infusion mass spectrometric determination of cocaine and methamphetamine in saliva samples.

Cotton is a natural material with good mechanical and chemical properties. It presents a hydrophilic surface that must be modified by the introduction of appropriate groups, if semi-polar or non-polar analytes are intended to be isolated from an aqueous matrix. In this article, cotton is modified with ß-cyclodextrins, which presents a hydrophobic cavity where target molecules can be hosted. The cavity size restricts the access of larger or less accessible molecules due to their conformation, thus improving the extraction selectivity. The modified cotton fibers are packed in a disposable syringe device where all the steps of the extraction take place. The ensuing fibers were evaluated for the extraction of several drugs providing the best results for cocaine (CO) and methamphetamine (MTA). These targets were extracted from saliva samples and finally determined by direct infusion mass spectrometry. The method provides limits of detection and quantification of 0.6 and 2 µg L-1 for CO and 0.8 and 2.7 µg L-1 for MTA, with precision values at the quantification level better than the 9% (expressed as relative standard deviation). Also, relative recoveries ranged from 115% to 111% for CO and MTA, respectively, demonstrating the applicability of the proposed method. Matrix effect, which has been statistically evaluated, is not significant, showing that the extraction selectivity can compensate the circumvent of the chromatographic separation.

Discrimination between nanocurcumin and free curcumin using graphene quantum dots as a selective fluorescence probe.

Accurate-controlled sized graphene quantum dots (GQDs) have been used as an analytical nanoprobe for detecting curcumin as a function of the photoluminescent quenching upon increasing concentrations of the analyte. Regarding the importance of curcumin nanoparticles in nutraceutical food, the analytical method described herein was also proven for the discrimination of curcumin remaining in free solution from that encapsulated into water-soluble nanomicelles of ca. 11 nm. This recognition is based on the displacement of GQD emission when interacting with both curcumin species. Maximum emission wavelength of GQDs suffers a gradual quenching as well as a red-shifting upon increasing concentrations of free curcumin (from 458 to 490 nm, exciting at 356 nm). On the other hand, in the presence of nanocurcumin, GQD photoluminescent response only displays a quenching effect (458/356 nm). The sensitivity of the described method in terms of detection limits was 0.3 and 0.1 µg mL-1 for curcumin and nanocurcumin, respectively. The applicability of the photoluminescent probe for the quantification and discrimination between both curcumin environments was demonstrated in nutraceutical formulations namely functional food capsules and fortified beverages such as ginger tea. Graphical abstract.

Effect of biocrusts on bacterial community composition at different soil depths in Mediterranean semi-arid ecosystems.

This study analyzed the influence of biocrusts on the chemical properties and bacterial diversity and community composition in the underlying soils along a depth gradient (the biocrust (C1), middle (S2) and deep (S3) soil layers) in two semi-arid Mediterranean ecosystems. Organic carbon, pH, electric conductivity and calcium carbonate content were estimated by wet oxidation, potentiometrically (pHmeter), with a conductivity-meter and volumetrically with a Bernard calcimeter, respectively. Bacterial diversity and community composition were estimated by 16S rRNA gene high-throughput amplicon sequencing. Chemical properties in C1 were significantly different from the other soil layers, showing higher organic carbon content and lower pH (p < 0.05). The relative abundance of several bacterial taxa, such as Bryocella, Methylobacterium, Segitebacter and Actinomycetospora showed significant positive correlations with organic carbon (r = 0.53 to 0.75) and negative with pH (r = -0.72 to -0.84), and were also highly correlated with each other (p < 0.01), suggesting a bacterial co-occurrence pattern associated with the biocrust. On the contrary, other bacterial taxa, such as Euzebyaceae, Truepera, Alphaproteobacteria and Caldinilaceae, showed positive correlations with electrical conductivity and calcium carbonate and were also correlated with each other (p < 0.01), in a second type of co-occurrence pattern associated with bare soil. The C1 and S2 layers had several taxa in common, while S3 layers had taxa common to bare soil, suggesting that the effect of biocrusts was limited to the first centimeters of soil and progressively decreased in depth. Bacterial diversity was lower in C1 than in the underlying layers and increased progressively from biocrust to deeper soil layers. The results suggest that the diversity and composition of soil microbial communities in biologically crusted sites in Mediterranean semi-arid environments are mainly controlled by chemical properties which in turn are modified by the biocrust along a depth gradient.