When residents die in Portuguese care facilities: The experiences and practices of care facility directors before and during the COVID-19 pandemic.

This qualitative exploratory study aimed to describe the experiences and practices of care facility directors regarding residents dying in Portuguese residential care facilities (RCFs) before and during the COVID-19 pandemic. Data was obtained from 17 care director facilities who participated in focus groups. The data analysis followed the interpretative phenomenological approach. Participants stated that practices surrounding the death of the residents in Portuguese RCFs changed significantly from before to during the pandemic, regarding both those who died from COVID-19 infection and from other conditions. Four themes emerged that illustrated the situation during the peaks of the pandemic. During the pandemic, practices and experiences quite different from the usual ones have emerged. These results support the importance of a good death, and dying with dignity in RCFs, and the need for policies, practices, and training on the death of residents in RCFs in Portugal.

Cellulose-based hydrogel on quantum dots with molecularly imprinted polymers for the detection of CA19-9 protein cancer biomarker.

Molecularly imprinted polymers MIPs were successfully assembled around quantum dots (QDs), for the detection of the protein biomarker CA19-9 associated to pancreatic cancer (PC). These imprinted materials MIP@QDs were incorporated within the cellulose hydrogel with retention of its conformational structure inside the binding cavities. The concept is to use MIPs which function as the biorecognition elements, conjugated to cadmium telluride QDs as the sensing system. The excitation wavelength was set to 477 nm and the fluorescence signal was measured at its maximum intensity, with an emission range between 530 and 780 nm. The fluorescence quenching of the imprinted cellulose hydrogels occurred with increasing concentrations of CA19-9, showing linearity in the range 2.76 × 10 -2 - 5.23 × 10 2 U/ml, in a 1000-fold diluted human serum. Replicates of the imprinted hydrogel show a linear response below the cut-off values for pancreatic cancer diagnosis (< 23 U/ml), a limit of detection of 1.58 × 10 -3 U/ml and an imprinting factor (IF) of 1.76. In addition to the fact that the imprinted cellulose hydrogel displays good stability and selectivity towards CA19-9 when compared with the non-imprinted controls, the conjugation of MIPs to QDs increases the sensitivity of the system for an optical detection method towards ranges within clinical significance. This fact shows potential for the imprinted hydrogel to be applied as a sensitive, low-cost format for point-of-care tests (PoCTs).

Assessment of water quality in Aguieira reservoir: Ecotoxicological tools in addition to the Water Framework Directive.

To assess the ecological potential (EP), the Water Framework Directive (WFD) uses specific parameters but ignores ecosystem functioning and individual/subindividual biological responses. The heavily modified water bodies (reservoirs) are strongly influenced by human activities that promote their imbalance, with a loss or compromise the biodiversity and the functioning of these ecosystems. This work aims to determinate the EP according physical, chemical and biological parameters of WFD and evaluate the functionality and sensitivity of ecotoxicological tools in the evaluation of water quality of Aguieira reservoir. Four sampling sites were defined in Aguieira reservoir and sampling was conducted in autumn 2018, spring and autumn 2019. WFD showed overall that Aguieira reservoir presented moderate EP. The autumn samples and the two reservoir upstream sites had the worst classification. These results were complemented by a set of ecotoxicological assays (growth inhibition of Raphidocelis subcapitata and Spirodela polyrhiza, and feeding rate of Daphnia longispina) using three natural water treatments. R. subcapitata was the most sensitive species, indicating the presence of a risk that is not detected by chemical analysis, nor by organisms of different trophic levels. D. longispina only showed sensitivity in A3 and A4 in autumn, however this results was difficult to interpret since no reason can be observed to respond the F1 lower feeding rate observed in almost sites. S. polyrhiza, as well as the total chlorophyll, lipid peroxidation, and proline content did not show sensitivity, since no discrimination among the water treatments were recorded. The relevance of bioassays is associated with the fact that an integrated response to a complex mixture can be obtained, which can contain more substances than the measured ones. Thus, we may conclude that WFD are insufficient to draw conclusions about water quality, and the growth inhibition of Raphidocelis subcapitata assay is an important parameter to complement the WFD. In addition, species of different trophic levels are recommended for ecotoxicological analyses due to differences in species sensitivities.

Can biochemical endpoints improve the sensitivity of the biomonitoring strategy using bioassays with standard species, for water quality evaluation?

The Water Framework Directive (WFD) was adopted in 2000 and is a common framework for water policy, management and protection in Europe. The WFD assesses specific parameters; however, it ignores indicators of ecosystem functioning and sub-individual performance. Reservoirs are strongly influenced by anthropogenic activities that promote their imbalance. Bioassays and biomarkers are useful tools to link the chemical, ecological and toxicological assessments in water quality assessments. These approaches can be complementary to WFD methodologies, allowing the detection of impacts on the ecosystem. This study evaluated if the biochemical parameters can improve the sensitivity of the biomonitoring strategy using bioassays with the standard species Daphnia magna, in the assessment of the ecological quality of water reservoirs. To this end, water samples of Portuguese reservoirs were analysed in three sampling periods (Autumn 2018 and Spring, Autumn 2019). In parallel, a physicochemical characterization of waters was performed. D. magna feeding rate assays were performed for 24 h. After exposure, metabolism, oxidative stress and lipid peroxidation biomarkers were evaluated. Feeding rate assays showed sensitivity to different reservoirs. Biomarkers showed a higher sensitivity and can therefore improve the sensitivity of the biomonitoring strategy using bioassays. Bioassays and biomarkers approach allowed to highlight potential sources of stress, more related to the quality of the seston than to chemical contamination. This work highlights the complementarity between bioassays and biomarkers to identify ecotoxicological effects of surface waters, and can be extremely useful, especially in cases where the biotic indices are difficult to establish, such as reservoirs.

Distinctive features of positive anti-cell antibody tests (indirect immunofluorescence on HEp-2 cells) in patients with non-autoimmune diseases.

OBJECTIVE: The objective of this study was to determine whether characteristics of positive results in the indirect immunofluorescence assay on HEp-2 cells for anti-cell antibodies (HEp-2 IFA) differ between patients with non-autoimmune diseases (NADs) and patients with systemic autoimmune rheumatic diseases (SARDs). METHODS: Cross-sectional observational study comparing HEp-2 IFA test results in three groups: (a) 558 NAD patients comprising four subgroups (cancer ( n = 95), infectious diseases ( n = 148), psychiatric diseases ( n = 163), common non-infectious chronic diseases ( n = 152)); (b) 194 SARD patients; (c) 1217 healthy individuals (HIs). Sera were tested at 1:80 dilution and diluted to the end titer. Slides were analyzed by two independent blinded examiners. RESULTS: A positive HEp-2 IFA test occurred in 102 (18.3%) NAD patients, 170 (87.6%) SARD patients and 150 (12.3%) HIs. The four NAD subgroups did not differ regarding HEp-2 IFA frequency, titer or pattern. HEp-2 IFA titer was higher in NAD patients than in HIs and both had lower titer than SARD patients. Nuclear dense fine speckled pattern was more frequent in NAD patients and HIs than in SARD patients ( p < 0.001). Nuclear homogeneous and nuclear coarse speckled patterns were more frequent in SARD patients than in the other groups ( p < 0.001). The nuclear fine speckled pattern was prevalent in all three groups, but presented a gradient in titer across them; HIs and NAD patients had low and intermediary titers, which were significantly lower than in SARD patients ( p < 0.001). CONCLUSION: Positive HEp-2 IFA frequency, pattern and titer present differential features in NAD and SARD patients, and this attribute adds value to the test in the diagnosis of SARDs.

Histone acetylation increases in response to ferulic, gallic, and sinapic acids acting synergistically in vitro to inhibit Candida albicans yeast-to-hyphae transition.

Novel treatments are needed to prevent candidiasis/candidemia infection due to the emergence of Candida species resistant to current antifungals. Considering the yeast-to-hyphae switch is a critical factor to Candida albicans virulence, phenols common in plant sources have been reported to demonstrating their ability to prevent dimorphism. Therefore, phenols present in many agricultural waste stress (ferulic (FA) and gallic (GA) acid) were initially screened in isolation for their yeast-to-hyphae inhibitory properties at times 3, 6, and 24 hr. Both FA and GA inhibited 50% of hyphae formation inhibitory concentration (IC50 ) but at a concentration of 8.0 ± 0.09 and 90.6 ± 1.05 mM, respectively, at 24 hr. However, the inhibitory effect of FA increased by 1.9-2.6 fold when combined with different GA concentrations. GA and FA values decreased even lower when sinapic acid (SA) was added as a third component. As evidenced by concave isobolograms and combination indexes less than 1, both GA:F A and GA:FA:SA combinations acted synergistically to inhibit 50% hyphae formation at 24 hr. Lastly, acetylation of histone H3 lysine 56 acetylation (H3K56) was higher in response to the triple phenolic cocktail (using the IC50 24 hr inhibitory concentration level) comparable with the nontreated samples, indicating that the phenols inhibited hyphal growth in part by targeting H3K56 acetylation.

Untangling Neolithic and Bronze Age mitochondrial lineages in South Asia.

Two key moments shaped the extant South Asian gene pool within the last 10 thousand years (ka): the Neolithic period, with the advent of agriculture and the rise of the Harappan/Indus Valley Civilisation; and Late Bronze Age events that witnessed the abrupt fall of the Harappan Civilisation and the arrival of Indo-European speakers. This study focuses on the phylogeographic patterns of mitochondrial haplogroups H2 and H13 in the Indian Subcontinent and incorporates evidence from recently released ancient genomes from Central and South Asia. It found signals of Neolithic arrivals from Iran and later movements in the Bronze Age from Central Asia that derived ultimately from the Steppe. This study shows how a detailed mtDNA phylogeographic approach, combining both modern and ancient variation, can provide evidence of population movements, even in a scenario of strong male bias such as in the case of the Bronze Age Steppe dispersals.

Biochemical alterations of Candida albicans during the phenotypic transition from yeast to hyphae captured by Fourier transform mid-infrared-attenuated reflectance spectroscopy.

Candida albicans is an opportunistic human pathogen that can become virulent due to its ability to switch from a benign yeast to virulent hyphae phenotype. The emergence of C. albicans resistant to commonly used antifungal agents has necessitated the development of innovative treatments, which must be accompanied by an understanding of the molecular changes that occur during the phenotypic shift. For this purpose, Fourier transform mid-infrared spectroscopy in attenuated total reflectance mode (FT-mIR-ATR) was applied to monitor the structural and compositional changes in C. albicans during the yeast-to-hyphae transition. FT-mIR-ATR measurements were completed on the whole cell of C. albicans (SC5314) during hyphal formation induced by N-acetylglucosamine at 0, 1, 2, 3, 6 and 24 h. Principal component analysis separated the FT-mIR-ATR spectra into four groups that were aligned with the morphological changes captured by microscopic imaging. Spectral signatures indicating the structural and compositional modifications during the transition were identified mainly in the fatty acid region (3100-2800 cm-1), the protein and peptide region (1800-1500 cm-1), the mixed region (1500-1200 cm-1) and the polysaccharide region (1200-900 cm-1). A spectral fingerprint of the transition via a heat map was generated based on the peak shift in position. Quantitative evaluation of the spectra by curve fitting further revealed the dynamics of the cell's main components during the transition. This work provides valuable structural and functional information on the C. albicans phenotypic transition to hyphae, which has diagnostic implications.

In vivo Confocal Laser Microscopy for monitoring of actinic keratosis treatment: a comparison with histopathologic assessment after treatment with topical 5% 5-fluorouracil.

BACKGROUND: Histological examination is the gold standard for actinic keratosis diagnosis; however, it is not always a feasible approach. Reflectance confocal microscopy (RCM) is a non-invasive technique that may be an alternative for monitoring actinic keratoses treatment response. Topical 5-fluorouracil is indicated for actinic keratosis multiple lesions and for field cancerization treatment. OBJECTIVES: To assess the RCM accuracy, sensibility and specificity for actinic keratosis, considering as a gold standard the histopathological examination; as well as to evaluate the efficacy of 5% 5-fluorouracil treatment. METHODS: This is a prospective study in actinic keratosis patients between August 2014 and November 2015. RCM analyses were performed in one randomly selected actinic keratosis lesion of the upper limbs by two independent observers before and after 5% 5-fluorouracil treatment. At the end of treatment and with clinical bleaching of treated lesions, histological examination was performed by two pathologists. RESULTS: A total of 50 lesions were enroled, and 40 lesions presented complete clinical bleaching after treatment and were included in the final analysis. Accuracy, sensibility and specificity means among observers were 83.8%, 84.6% and 83.3%, respectively. After 5-fluorouracil treatment, actinic keratosis was diagnosed in 45.0% (observer 1) and 32.5% (observer 2) of subjects according to RCM and in 32.5% of subjects according to histological examination. Considering RCM observers diagnosis, the concordance was substantial (k 0.637, P < 0.001). 5-fluorouracil led to a reduction in 55.0%-67.5% of actinic keratoses according to RCM analysis. CONCLUSION: This study allows to validate RCM as a non-invasive method capable of monitoring actinic keratosis therapeutic response to 5-fluorouracil, presenting efficacy comparable to histological examination. Additionally, the results suggest that 5-fluorouracil may be a satisfactory option for therapeutic control of this condition.

Reducing prepartum urine pH by supplementing anionic feed ingredients: Effects on physiological and productive responses of Holstein × Gir cows.

This study compared physiological and productive parameters in 3/4 Holstein × 1/4 Gir dairy cows receiving a prepartum concentrate containing ammonium chloride to reduce urine pH near 7.0 (CON; n = 17), or a commercial anionic supplement to reduce urine pH near 6.0 (SUPP; n = 17). Nonlactating, multiparous, pregnant cows were assigned to receive SUPP or CON beginning 21 d before expected date of calving. Cows were maintained in a single drylot pen with ad libitum access to corn silage, and individually received their prepartum concentrate once daily (0800 h) before calving. Cows from both treatments completely consumed their concentrate allocation within 30 min after feeding. Cow body weight and body condition score were recorded once weekly, urine pH measured every 3 d, and blood samples collected on d -21, -14, -9, -6, and -3 relative to expected calving date. After calving (d 0), cows were moved to an adjacent drylot pen with ad libitum access to water and a total mixed ration, and were milked twice daily (0600 and 1700 h). Cow body weight and body condition score were recorded once weekly and individual milk production was recorded daily until 30 d in milk (DIM). Blood samples were collected before each milking during the first 5 DIM, as well as at 6, 9, 16, 23, and 30 DIM before the morning milking. Based on actual calving dates, cows received SUPP or CON for (mean ± standard error) 19.2 ± 1.2 and 19.0 ± 0.9 d before calving, respectively. Urine pH was less in SUPP versus CON cows during the last 15 d of gestation (6.12 vs. 7.15, respectively). Milk yield during the first 5 DIM and throughout the experimental period was greater in SUPP versus CON cows (by 20 and 14%, respectively), whereas serum Ca concentrations did not differ between treatments during the first 5 DIM. Serum concentrations of fatty acids were greater in SUPP versus CON cows 3 d before and at calving (by 52 and 22%, respectively), whereas SUPP cows had lower serum glucose and cortisol concentration at calving (by 23 and 27%, respectively). Hence, the SUPP treatment decreased prepartum urine pH near 6.0 in Holstein × Gir dairy cows without depressing concentrate intake compared with CON, although total dry matter intake was not evaluated to fully investigate feed intake responses. Moreover, the SUPP treatment transiently affected serum glucose, fatty acids, and cortisol concentrations near the time of calving, and resulted in greater milk yield during the initial 30 DIM compared with CON.

Effects of supplemental calcium salts of palm oil and chromium-propionate on insulin sensitivity and productive and reproductive traits of mid- to late-lactating Holstein × Gir dairy cows consuming excessive energy.

This experiment compared insulin sensitivity, milk production, and reproductive outcomes in dairy cows consuming excessive energy during mid to late lactation and receiving in a 2 × 2 factorial design (1) concentrate based on ground corn (CRN; n = 20) or including 8% (DM basis) of Ca salts of palm oil (CSPO; n = 20), and (2) supplemented (n = 20) or not (n = 20) with 2.5 g/d of Cr-propionate. During the experiment (d 0-203), 40 multiparous, nonpregnant, lactating 3/4 Holstein × 1/4 Gir cows (initial days in milk = 81 ± 2; mean ± SE) were offered corn silage for ad libitum consumption, and individually received concentrate formulated to allow diets to provide 160% of their daily net energy for lactation requirements. From d -15 to 203, milk production was recorded daily, blood samples collected weekly, and cow body weight (BW) and body condition score (BCS) recorded on d 0 and 203. For dry matter intake evaluation, cows from both treatments were randomly divided in 5 groups of 8 cows each, and allocated to 8 individual feeding stations for 3 d. Intake was evaluated 6 times/group. Glucose tolerance tests (GTT; 0.5 g of glucose/kg of BW) were performed on d -3, 100, and 200. Follicle aspiration for in vitro embryo production was performed via transvaginal ovum pick-up on d -1, 98, and 198. Mean DMI, net energy for lactation intake, as well as BW and BCS change were similar across treatments. On average, cows gained 40 kg of BW and 0.49 BCS during the experiment. Within weekly blood samples, CRN cows had lower serum concentrations of glucose, insulin, fatty acids, and insulin-to-glucose ratio compared with CSPO cows, suggesting increased insulin sensitivity in CRN cows. During the GTT, insulin-sensitivity traits were also greater in CRN versus CSPO cows. Supplemental Cr-propionate resulted in lower serum insulin concentrations and insulin-to-glucose ratio within CRN cows only, indicating that Cr-propionate improved basal insulin sensitivity in CRN but not in CSPO cows. During the GTT, however, Cr-propionate supplementation reduced hyperinsulinemia and insulin-to-glucose ratio across CSPO and CRN cows. Milk production, as well as number of viable oocytes collected and embryos produced within each aspiration, were not affected by treatments. Hence, replacing corn by Ca salts of palm oil in the concentrate did not improve insulin sensitivity in Holstein × Gir dairy cows consuming excessive energy during mid to late lactation, whereas Cr-supplementation was effective in improving basal insulin sensitivity in cows not receiving Ca salts of palm oil.

Oxytetracycline effects in specific biochemical pathways of detoxification, neurotransmission and energy production in Oncorhynchus mykiss.

Oxytetracycline (OTC) is a tetracycline antibiotic, widely used in human and veterinary medicines, including in aquaculture. Given this use, OTC has been detected in different aquatic environments. Some recent works have demonstrated unintentional biological activity of OTC in non-target aquatic organisms. This study investigated the acute and chronic effects of OTC on the physiology of the fish species Oncorhynchus mykiss (rainbow trout), namely through the quantification of the activity of enzymes involved in different biochemical pathways, such as detoxification (phase II - glutathione S-transferases - GSTs, uridine-diphosphate-glucuronosyltransferases - UGTs), neurotransmission (acetylcholinesterase - AChE) and energy production (lactate dehydrogenase - LDH). The here-obtained data demonstrated the induction of GSTs activity in gills, and inhibition of AChE activity in eyes tissue, in chronically exposed organisms, as well as alterations in LDH activity following both exposures. Considering this set of results, we can infer that OTC exposure may have induced the glutathione pathway of detoxification in gills with the involvement of GSTs, or indirectly due to the metabolites that may have been produced. In turn, these metabolites may have interfered with the mechanism of neurotransmission, also causing physiological and biochemical disturbances in rainbow trout after OTC exposure, namely disturbances in energetic metabolism. In addition, it is important to stress that such occurrences took place at low, environmentally realistic levels of OTC, suggesting that organisms exposed in the wild may be putative targets of toxic effects by commonly used drugs such as antibiotics.

Assessment of ecotoxicological effects of ciprofloxacin in Daphnia magna: life-history traits, biochemical and genotoxic effects.

Antibiotics (e.g. ciprofloxacin) have been detected in surface water and groundwater for several decades. In order to understand the potential impact of the continuous exposure of aquatic organisms to ciprofloxacin, a chronic assay was carried out with Daphnia magna. This approach allowed evaluation of the effects of ciprofloxacin on life-history and sub-individual parameters (antioxidant status and metabolic response: activities of catalase and glutathione S-transferases - GSTs; peroxidative damage; thiobarbituric acid reactive substances and genotoxic effects (genetic damage index, measured by the comet assay). Life-history parameters of D. magna showed no significant effects after ciprofloxacin exposure. Concerning oxidative stress and metabolism parameters, no significant alterations were reported for catalase and GSTs activities. However, a dual response was observed, with a significant decrease in lipid peroxidation levels at low ciprofloxacin concentrations (<0.013 mg/L), while a significant increase was verified at high ciprofloxacin concentrations (0.078 mg/L). The genotoxicity assay detected a significant increase in genetic damage index up to 0.013 mg/L of ciprofloxacin. The here-tested ciprofloxacin concentrations, which are ecologically relevant, did not cause significant impacts concerning the life-history parameters of D. magna; however, at the same levels of ciprofloxacin an oxidative stress and genotoxic damage scenarios were recorded.

Effect of ageing on the calibration of ballistic gelatin.

PURPOSE: Ballistic gelatin is commonly used as a validated surrogate for soft tissue during terminal ballistic testing. However, the effect of a delay between production and testing of a gelatin mould remains unknown. The aim of this study was to determine any potential effects of ageing on ballistic gelatin. METHODS: Depth of penetration (DoP) of 4.5 mm spherical fragment simulating projectiles was ascertained using mixtures of 10%, 11.25% and 20% Type A 250 Bloom ballistic gelatin. Testing was performed daily for 5 days using velocities between 75 and 210 m/s. DoP at day 5 was statistically compared with day 1, and net mass change was recorded daily. RESULTS: No significant difference was found for DoP observed with time in any of the samples (P>0.05). Spearman correlation was excellent in all moulds. The moulds with known standard calibrations remained in calibration throughout the study period. Mass loss of less than 1% was noted in all samples. CONCLUSION: Mass loss was the only quantifiable measure of changes in the blocks with time, but did not correlate with any changes in DoP. This may provide reassurance when undertaking such testing that an inadvertent delay will not significantly alter the penetration properties of the mould. Future research is recommended to determine any potential effect on the mechanical properties of gelatin at higher velocity impacts and whether the calibration corresponds to an adequate simulation under such conditions.

Liver transplant recipients have a higher prevalence of anal squamous intraepithelial lesions.

BACKGROUND: Anal squamous intraepithelial lesions (ASIL) are precancerous lesions of anal squamous cell carcinoma, with a higher prevalence in immunosuppressed patients. There are some studies in kidney transplant recipients, but there is no information regarding prevalence in liver transplantation. Our aim was to evaluate the prevalence of ASIL in this setting. METHODS: Prospective case-control study involving liver transplant recipients without any other known risk factor for ASIL (n=59), which were compared with a healthy control group (n=57). All were submitted to anal cytology and high-resolution anoscopy was performed in those with abnormal results. RESULTS: Ten (17%) of liver transplant recipients had abnormal cytological results, seven patients had atypical squamous cells of undetermined significance (ASC-US), one patient had atypical squamous cells that cannot exclude high-grade (ASC-H) and two patients had high-grade squamous intraepithelial lesions (HSIL). In the control group, one patient (2%) had an ASC-US result (P=0.005). Anal squamous intraepithelial lesions were confirmed in 7 out of 10 of liver transplant patients and 0 out of 1 in the controls (P=0.013) by high-resolution anoscopy with biopsies. Current smoking was the only risk factor for abnormal cytology (odds ratio=5.87, 95% confidence intervals=1.22-28.12, P=0.027). CONCLUSIONS: Liver transplant patients have a higher risk of ASIL. Screening should be considered, especially in smokers.

Protective effects of exogenous and endogenous hydrogen sulfide in mast cell-mediated pruritus and cutaneous acute inflammation in mice.

The recently described 'gasomediator' hydrogen sulfide (H2S) has been involved in pain mechanisms, but its effect on pruritus, a sensory modality that similarly to pain acts as a protective mechanism, is poorly known and controversial. The effects of the slow-releasing (GYY4137) and spontaneous H2S donors (Na2S and Lawesson's reagent, LR) were evaluated in histamine and compound 48/80 (C48/80)-dependent dorsal skin pruritus and inflammation in male BALB/c mice. Animals were intradermally (i.d.) injected with C48/80 (3µg/site) or histamine (1µmol/site) alone or co-injected with Na2S, LR or GYY4137 (within the 0.3-100nmol range). The involvement of endogenous H2S and KATP channel-dependent mechanism were also evaluated. Pruritus was assessed by the number of scratching bouts, whilst skin inflammation was evaluated by the extravascular accumulation of intravenously injected 125I-albumin (plasma extravasation) and myeloperoxidase (MPO) activity (neutrophil recruitment). Histamine or C48/80 significantly evoked itching behavior paralleled by plasma extravasation and increased MPO activity. Na2S and LR significantly ameliorated histamine or C48/80-induced pruritus and inflammation, although these effects were less pronounced or absent with GYY4137. Inhibition of endogenous H2S synthesis increased both Tyrode and C48/80-induced responses in the skin, whereas the blockade of KATP channels by glibenclamide did not. H2S-releasing donors significantly attenuate C48/80-induced mast cell degranulation either in vivo or in vitro. We provide first evidences that H2S donors confer protective effect against histamine-mediated acute pruritus and cutaneous inflammation. These effects can be mediated, at least in part, by stabilizing mast cells, known to contain multiple mediators and to be primary initiators of allergic processes, thus making of H2S donors a potential alternative/complementary therapy for treating inflammatory allergic skin diseases and related pruritus.

Laparoscopic suturing learning curve in an open versus closed box trainer.

BACKGROUND: The aim of this study was to examine the influence of training under direct vision prior to training with indirect vision on the learning curve of the laparoscopic suture task. METHODS: Novices were randomized in two groups. Group 1 performed three suturing tasks in a transparent laparoscopic box trainer under direct vision followed by three suturing tasks in a standard non-transparent laparoscopic box trainer equipped with a 0° laparoscope. Group 2 performed six suturing tasks in a standard laparoscopic box trainer. Performance time, motion analysis parameters (economy of movements) and interaction force parameters (tissue handling) were measured. Participants completed a questionnaire assessing: self-perceived dexterity before and after the training, their experienced frustration and the difficulty of the training. RESULTS: A total of 34 participants were included, one was excluded because of incomplete training. Group 1 used significantly less time to complete the total of six tasks (27 %). At the end of the training, there were no differences in motion or force parameters between the two groups. Group 2 rated their self-perceived dexterity after the training significantly lower than before the training and also reported significantly higher levels of frustration compared to group 1. Both groups rated the difficulty of the training similar. CONCLUSION: Novices benefit from starting their training of difficult basic laparoscopic skills, e.g., suturing, in a transparent box trainer without camera. It takes less time to complete the tasks, and they get less frustrated by the training with the same results on their economy of movements and tissue handling skills.

Pulpectomies in primary mandibular molars: a comparison of outcomes using three root filling materials.

AIM: To evaluate the outcome of root canal treatment in primary teeth using three root canal filling materials - RC Fill, Vitapex and Pulpdent root canal sealer. METHODOLOGY: The study was a single-centre, double-blind, randomized controlled trial carried out on 129 primary mandibular molars with necrotic pulps or irreversible pulpitis in 4- to 9-year-old children. Participants were selected based on specific inclusion and exclusion criteria and were randomly allocated into 3 groups: Group I - RC Fill [zinc oxide-eugenol (ZOE) with iodoform]; Group II - Vitapex (calcium hydroxide with iodoform); and Group III - Pulpdent root canal sealer (ZOE). The outcome measures were evaluated both clinically and radiographically at 6, 12 and 30 months according to modified American Association of Endodontists (AAE) criteria. The radiographic outcomes were assessed by two blinded and calibrated evaluators. Pearson's chi-square analysis was performed for both intention-to-treat (ITT) and per-protocol population. RESULT: The success rates of RC Fill, Vitapex and Pulpdent were 94%, 90% and 97%, respectively, at 30 months and the differences were not significant (P = 0.137). CONCLUSION: All three materials were found to be equally effective root filling materials for primary molars with necrotic pulps and irreversible pulpitis at 30 months. However, long-term follow-up until the eruption of the permanent successor teeth is needed for more definitive assessments.

The influence of methodological variables on the push-out resistance to dislodgement of root filling materials: a meta-regression analysis.

The aim of this systematic review was to evaluate the influence of several methodological variables on the push-out resistance to dislodgment of root filling materials by a meta-regression analysis of the literature. A systematic review was performed by searching the PubMed database using the terms 'push-out' and 'pushout'. Laboratory studies published before March 2015 were included. Two reviewers extracted data regarding country of origin, year of publication, tooth type, smear layer removal, root canal sealer, core material, obturation technique, sample storage, tooth portion, test machine load velocity and slice thickness. Pooled mean resistance to dislodgement of all groups from the included studies was used in a linear meta-regression of random effects (α = 0.05). Of the 850 identified studies, 53 met the inclusion criteria. A meta-regression of the 341 groups extracted from these articles was performed to analyse the influence of each variable on resistance to dislodgement (in MPa). The sealer, core material, obturation technique, slice thickness, storage time, load velocity and tooth portion significantly influence the results (P < 0.05). Irrigant solution and smear layer removal did not influence the resistance to dislodgement of the root filling materials (P > 0.05). Methodological variables such as sealer, core material, root filling technique, tooth type, tooth portion, slice thickness, storage time and load velocity influenced the resistance to dislodgment. The inclusion and standardization of all related variables could lead to a more comparable and reproducible analysis of the resistance to dislodgment of the root canal sealers.

Genetic variability analysis of Byrsonima crassifolia germplasm collected in Pará State using ISSR markers.

Native of the Amazon, the nanche (Byrsonima crassifolia) is a fruit cultivated by family farmers and used in cooking; as such, it represents an opportunity for regional agribusiness. The Embrapa Eastern Amazon set up an active germplasm bank (BAG) consisting of 22 accessions sampled in 11 municipalities of Pará State. Due to its economic potential, there is an interest to advance the genetic breeding program of this species. The aim of this study was to characterize the BAG nanche collection using inter-simple sequence repeat (ISSR) markers. Accessions were genotyped using 23 pre-selected ISSR primers resulting in 109 amplified polymorphic and 51 monomorphic bands. With eight polymorphic bands each, the most polymorphic primers were UBC 809 and UBC 848. An unweighted pair-group method with arithmetic average cluster analysis based on Jaccard's coefficient indicated that the individuals clustered into two distinct groups. Accessions Igarapé Açu-2 and Augusto Corrêa-Pl 1 were most similar. The genetic dissimilarity values ranged from 0.10 to 0.59. We conclude that the ISSR markers were efficient in detecting polymorphisms in the nanche accessions, and that it is possible to infer the genetic variability among accessions of the collection. This demonstrate the importance of using molecular markers in poorly studied species and the advantages that this information can bring to the genetic improvement of such species.